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pubmed23n0125_13577 | Acceptance of hepatitis B vaccine among hospital workers: a follow-up. | We reported earlier on 1,500 hospital employees whose desire for hepatitis B vaccine was assessed. We followed 229 of these subjects to measure the effect of a voluntary educational intervention on desire for vaccine. Acceptance of vaccine declined in 37 per cent of subjects and increased in 13 per cent. One of the extraneous influences on demand for vaccine may have been high-profile media coverage of the AIDS (acquired immune deficiency syndrome) epidemic. | Acceptance of hepatitis B vaccine among hospital workers: a follow-up. We reported earlier on 1,500 hospital employees whose desire for hepatitis B vaccine was assessed. We followed 229 of these subjects to measure the effect of a voluntary educational intervention on desire for vaccine. Acceptance of vaccine declined in 37 per cent of subjects and increased in 13 per cent. One of the extraneous influences on demand for vaccine may have been high-profile media coverage of the AIDS (acquired immune deficiency syndrome) epidemic. | 3,766,834 |
pubmed23n0125_13578 | Hospitalized burn injuries in Massachusetts: an assessment of incidence and product involvement. | We assessed the frequency of hospitalized burn injuries in Massachusetts, and product involvement in causing burns, by reviewing the hospital inpatient records and emergency room logbooks for 240 of New England's 256 acute-care hospitals. Children less than two years of age, males, and Blacks experienced higher burns rates than did older individuals, females, or Whites. Products frequently associated with burn injuries included those involved in food preparation and consumption, flammable liquids, and clothing. | Hospitalized burn injuries in Massachusetts: an assessment of incidence and product involvement. We assessed the frequency of hospitalized burn injuries in Massachusetts, and product involvement in causing burns, by reviewing the hospital inpatient records and emergency room logbooks for 240 of New England's 256 acute-care hospitals. Children less than two years of age, males, and Blacks experienced higher burns rates than did older individuals, females, or Whites. Products frequently associated with burn injuries included those involved in food preparation and consumption, flammable liquids, and clothing. | 3,766,835 |
pubmed23n0125_13579 | The delivery and uptake of nicotine from an aerosol rod. | Nicotine aerosol rods were assessed for their possible usefulness as substitutes for cigarettes. Under standard FTC conditions, the per puff delivery of the aerosol rod averaged 0.3 micrograms nicotine/puff after 10 puffs and 6.4 micrograms nicotine/puff after 60 puffs. After puffing on the rods, no nicotine was detected in the plasma or urine of seven subjects. | The delivery and uptake of nicotine from an aerosol rod. Nicotine aerosol rods were assessed for their possible usefulness as substitutes for cigarettes. Under standard FTC conditions, the per puff delivery of the aerosol rod averaged 0.3 micrograms nicotine/puff after 10 puffs and 6.4 micrograms nicotine/puff after 60 puffs. After puffing on the rods, no nicotine was detected in the plasma or urine of seven subjects. | 3,766,836 |
pubmed23n0125_13580 | The relationship of family income to the incidence, external causes, and outcomes of serious brain injury, San Diego County, California. | Among residents of San Diego County, California the incidence and external causes of serious brain injury were related to the median family income of the census tract of residency. Low income tracts had high incidence rates--a finding not changed by adjustment for age and race/ethnicity. For those injured, the type of emergency transport, time from injury to treatment, and outcome of treatment were not related to the median income of the census tract of residency. | The relationship of family income to the incidence, external causes, and outcomes of serious brain injury, San Diego County, California. Among residents of San Diego County, California the incidence and external causes of serious brain injury were related to the median family income of the census tract of residency. Low income tracts had high incidence rates--a finding not changed by adjustment for age and race/ethnicity. For those injured, the type of emergency transport, time from injury to treatment, and outcome of treatment were not related to the median income of the census tract of residency. | 3,766,837 |
pubmed23n0125_13581 | Impact and cost of public health nurse telephone follow-up of school dental referrals. | The effect and cost of nursing follow-up services on school dental screening outcomes were investigated. Experimental and control groups were randomly assigned. A positive difference in dental visit rate occurred for all nurses, with the overall dentist visit-no visit, experimental-control odds ratio being 1.64 (95% CL = 1.15, 2.35). The service averaged 27.7 minutes and $8.92 per family contacted, suggesting that nursing follow-up increased dental care utilization after screening at low cost in this population. | Impact and cost of public health nurse telephone follow-up of school dental referrals. The effect and cost of nursing follow-up services on school dental screening outcomes were investigated. Experimental and control groups were randomly assigned. A positive difference in dental visit rate occurred for all nurses, with the overall dentist visit-no visit, experimental-control odds ratio being 1.64 (95% CL = 1.15, 2.35). The service averaged 27.7 minutes and $8.92 per family contacted, suggesting that nursing follow-up increased dental care utilization after screening at low cost in this population. | 3,766,838 |
pubmed23n0125_13582 | Birthweight and gestational age: mothers' estimates compared with state and hospital records. | Birthweight and gestational age estimates made by research subjects were compared with values obtained from birth certificates and hospital records. Ninety-one per cent of mothers' estimates of their infant's birthweight and 88 per cent of their estimates of infant gestational age agreed with the delivery record. When the same mothers estimated their own and their husband's birthweight, only 44 per cent were in agreement with birth certificate values; however, 78 per cent were in agreement with the pounds portion of the weight. Agreement of mothers' estimates of her own and her husband's gestational age with birth certificates was no better than chance. | Birthweight and gestational age: mothers' estimates compared with state and hospital records. Birthweight and gestational age estimates made by research subjects were compared with values obtained from birth certificates and hospital records. Ninety-one per cent of mothers' estimates of their infant's birthweight and 88 per cent of their estimates of infant gestational age agreed with the delivery record. When the same mothers estimated their own and their husband's birthweight, only 44 per cent were in agreement with birth certificate values; however, 78 per cent were in agreement with the pounds portion of the weight. Agreement of mothers' estimates of her own and her husband's gestational age with birth certificates was no better than chance. | 3,766,839 |
pubmed23n0125_13583 | Carbohydrates of the surface of the normal human spermatozoon. | Autoproteolysis of human spermatozoa produces oligopeptides with oligosaccharide chains of the N-glycosidic-linked type that are released from the "surface exposed" parts of glycoproteins. The products eluted in the previous washing of the spermatozoa have the same composition and solubility characteristics as the oligopeptides from the digestion. This suggests that autoproteolysis is a constant process that normally occurs on the spermatozoa membrane. The cytochemical characterization and localization of the N-glycosidic-linked oligosaccharide receptors on the human spermatozoa membrane after digestion, in the presence or absence of seminal plasma, indicates that only part of the oligosaccharides are cleaved. Their distribution on the different zones of the spermatozoon changed as the probability of detecting these receptors in the intermediate segment increased after proteolysis; this indicates that in this zone the receptors are cryptic ones that become exposed by the action of the proteolytic enzymes. In the presence of seminal plasma most receptors on the acrosome are eliminated. | Carbohydrates of the surface of the normal human spermatozoon. Autoproteolysis of human spermatozoa produces oligopeptides with oligosaccharide chains of the N-glycosidic-linked type that are released from the "surface exposed" parts of glycoproteins. The products eluted in the previous washing of the spermatozoa have the same composition and solubility characteristics as the oligopeptides from the digestion. This suggests that autoproteolysis is a constant process that normally occurs on the spermatozoa membrane. The cytochemical characterization and localization of the N-glycosidic-linked oligosaccharide receptors on the human spermatozoa membrane after digestion, in the presence or absence of seminal plasma, indicates that only part of the oligosaccharides are cleaved. Their distribution on the different zones of the spermatozoon changed as the probability of detecting these receptors in the intermediate segment increased after proteolysis; this indicates that in this zone the receptors are cryptic ones that become exposed by the action of the proteolytic enzymes. In the presence of seminal plasma most receptors on the acrosome are eliminated. | 3,766,843 |
pubmed23n0125_13584 | Adoptive transfer of suppression of autoimmune response to rat male accessory glands with spleen mononuclear cells from antigen-pretreated rats. | The injection of the spleen mononuclear (SpM) cells, obtained from rats rendered unresponsive to autoantigen of rat male accessory glands (RAG) by pretreatment with low doses of purified fraction of RAG (containing the autoantigen), into normal syngeneic recipients markedly prevented the development of delayed type hypersensitivity (DTH) reaction to the autoantigen (suppression of the induction) (p less than 0.001). The humoral response was not altered. The control animals were rat receptors of spleen mononuclear cells from donor rats pretreated with rat lung saline extract or 0.15 M NaCl. In contrast, the transference of SpM cells from donor rats pretreated with low doses of autoantigen prior to the immunization to rats previously immunized, did not modify the expression of the immune response against the autoantigen when compared with control rats. The suppression of the induction of DTH response was also obtained when prior to the immunization, the recipients received T-cell-enriched SpM cell population from unresponsive animals (p less than 0.001), but not when they were injected with B-cell-enriched SpM cells. These results suggest that suppressor T cells capable of controlling induction of the autoimmune response against RAG autoantigen might be one of the immunoregulatory mechanisms that are activated when soluble autoantigen of RAG enter into circulation. | Adoptive transfer of suppression of autoimmune response to rat male accessory glands with spleen mononuclear cells from antigen-pretreated rats. The injection of the spleen mononuclear (SpM) cells, obtained from rats rendered unresponsive to autoantigen of rat male accessory glands (RAG) by pretreatment with low doses of purified fraction of RAG (containing the autoantigen), into normal syngeneic recipients markedly prevented the development of delayed type hypersensitivity (DTH) reaction to the autoantigen (suppression of the induction) (p less than 0.001). The humoral response was not altered. The control animals were rat receptors of spleen mononuclear cells from donor rats pretreated with rat lung saline extract or 0.15 M NaCl. In contrast, the transference of SpM cells from donor rats pretreated with low doses of autoantigen prior to the immunization to rats previously immunized, did not modify the expression of the immune response against the autoantigen when compared with control rats. The suppression of the induction of DTH response was also obtained when prior to the immunization, the recipients received T-cell-enriched SpM cell population from unresponsive animals (p less than 0.001), but not when they were injected with B-cell-enriched SpM cells. These results suggest that suppressor T cells capable of controlling induction of the autoimmune response against RAG autoantigen might be one of the immunoregulatory mechanisms that are activated when soluble autoantigen of RAG enter into circulation. | 3,766,844 |
pubmed23n0125_13585 | Clear cell cribriform hyperplasia of prostate. Report of 10 cases. | We report 10 patients with clear cell cribriform hyperplasia of the prostate. Their ages ranged from 62 to 87 years, with a mean of 72 years. The clinical diagnosis in all patients was benign nodular hyperplasia; all the patients are alive and have shown no evidence of recurrent disease. Follow-ups ranged from 1 month to 7 years (median: 12.5 months; mean: 24.6 months). Pathologically, this lesion has a cribriform arrangement of clear cells with a complex papillary growth simulating the cribriform pattern of prostatic carcinoma. In fact, in five of the 10 cases, the referring diagnosis was either carcinoma or possible carcinoma. Cytologically, however, there is no nuclear atypia, mitosis, or prominent nucleoli, and typically there is a double epithelial cell layer at the periphery of the involved acini. In summary, clear cell cribriform hyperplasia is a benign hyperplastic process with a complex papillary-cribriform structure and should not be confused with prostatic carcinoma. The key feature for the diagnosis is the preservation of nodular configuration with a bland cytology and double cell layer lining the involved acini. | Clear cell cribriform hyperplasia of prostate. Report of 10 cases. We report 10 patients with clear cell cribriform hyperplasia of the prostate. Their ages ranged from 62 to 87 years, with a mean of 72 years. The clinical diagnosis in all patients was benign nodular hyperplasia; all the patients are alive and have shown no evidence of recurrent disease. Follow-ups ranged from 1 month to 7 years (median: 12.5 months; mean: 24.6 months). Pathologically, this lesion has a cribriform arrangement of clear cells with a complex papillary growth simulating the cribriform pattern of prostatic carcinoma. In fact, in five of the 10 cases, the referring diagnosis was either carcinoma or possible carcinoma. Cytologically, however, there is no nuclear atypia, mitosis, or prominent nucleoli, and typically there is a double epithelial cell layer at the periphery of the involved acini. In summary, clear cell cribriform hyperplasia is a benign hyperplastic process with a complex papillary-cribriform structure and should not be confused with prostatic carcinoma. The key feature for the diagnosis is the preservation of nodular configuration with a bland cytology and double cell layer lining the involved acini. | 3,766,845 |
pubmed23n0125_13586 | The grooved nucleus. A useful diagnostic criterion of papillary carcinoma of the thyroid. | The optically clear nucleus has been recognized widely as an important histological feature of papillary carcinoma of the thyroid, and it is now considered by many workers as standing on an equal basis with the papillae for the diagnosis of this tumor. However, clear nuclei are sometimes patchy in distribution or absent in papillary carcinoma, and may be difficult to differentiate from pseudoclear nuclei which may be found in a variety of thyroid lesions. This study was undertaken to determine whether the grooving of the nuclei, a well-recognized electron-microscopic feature, can serve as an additional reliable light microscopic diagnostic criterion of papillary carcinoma. The histological sections of 103 consecutive cases of thyroid carcinoma, (including 89 cases of papillary carcinoma), 100 consecutive noncancer thyroidectomy specimens, and five cases of Hashimoto's thyroiditis were reviewed. Grooved nuclei were found in all cases of papillary carcinoma in almost every high-power microscopic field, and in the two cases of poorly differentiated thyroid carcinoma. Similar nuclei were found in only a very occasional field in four cases of adenoma, and one case each of multinodular goiter and Hashimoto's thyroiditis. We conclude that the deep grooving of the nuclei is a useful cytological feature for the diagnosis of papillary thyroid carcinoma. | The grooved nucleus. A useful diagnostic criterion of papillary carcinoma of the thyroid. The optically clear nucleus has been recognized widely as an important histological feature of papillary carcinoma of the thyroid, and it is now considered by many workers as standing on an equal basis with the papillae for the diagnosis of this tumor. However, clear nuclei are sometimes patchy in distribution or absent in papillary carcinoma, and may be difficult to differentiate from pseudoclear nuclei which may be found in a variety of thyroid lesions. This study was undertaken to determine whether the grooving of the nuclei, a well-recognized electron-microscopic feature, can serve as an additional reliable light microscopic diagnostic criterion of papillary carcinoma. The histological sections of 103 consecutive cases of thyroid carcinoma, (including 89 cases of papillary carcinoma), 100 consecutive noncancer thyroidectomy specimens, and five cases of Hashimoto's thyroiditis were reviewed. Grooved nuclei were found in all cases of papillary carcinoma in almost every high-power microscopic field, and in the two cases of poorly differentiated thyroid carcinoma. Similar nuclei were found in only a very occasional field in four cases of adenoma, and one case each of multinodular goiter and Hashimoto's thyroiditis. We conclude that the deep grooving of the nuclei is a useful cytological feature for the diagnosis of papillary thyroid carcinoma. | 3,766,846 |
pubmed23n0125_13587 | Giant cell arteritis of the female genital tract. A report of three cases. | Three cases of giant cell arteritis involving the female genital tract of postmenopausal women are reported. The patients were 80, 64, and 57 years of age and presented with fatigue and anemia, fatigue and an abdominal mass, and fever and weight loss, respectively. Two of the patients had palpable pelvic masses; one had an ovarian mass visible on ultrasound examination. All three patients were anemic, and the erythrocyte sedimentation rate was elevated in the two women in whom it was tested. Exploratory laparotomy revealed ovarian tumors in two patients; one had a mucinous cystadenoma, and one had bilateral ovarian fibromas. The third patient had a cyst of the rete ovarii. Extensive giant cell arteritis of the small to medium-sized arteries was found unexpectedly in the ovaries and fallopian tubes of two patients who had prior hysterectomies and in the ovaries, fallopian tubes, and uterus of one patient. One patient was treated postoperatively with prednisone with improvement of symptoms and a decrease in the erythrocyte sedimentation rate. Of the two patients who received no therapy, one was found to have a thoracic aortic aneurysm 5 years postoperatively, and the other was alive without symptoms 17 years after the operation. Giant cell arteritis of the female genital tract is a rare finding in elderly women and may occur as an isolated finding or as part of generalized giant cell arteritis. | Giant cell arteritis of the female genital tract. A report of three cases. Three cases of giant cell arteritis involving the female genital tract of postmenopausal women are reported. The patients were 80, 64, and 57 years of age and presented with fatigue and anemia, fatigue and an abdominal mass, and fever and weight loss, respectively. Two of the patients had palpable pelvic masses; one had an ovarian mass visible on ultrasound examination. All three patients were anemic, and the erythrocyte sedimentation rate was elevated in the two women in whom it was tested. Exploratory laparotomy revealed ovarian tumors in two patients; one had a mucinous cystadenoma, and one had bilateral ovarian fibromas. The third patient had a cyst of the rete ovarii. Extensive giant cell arteritis of the small to medium-sized arteries was found unexpectedly in the ovaries and fallopian tubes of two patients who had prior hysterectomies and in the ovaries, fallopian tubes, and uterus of one patient. One patient was treated postoperatively with prednisone with improvement of symptoms and a decrease in the erythrocyte sedimentation rate. Of the two patients who received no therapy, one was found to have a thoracic aortic aneurysm 5 years postoperatively, and the other was alive without symptoms 17 years after the operation. Giant cell arteritis of the female genital tract is a rare finding in elderly women and may occur as an isolated finding or as part of generalized giant cell arteritis. | 3,766,847 |
pubmed23n0125_13588 | Pelvic lymph node metastases in prostatic carcinoma. The value of touch imprint cytology. | Touch imprint cytology is a simple, rapid, reliable, and highly sensitive procedure when used for detecting lymph node metastases in men with clinically localized prostatic adenocarcinoma who are undergoing staging pelvic lymphadenectomy. Thirty-five men admitted for this procedure had frozen sections performed on the lymph nodes removed at surgery. Of these 35 patients, 19 had touch imprints made of their lymph nodes. In five patients classified as having Stage D disease, positive frozen section correlated with permanent section. In two of these cases, touch imprint cytology was performed and was positive for malignant cells. In seven patients classified as having Stage D disease, frozen sections were negative. Permanent sections were positive for metastatic tumor in five of these patients. In four of these cases, touch imprint cytology was performed. In two cases, cytology and permanent sections were positive and, in two cases, cytology alone was positive. Factors that contributed to the false negative rate for frozen section were: the presence of micrometastases, fatty lymph nodes that were difficult to process, and sampling errors on the part of the pathologist. Touch imprint cytology overcame the deficits of frozen section and proved useful in detecting metastatic disease in lymph nodes. | Pelvic lymph node metastases in prostatic carcinoma. The value of touch imprint cytology. Touch imprint cytology is a simple, rapid, reliable, and highly sensitive procedure when used for detecting lymph node metastases in men with clinically localized prostatic adenocarcinoma who are undergoing staging pelvic lymphadenectomy. Thirty-five men admitted for this procedure had frozen sections performed on the lymph nodes removed at surgery. Of these 35 patients, 19 had touch imprints made of their lymph nodes. In five patients classified as having Stage D disease, positive frozen section correlated with permanent section. In two of these cases, touch imprint cytology was performed and was positive for malignant cells. In seven patients classified as having Stage D disease, frozen sections were negative. Permanent sections were positive for metastatic tumor in five of these patients. In four of these cases, touch imprint cytology was performed. In two cases, cytology and permanent sections were positive and, in two cases, cytology alone was positive. Factors that contributed to the false negative rate for frozen section were: the presence of micrometastases, fatty lymph nodes that were difficult to process, and sampling errors on the part of the pathologist. Touch imprint cytology overcame the deficits of frozen section and proved useful in detecting metastatic disease in lymph nodes. | 3,766,848 |
pubmed23n0125_13589 | The incidence of diarrheal disease in a defined population of rural Egypt. | A prospective study of diarrheal illness was conducted for 2 years in 309 families who were part of an epidemiologic research center population in rural Egypt. The rates of illness peaked in the first year of life with between 5 and 6 episodes per person year at risk. Although no seasonal variation in reported diarrhea was demonstrated for the total population, age-specific differences in seasonal occurrence were noted. The incidence did not consistently vary by village size, nor did rates vary by sex. | The incidence of diarrheal disease in a defined population of rural Egypt. A prospective study of diarrheal illness was conducted for 2 years in 309 families who were part of an epidemiologic research center population in rural Egypt. The rates of illness peaked in the first year of life with between 5 and 6 episodes per person year at risk. Although no seasonal variation in reported diarrhea was demonstrated for the total population, age-specific differences in seasonal occurrence were noted. The incidence did not consistently vary by village size, nor did rates vary by sex. | 3,766,849 |
pubmed23n0125_13590 | Blastocystis hominis: pathogen or fellow traveler? | Blastocystis hominis, an intestinal organism of uncertain taxonomic position, has long been considered nonpathogenic. Some recent studies suggest, however, that it may be associated with diarrhea and may respond to treatment with iodoquinol or metronidazole. To investigate this possibility, we identified 148 persons whose stools contained this organism. Of this number, 32 had at least 6 stool examinations performed. Twenty-seven of the 32 persons were later found to have greater than or equal to 1 recognized pathogens--Entamoeba histolytica, Giardia lamblia or Dientamoeba fragilis--and, after receiving appropriate therapy, became asymptomatic. The B hominis infection, however, was unaffected by therapy. Five persons with only B. hominis infection were treated with iodoquinol without effect; these persons fulfilled the medical criteria for irritable bowel syndrome. We believe that when an apparently symptomatic B. hominis infection responds to therapy, the improvement probably represents elimination of some other undetected organism causing the infection. | Blastocystis hominis: pathogen or fellow traveler? Blastocystis hominis, an intestinal organism of uncertain taxonomic position, has long been considered nonpathogenic. Some recent studies suggest, however, that it may be associated with diarrhea and may respond to treatment with iodoquinol or metronidazole. To investigate this possibility, we identified 148 persons whose stools contained this organism. Of this number, 32 had at least 6 stool examinations performed. Twenty-seven of the 32 persons were later found to have greater than or equal to 1 recognized pathogens--Entamoeba histolytica, Giardia lamblia or Dientamoeba fragilis--and, after receiving appropriate therapy, became asymptomatic. The B hominis infection, however, was unaffected by therapy. Five persons with only B. hominis infection were treated with iodoquinol without effect; these persons fulfilled the medical criteria for irritable bowel syndrome. We believe that when an apparently symptomatic B. hominis infection responds to therapy, the improvement probably represents elimination of some other undetected organism causing the infection. | 3,766,850 |
pubmed23n0125_13591 | Etiologic and serologic investigations of the 1980 epidemic of dengue fever on Hainan Island, China. | Virologic and seroepidemiologic studies were carried out during an epidemic of dengue fever on Hainan Island in 1980. Dengue 3 virus was isolated from 46 of 77 acute phase sera and from 1 of 10 pools of adult Aedes aegypti. Dengue 1 virus virus was isolated from a single acute phase serum. Seroepidemiologic investigations showed that 74% of healthy individuals in the epidemic area had antibody to dengue virus compared to 54% in an area where epidemic dengue had occurred in 1978, and less than or equal to 8% in nonepidemic areas. There were no significant differences in antibody prevalence for different sex and age groups. | Etiologic and serologic investigations of the 1980 epidemic of dengue fever on Hainan Island, China. Virologic and seroepidemiologic studies were carried out during an epidemic of dengue fever on Hainan Island in 1980. Dengue 3 virus was isolated from 46 of 77 acute phase sera and from 1 of 10 pools of adult Aedes aegypti. Dengue 1 virus virus was isolated from a single acute phase serum. Seroepidemiologic investigations showed that 74% of healthy individuals in the epidemic area had antibody to dengue virus compared to 54% in an area where epidemic dengue had occurred in 1978, and less than or equal to 8% in nonepidemic areas. There were no significant differences in antibody prevalence for different sex and age groups. | 3,766,851 |
pubmed23n0125_13592 | Rift Valley fever surveillance in mobile sheep flocks in the Nile Delta. | Rift Valley fever (RVF) surveillance was carried out in the Nile Delta by monitoring mobile and stationary sheep flocks for antibodies to RVF virus. Sheep are known to be susceptible to RVF virus infection and experienced severe morbidity in 1977 and 1978 when RVF was epidemic in Egypt. Four hundred six sheep in 32 flocks were surveyed during 1984. Twenty-four sheep from 7 flocks had antibodies to RVF virus detected by hemagglutination inhibition and plaque reduction neutralization tests. Antibodies were found primarily in sheep greater than 3 years of age, although 1- and 2-year-old sheep were included in the sample. No seroconversion was observed among 177 seronegative sheep that were bled successively for a period of 10 months. These results indicate that epizootic RVF was probably not present in the Nile Delta during 1984. | Rift Valley fever surveillance in mobile sheep flocks in the Nile Delta. Rift Valley fever (RVF) surveillance was carried out in the Nile Delta by monitoring mobile and stationary sheep flocks for antibodies to RVF virus. Sheep are known to be susceptible to RVF virus infection and experienced severe morbidity in 1977 and 1978 when RVF was epidemic in Egypt. Four hundred six sheep in 32 flocks were surveyed during 1984. Twenty-four sheep from 7 flocks had antibodies to RVF virus detected by hemagglutination inhibition and plaque reduction neutralization tests. Antibodies were found primarily in sheep greater than 3 years of age, although 1- and 2-year-old sheep were included in the sample. No seroconversion was observed among 177 seronegative sheep that were bled successively for a period of 10 months. These results indicate that epizootic RVF was probably not present in the Nile Delta during 1984. | 3,766,852 |
pubmed23n0125_13593 | The effect of post-bloodmeal nutrition of Phlebotomus papatasi on the transmission of Leishmania major. | To test the effects of post-bloodmeal nutrition of sand flies on the transmission of Leishmania major, groups of infected P. papatasi females maintained on diets of sucrose, trehalose, albumin or a mixture of sucrose and albumin, were subjected to forced feeding with capillaries. Transmission was evaluated by counting the parasites egested; numbers ranged from 0 to over 1,000 promastigotes. Infections of the anterior midgut were seen in the majority of flies from all the experimental groups but the percentage of transmitting females was significantly higher in the group maintained on a mixture of sucrose and albumin. There were no attached parasites in the pharynx and cibarium of the flies and the presence of free promastigotes in these parts was not itself indicative of infectivity. However, transmission was positively correlated with apparent inability to engorge. The parasites egested were typical infective form promastigotes and identical to those observed in the esophagus and the anterior thoracic midgut. A mechanism by which infective stage promastigotes from the esophagus and the stomodeal valve may be transmitted by bite is proposed. | The effect of post-bloodmeal nutrition of Phlebotomus papatasi on the transmission of Leishmania major. To test the effects of post-bloodmeal nutrition of sand flies on the transmission of Leishmania major, groups of infected P. papatasi females maintained on diets of sucrose, trehalose, albumin or a mixture of sucrose and albumin, were subjected to forced feeding with capillaries. Transmission was evaluated by counting the parasites egested; numbers ranged from 0 to over 1,000 promastigotes. Infections of the anterior midgut were seen in the majority of flies from all the experimental groups but the percentage of transmitting females was significantly higher in the group maintained on a mixture of sucrose and albumin. There were no attached parasites in the pharynx and cibarium of the flies and the presence of free promastigotes in these parts was not itself indicative of infectivity. However, transmission was positively correlated with apparent inability to engorge. The parasites egested were typical infective form promastigotes and identical to those observed in the esophagus and the anterior thoracic midgut. A mechanism by which infective stage promastigotes from the esophagus and the stomodeal valve may be transmitted by bite is proposed. | 3,766,853 |
pubmed23n0125_13594 | Clinical evaluation of amoscanate in healthy male volunteers. | Single oral doses of amoscanate (4-isothiocyanato-4'-nitrodiphenylamine), an experimental antiparasitic agent, are highly effective in animals against the four major species of schistosomes which infect humans. Two prospective, randomized, double blinded, placebo controlled Phase I studies were designed to evaluate the tolerance and safety of the 5% aqueous suspension of 2-mu particles of amoscanate administered to healthy male volunteers. In addition to routine safety monitoring, particular attention was directed toward detecting hepatic, neurological, cardiovascular or ocular toxicity. Three of four men who received 3.5 mg/kg of amoscanate developed mild, reversible hepatotoxicity, which could not be unequivocably attributed to the drug. In the second study, of 1 mg/kg amoscanate, there was no statistically significant evidence of hepatotoxicity, although 1 of 12 drug recipients developed transient liver chemistry changes. Despite intensive monitoring, there was no evidence in either study of significant symptomatic complaints, or of neurological, cardiovascular or ocular toxicity. No mutagenic activity attributable to amoscanate was detectable in the urine. These results suggest that this formulation of amoscanate, at 1 mg/kg, is sufficiently well tolerated and safe to justify evaluation for efficacy in patients with schistosomiasis. | Clinical evaluation of amoscanate in healthy male volunteers. Single oral doses of amoscanate (4-isothiocyanato-4'-nitrodiphenylamine), an experimental antiparasitic agent, are highly effective in animals against the four major species of schistosomes which infect humans. Two prospective, randomized, double blinded, placebo controlled Phase I studies were designed to evaluate the tolerance and safety of the 5% aqueous suspension of 2-mu particles of amoscanate administered to healthy male volunteers. In addition to routine safety monitoring, particular attention was directed toward detecting hepatic, neurological, cardiovascular or ocular toxicity. Three of four men who received 3.5 mg/kg of amoscanate developed mild, reversible hepatotoxicity, which could not be unequivocably attributed to the drug. In the second study, of 1 mg/kg amoscanate, there was no statistically significant evidence of hepatotoxicity, although 1 of 12 drug recipients developed transient liver chemistry changes. Despite intensive monitoring, there was no evidence in either study of significant symptomatic complaints, or of neurological, cardiovascular or ocular toxicity. No mutagenic activity attributable to amoscanate was detectable in the urine. These results suggest that this formulation of amoscanate, at 1 mg/kg, is sufficiently well tolerated and safe to justify evaluation for efficacy in patients with schistosomiasis. | 3,766,854 |
pubmed23n0125_13595 | Reliable serology of Taenia solium cysticercosis with antigens from cyst vesicular fluid: ELISA and hemagglutination tests. | New levels of reproducibility and sensitivity have been achieved in the detection of anticysticercus antibodies in human sera by using cysticercus vesicular fluid as the source of antigens for both ELISA and hemagglutination assays. Reproducibility both between tests on a serum and between similar sera was significantly improved over typical results using antigens from whole parasite extracts. Sera collected from uninfected individuals in endemic areas gave somewhat elevated values over those collected in nonendemic areas. This necessitated the use of a higher threshold in endemic areas to avoid false positives. With the threshold appropriate for a nonendemic area, both ELISA and hemagglutination were sensitive enough to detect infection in 95% of cases. With the threshold value for sera from an endemic area, these sensitivities were reduced to 80%-90%. A prominent 103-Kd protein of vesicular fluid, not related to antigen B, elicited the strongest antibody response in neurocystercotic patients. | Reliable serology of Taenia solium cysticercosis with antigens from cyst vesicular fluid: ELISA and hemagglutination tests. New levels of reproducibility and sensitivity have been achieved in the detection of anticysticercus antibodies in human sera by using cysticercus vesicular fluid as the source of antigens for both ELISA and hemagglutination assays. Reproducibility both between tests on a serum and between similar sera was significantly improved over typical results using antigens from whole parasite extracts. Sera collected from uninfected individuals in endemic areas gave somewhat elevated values over those collected in nonendemic areas. This necessitated the use of a higher threshold in endemic areas to avoid false positives. With the threshold appropriate for a nonendemic area, both ELISA and hemagglutination were sensitive enough to detect infection in 95% of cases. With the threshold value for sera from an endemic area, these sensitivities were reduced to 80%-90%. A prominent 103-Kd protein of vesicular fluid, not related to antigen B, elicited the strongest antibody response in neurocystercotic patients. | 3,766,855 |
pubmed23n0125_13596 | Ultrastructural observations on the fate of Brugia malayi in jirds previously vaccinated with irradiated infective stage larvae. | Vaccination of inbred jirds (Meriones unguiculatus) with 60cobalt radiation-attenuated Brugia malayi infective stage larvae (L3) protected against homologous challenge given either subcutaneously (sc) or by the intraperitoneal route (ip). At necropsy numerous nodules were recovered from the peritoneal cavities of jirds which had been vaccinated sc and challenged ip. Histopathologic analysis showed these to be granulomas containing dead and dying larvae and transmission electron microscopy showed that eosinophils were present in high numbers around and within the larvae. Structural damage to the L3 cuticle was apparent in discrete areas and eosinophils actively entering the breached cuticle at the time of fixation were observed. Coalescence of eosinophil secretion granules and the formation of degranulation vacuoles were seen in eosinophils throughout the granulomas. Degranulation resulted in the deposition of electron-dense material on the surface of the larval cuticle. The jird vaccine model for B. malayi thus appears to be a potentially useful tool for investigation of immune mechanisms in filariasis. | Ultrastructural observations on the fate of Brugia malayi in jirds previously vaccinated with irradiated infective stage larvae. Vaccination of inbred jirds (Meriones unguiculatus) with 60cobalt radiation-attenuated Brugia malayi infective stage larvae (L3) protected against homologous challenge given either subcutaneously (sc) or by the intraperitoneal route (ip). At necropsy numerous nodules were recovered from the peritoneal cavities of jirds which had been vaccinated sc and challenged ip. Histopathologic analysis showed these to be granulomas containing dead and dying larvae and transmission electron microscopy showed that eosinophils were present in high numbers around and within the larvae. Structural damage to the L3 cuticle was apparent in discrete areas and eosinophils actively entering the breached cuticle at the time of fixation were observed. Coalescence of eosinophil secretion granules and the formation of degranulation vacuoles were seen in eosinophils throughout the granulomas. Degranulation resulted in the deposition of electron-dense material on the surface of the larval cuticle. The jird vaccine model for B. malayi thus appears to be a potentially useful tool for investigation of immune mechanisms in filariasis. | 3,766,856 |
pubmed23n0125_13597 | Dirofilaria from the index finger of a man in Costa Rica. | A 4.5-cm male Dirofilaria indistinguishable from D. immitis was removed from the tip of the index finger of an 18-year-old farmer in Costa Rica. The case is reminiscent of a report of a larger (9-cm) but otherwise similar Dirofilaria, identified as D. spectans, a heartworm of otters, causing Raynaud's syndrome by occluding an artery of the fifth finger of a farm woman in Brazil. As in the present case, the worm was removed from a painful papule on the volar surface at the extreme tip of the affected finger. | Dirofilaria from the index finger of a man in Costa Rica. A 4.5-cm male Dirofilaria indistinguishable from D. immitis was removed from the tip of the index finger of an 18-year-old farmer in Costa Rica. The case is reminiscent of a report of a larger (9-cm) but otherwise similar Dirofilaria, identified as D. spectans, a heartworm of otters, causing Raynaud's syndrome by occluding an artery of the fifth finger of a farm woman in Brazil. As in the present case, the worm was removed from a painful papule on the volar surface at the extreme tip of the affected finger. | 3,766,857 |
pubmed23n0125_13598 | Orbital hydatid disease in Kenya. | We treated 7 cases of orbital hydatidosis presenting as cysts. These were successfully treated surgically using a technique that does not compromise the eye and its visual function. This need arose when it was learned that 7 eyes had been enucleated and another case put on massive steroid therapy due to misdiagnosis as malignancies by ophthalmologists visiting in this country (M. French, African Medical Research Foundation, personal communication). It is imperative to stress the diagnostic and therapeutic aspects of this condition which must be considered in the differential diagnosis of unilateral proptosis. | Orbital hydatid disease in Kenya. We treated 7 cases of orbital hydatidosis presenting as cysts. These were successfully treated surgically using a technique that does not compromise the eye and its visual function. This need arose when it was learned that 7 eyes had been enucleated and another case put on massive steroid therapy due to misdiagnosis as malignancies by ophthalmologists visiting in this country (M. French, African Medical Research Foundation, personal communication). It is imperative to stress the diagnostic and therapeutic aspects of this condition which must be considered in the differential diagnosis of unilateral proptosis. | 3,766,858 |
pubmed23n0125_13599 | Predictive value of tumor thickness in squamous carcinoma confined to the tongue and floor of the mouth. | In this review of 105 consecutive patients who underwent operation for previously untreated, N0 squamous carcinomas arising in the oral tongue or the floor of the mouth, 86 percent of the determinate patients remained alive and well 2 years after treatment. Included were 48 patients, 49 patients, and 8 patients who had T1, T2, and T3 tumors respectively. Elective cervical lymphadenectomy was performed in about a third, but tumor staging did not facilitate selection of those who were most likely to have occult metastases. For this reason, we retrospectively assessed the impact of tumor thickness using an optical micrometer to measure the thickness in millimeters of the excised tumors in routinely prepared paraffin sections. Disease-related death appears to be unusual when oral tumors are thin (2 mm or less), regardless of the tumor stage. Multivariate analysis confirms that increasing tumor thickness, rather than tumor stage, had the best correlation with treatment failure and survival. These findings need to be verified in prospective studies involving a larger patient population and other head and neck sites, but they strongly suggest that measurement of tumor thickness may be a better way to select those oral cancer patients who are most likely to benefit from elective treatment of the N0 neck. | Predictive value of tumor thickness in squamous carcinoma confined to the tongue and floor of the mouth. In this review of 105 consecutive patients who underwent operation for previously untreated, N0 squamous carcinomas arising in the oral tongue or the floor of the mouth, 86 percent of the determinate patients remained alive and well 2 years after treatment. Included were 48 patients, 49 patients, and 8 patients who had T1, T2, and T3 tumors respectively. Elective cervical lymphadenectomy was performed in about a third, but tumor staging did not facilitate selection of those who were most likely to have occult metastases. For this reason, we retrospectively assessed the impact of tumor thickness using an optical micrometer to measure the thickness in millimeters of the excised tumors in routinely prepared paraffin sections. Disease-related death appears to be unusual when oral tumors are thin (2 mm or less), regardless of the tumor stage. Multivariate analysis confirms that increasing tumor thickness, rather than tumor stage, had the best correlation with treatment failure and survival. These findings need to be verified in prospective studies involving a larger patient population and other head and neck sites, but they strongly suggest that measurement of tumor thickness may be a better way to select those oral cancer patients who are most likely to benefit from elective treatment of the N0 neck. | 3,766,861 |
pubmed23n0125_13600 | Relation of thickness of floor of mouth stage I and II cancers to regional metastasis. | We have reviewed the clinical course and histologic findings in 84 patients with stage I and II squamous carcinoma of the mouth floor. We concluded that lesion thickness may offer a useful method for predicting the probability of cervical metastasis in node negative (N0) patients. Moreover, surface area of the lesion did not correlate with subsequent nodal disease, whereas thickness did. Elective node dissection appears to be strongly indicated in any patient with a N0 lesion measuring more than 1.5 mm in thickness. | Relation of thickness of floor of mouth stage I and II cancers to regional metastasis. We have reviewed the clinical course and histologic findings in 84 patients with stage I and II squamous carcinoma of the mouth floor. We concluded that lesion thickness may offer a useful method for predicting the probability of cervical metastasis in node negative (N0) patients. Moreover, surface area of the lesion did not correlate with subsequent nodal disease, whereas thickness did. Elective node dissection appears to be strongly indicated in any patient with a N0 lesion measuring more than 1.5 mm in thickness. | 3,766,862 |
pubmed23n0125_13601 | Microscopic cut-through of cancer in the surgical treatment of squamous carcinoma of the tongue. Prognostic and therapeutic implications. | A retrospective review was carried out of the records of 268 patients with a pathologically proved diagnosis of squamous carcinoma of the tongue treated by glossectomy at M.D. Anderson Hospital and Tumor Institute from January 1, 1970 through December 31, 1979. Pathologic findings found on frozen section analysis were correlated with the TNM stage of the tumor, histologic characteristics, perineural invasion, type of treatment, local recurrence, and survival rate. Thirty-eight patients with squamous carcinoma of the oral tongue and 16 patients with tumors of the base of the tongue had initially positive frozen section margins. Forty-one of these patients had margins that were negative at the completion of surgery. Thirteen patients never had negative margins. Positive mucosal margins were more common in smaller tumors and positive muscular margins were more common in larger tumors. Patients with perineural invasion and larger tumors had a higher percentage of positive margins. Those patients with initially positive margins that were rendered negative at the completion of the procedure and treated with surgery only had a significantly increased local recurrence rate and reduced survival compared with patients similarly treated with initially negative margins. Tumor stage, location (muscular versus mucosal) of positive margins, and the presence of perineural invasion were not significant in predicting local recurrence or decreased survival. If all patients with microscopically positive margins are considered for postoperative radiotherapy, then the routine use of intraoperative frozen section examination of the margins of resection in squamous carcinoma of the tongue may not be justified. | Microscopic cut-through of cancer in the surgical treatment of squamous carcinoma of the tongue. Prognostic and therapeutic implications. A retrospective review was carried out of the records of 268 patients with a pathologically proved diagnosis of squamous carcinoma of the tongue treated by glossectomy at M.D. Anderson Hospital and Tumor Institute from January 1, 1970 through December 31, 1979. Pathologic findings found on frozen section analysis were correlated with the TNM stage of the tumor, histologic characteristics, perineural invasion, type of treatment, local recurrence, and survival rate. Thirty-eight patients with squamous carcinoma of the oral tongue and 16 patients with tumors of the base of the tongue had initially positive frozen section margins. Forty-one of these patients had margins that were negative at the completion of surgery. Thirteen patients never had negative margins. Positive mucosal margins were more common in smaller tumors and positive muscular margins were more common in larger tumors. Patients with perineural invasion and larger tumors had a higher percentage of positive margins. Those patients with initially positive margins that were rendered negative at the completion of the procedure and treated with surgery only had a significantly increased local recurrence rate and reduced survival compared with patients similarly treated with initially negative margins. Tumor stage, location (muscular versus mucosal) of positive margins, and the presence of perineural invasion were not significant in predicting local recurrence or decreased survival. If all patients with microscopically positive margins are considered for postoperative radiotherapy, then the routine use of intraoperative frozen section examination of the margins of resection in squamous carcinoma of the tongue may not be justified. | 3,766,863 |
pubmed23n0125_13602 | Cervical lymph node metastasis after local excision of early squamous cell carcinoma of the oral cavity. | A total of 54 patients with stage I and stage II squamous cell carcinoma of the oral cavity were reviewed as to treatment modality, adequacy of treatment, and site of failure. Surgery was employed as the sole initial treatment modality in 52 patients. Forty-three underwent primary tumor excision alone and 9 underwent elective neck dissection at the time of primary tumor excision. The patients who underwent elective neck dissection at the time of excision of the primary tumor had a 3 year survival rate of 88 percent, in comparison to a survival rate of 77 percent in those patients whose initial therapy was directed solely at the primary tumor. A low incidence of local recurrence (2 percent) and a high incidence of neck recurrence (42 percent) were documented in those patients treated by primary tumor excision alone. Patients who underwent salvage neck dissection for recurrent neck node metastases had a 3 year survival rate of 56 percent. This study has documented a high incidence of cervical node recurrence in patients with T1 and T2 squamous cell carcinomas of the oral cavity treated by primary tumor excision alone and a poor survival rate after salvage therapy. A small group of patients who underwent elective neck dissection had a demonstrably high survival rate. These observations lend support to the call for elective neck dissection in patients with stage I and II oral cavity carcinoma but are not conclusive. Therapeutic decisions regarding elective treatment of the neck will continue to be made according to the best judgment and prejudices of the individual surgeon until a prospective, randomized multi-institutional study addressing this specific issue is undertaken. | Cervical lymph node metastasis after local excision of early squamous cell carcinoma of the oral cavity. A total of 54 patients with stage I and stage II squamous cell carcinoma of the oral cavity were reviewed as to treatment modality, adequacy of treatment, and site of failure. Surgery was employed as the sole initial treatment modality in 52 patients. Forty-three underwent primary tumor excision alone and 9 underwent elective neck dissection at the time of primary tumor excision. The patients who underwent elective neck dissection at the time of excision of the primary tumor had a 3 year survival rate of 88 percent, in comparison to a survival rate of 77 percent in those patients whose initial therapy was directed solely at the primary tumor. A low incidence of local recurrence (2 percent) and a high incidence of neck recurrence (42 percent) were documented in those patients treated by primary tumor excision alone. Patients who underwent salvage neck dissection for recurrent neck node metastases had a 3 year survival rate of 56 percent. This study has documented a high incidence of cervical node recurrence in patients with T1 and T2 squamous cell carcinomas of the oral cavity treated by primary tumor excision alone and a poor survival rate after salvage therapy. A small group of patients who underwent elective neck dissection had a demonstrably high survival rate. These observations lend support to the call for elective neck dissection in patients with stage I and II oral cavity carcinoma but are not conclusive. Therapeutic decisions regarding elective treatment of the neck will continue to be made according to the best judgment and prejudices of the individual surgeon until a prospective, randomized multi-institutional study addressing this specific issue is undertaken. | 3,766,864 |
pubmed23n0125_13603 | The role of sternum in osteomyocutaneous reconstruction of major mandibular defects. | Replacement of skeletal support after extensive intraoral resection demands the use of a vascularized bone graft. This can be provided by microvascular free transfer using the ilium or metatarsal bones or by using muscle as a vascular carrier, employing the rib, the spine of the scapula, or the clavicle. Over a 5 year period, 27 patients underwent mandibular reconstruction with a vascularized segment of the outer table of sternum transposed together with a skin island on the pectoralis major muscle. Loss of bone occurred in two patients. A 93 percent survival rate was achieved. An earlier, smaller series of six patients using rib for reconstruction resulted in an unacceptably high complication rate. Success in survival of the bone graft, however, is not enough to procure a satisfactory result. Specific aspects of surgical technique are described that create an alveolar ridge with a sulcus on either side that can accept a denture; a diaphragm to replace the floor of the mouth and eliminate dead space; and an esthetically acceptable appearance, with definition of the jawline and a properly supported lower lip. | The role of sternum in osteomyocutaneous reconstruction of major mandibular defects. Replacement of skeletal support after extensive intraoral resection demands the use of a vascularized bone graft. This can be provided by microvascular free transfer using the ilium or metatarsal bones or by using muscle as a vascular carrier, employing the rib, the spine of the scapula, or the clavicle. Over a 5 year period, 27 patients underwent mandibular reconstruction with a vascularized segment of the outer table of sternum transposed together with a skin island on the pectoralis major muscle. Loss of bone occurred in two patients. A 93 percent survival rate was achieved. An earlier, smaller series of six patients using rib for reconstruction resulted in an unacceptably high complication rate. Success in survival of the bone graft, however, is not enough to procure a satisfactory result. Specific aspects of surgical technique are described that create an alveolar ridge with a sulcus on either side that can accept a denture; a diaphragm to replace the floor of the mouth and eliminate dead space; and an esthetically acceptable appearance, with definition of the jawline and a properly supported lower lip. | 3,766,865 |
pubmed23n0125_13604 | Prognostic factors in patients with regional cervical nodal metastases from cutaneous malignant melanoma. | A retrospective analysis with a minimum 10 year follow-up was performed on 287 patients who underwent radical or modified neck dissections with histologically involved regional nodal metastases from cutaneous malignant melanoma. The cumulative 5 year and 10 year survival rates calculated from the time of node dissection were 33 percent and 28 percent, respectively. Age and sex of the patient, site of known primary tumor, clinical stage at presentation, and time interval from the treatment of the primary tumor to node dissection did not independently affect survival. However, an unknown site of primary disease, the presence of only one histologically involved node, and the absence of extranodal tumor invasion at the time of node dissection were statistically significant individual prognostic factors for an improved survival rate. | Prognostic factors in patients with regional cervical nodal metastases from cutaneous malignant melanoma. A retrospective analysis with a minimum 10 year follow-up was performed on 287 patients who underwent radical or modified neck dissections with histologically involved regional nodal metastases from cutaneous malignant melanoma. The cumulative 5 year and 10 year survival rates calculated from the time of node dissection were 33 percent and 28 percent, respectively. Age and sex of the patient, site of known primary tumor, clinical stage at presentation, and time interval from the treatment of the primary tumor to node dissection did not independently affect survival. However, an unknown site of primary disease, the presence of only one histologically involved node, and the absence of extranodal tumor invasion at the time of node dissection were statistically significant individual prognostic factors for an improved survival rate. | 3,766,866 |
pubmed23n0125_13605 | Unique proteins defined by monoclonal antibodies specific for human melanoma. Some potential clinical applications. | A panel of monoclonal antibodies, designated HMB 18, 45, and 50, have been isolated that are highly specific for malignant melanoma. When tested on fixed, paraffin-embedded tissue sections, they reacted with 97 percent of melanomas tested (58 of 60), including pigmented, unpigmented, primary, and metastatic melanoma. The specificity in differentiating melanomas from other malignant tumors, including 112 carcinomas, 35 lymphomas, and 39 sarcomas, was 100 percent. Normal melanocytes were unreactive, although some benign melanocytic lesions were recognized. Using immunoprecipitation and SDS-PAGE analysis of 35S-methionine-labeled melanoma cells in tissue culture, a previously undescribed protein of approximately 10 kd was recognized by all three antibodies. HMB 50 also precipitated two high molecular weight proteins of 97 kd and 110 kd from the conditioned medium of melanoma cells. These monoclonal antibodies are the most sensitive and specific antibodies generated against human melanoma to date. Their clinical application in diagnostic surgical pathology and potential use in immunotherapy are discussed. | Unique proteins defined by monoclonal antibodies specific for human melanoma. Some potential clinical applications. A panel of monoclonal antibodies, designated HMB 18, 45, and 50, have been isolated that are highly specific for malignant melanoma. When tested on fixed, paraffin-embedded tissue sections, they reacted with 97 percent of melanomas tested (58 of 60), including pigmented, unpigmented, primary, and metastatic melanoma. The specificity in differentiating melanomas from other malignant tumors, including 112 carcinomas, 35 lymphomas, and 39 sarcomas, was 100 percent. Normal melanocytes were unreactive, although some benign melanocytic lesions were recognized. Using immunoprecipitation and SDS-PAGE analysis of 35S-methionine-labeled melanoma cells in tissue culture, a previously undescribed protein of approximately 10 kd was recognized by all three antibodies. HMB 50 also precipitated two high molecular weight proteins of 97 kd and 110 kd from the conditioned medium of melanoma cells. These monoclonal antibodies are the most sensitive and specific antibodies generated against human melanoma to date. Their clinical application in diagnostic surgical pathology and potential use in immunotherapy are discussed. | 3,766,867 |
pubmed23n0125_13606 | Soft tissue sarcomas of the head and neck in adolescents and adults. | Between 1960 and 1982, 188 patients were treated for soft tissue sarcomas of the head and neck. These patients had a heterogeneous group of neoplasms whose biologic behavior was determined by histologic classification, differentiation, and size. Histologic classification of these tumors was important and had prognostic significance. Differentiation affected local control and the propensity for distant metastases. Tumor size also contributed to outcome: patients with tumors of more than 5 cm had a worse survival than those with smaller sarcomas. Wide surgical excision with an adequate margin of normal tissue offered the best means of local control. The addition of postoperative radiotherapy was utilized for patients with positive margins or high grade aggressive sarcomas. Finally, despite multimodality therapy, achieving local control and prevention of distant disease in high grade sarcomas remains a major therapeutic challenge. | Soft tissue sarcomas of the head and neck in adolescents and adults. Between 1960 and 1982, 188 patients were treated for soft tissue sarcomas of the head and neck. These patients had a heterogeneous group of neoplasms whose biologic behavior was determined by histologic classification, differentiation, and size. Histologic classification of these tumors was important and had prognostic significance. Differentiation affected local control and the propensity for distant metastases. Tumor size also contributed to outcome: patients with tumors of more than 5 cm had a worse survival than those with smaller sarcomas. Wide surgical excision with an adequate margin of normal tissue offered the best means of local control. The addition of postoperative radiotherapy was utilized for patients with positive margins or high grade aggressive sarcomas. Finally, despite multimodality therapy, achieving local control and prevention of distant disease in high grade sarcomas remains a major therapeutic challenge. | 3,766,868 |
pubmed23n0125_13607 | Flow cytometric analysis of DNA content as a prognostic indicator in squamous cell carcinoma of the tongue. | The mortality of squamous cell carcinoma of the tongue has not significantly improved in decades. Much of the information that has been gathered to date has been based on retrospective analyses. There is little consensus on treatment of the disease. In an attempt to define an objective prognostic indicator of aggressiveness of these tumors, a retrospective analysis of 15 paraffin-embedded specimens using flow cytometry was performed. Ten patients (67 percent) had aneuploid tumors and had a 5 year disease-free survival rate of 33 percent, whereas patients with diploid tumors (33 percent) had a 5 year disease-free survival rate of 80 percent. Although the number of patients was small, it appears that flow cytometry may be an objective prognostic indicator in patients with squamous cell carcinoma of the tongue. Larger series of archival paraffin-embedded flow cytometry analyses are recommended, as well as examination of variables other than the disease-free survival rate. | Flow cytometric analysis of DNA content as a prognostic indicator in squamous cell carcinoma of the tongue. The mortality of squamous cell carcinoma of the tongue has not significantly improved in decades. Much of the information that has been gathered to date has been based on retrospective analyses. There is little consensus on treatment of the disease. In an attempt to define an objective prognostic indicator of aggressiveness of these tumors, a retrospective analysis of 15 paraffin-embedded specimens using flow cytometry was performed. Ten patients (67 percent) had aneuploid tumors and had a 5 year disease-free survival rate of 33 percent, whereas patients with diploid tumors (33 percent) had a 5 year disease-free survival rate of 80 percent. Although the number of patients was small, it appears that flow cytometry may be an objective prognostic indicator in patients with squamous cell carcinoma of the tongue. Larger series of archival paraffin-embedded flow cytometry analyses are recommended, as well as examination of variables other than the disease-free survival rate. | 3,766,869 |
pubmed23n0125_13608 | Significance of ploidy in laryngeal cancer. | We studied 48 patients with laryngeal or hypopharyngeal primary tumors with DNA flow cytometry. Twenty-four of the tumors were glottic, 18 were supraglottic, and 6 were from the pyriform sinus. Patients were followed for a minimum of 12 months, with a mean follow-up of 23 months. Twenty-three of the 48 primary tumors (48 percent) were clearly aneuploid, 22 percent were tetraploid, and 30 percent were diploid. We concluded that patients with aneuploid primary tumors, high DNA levels, or both have a significantly better prognosis than those with diploid tumors, and this remained statistically significant when clinical outcome was adjusted for tumor status, stage, and nodal status. | Significance of ploidy in laryngeal cancer. We studied 48 patients with laryngeal or hypopharyngeal primary tumors with DNA flow cytometry. Twenty-four of the tumors were glottic, 18 were supraglottic, and 6 were from the pyriform sinus. Patients were followed for a minimum of 12 months, with a mean follow-up of 23 months. Twenty-three of the 48 primary tumors (48 percent) were clearly aneuploid, 22 percent were tetraploid, and 30 percent were diploid. We concluded that patients with aneuploid primary tumors, high DNA levels, or both have a significantly better prognosis than those with diploid tumors, and this remained statistically significant when clinical outcome was adjusted for tumor status, stage, and nodal status. | 3,766,870 |
pubmed23n0125_13609 | Initial manifestation of acquired immunodeficiency syndrome in the head and neck region. | Initial manifestation of AIDS in the head and neck region occurs frequently. The purpose of this report has been to alert the head and neck surgeon to the occurrence of AIDS-related lesions, their clinical characteristics, and disease outcome. Incomplete recognition of these disorders may delay appropriate diagnostic study and initiation of therapy. We have described 10 patients in whom the initial manifestation of AIDS-related malignancies occurred in the head and neck region. Six of these patients were found to have Kaposi's sarcoma, whereas four had non-Hodgkin's lymphomas. The specific clinical and pathologic aspects of the disease have been described, which represent common patterns of presentation. It is crucial to obtain an accurate social history, as well as a complete medical history from any patient suspected of having AIDS, and prompt biopsy of suspect lesions should be performed. | Initial manifestation of acquired immunodeficiency syndrome in the head and neck region. Initial manifestation of AIDS in the head and neck region occurs frequently. The purpose of this report has been to alert the head and neck surgeon to the occurrence of AIDS-related lesions, their clinical characteristics, and disease outcome. Incomplete recognition of these disorders may delay appropriate diagnostic study and initiation of therapy. We have described 10 patients in whom the initial manifestation of AIDS-related malignancies occurred in the head and neck region. Six of these patients were found to have Kaposi's sarcoma, whereas four had non-Hodgkin's lymphomas. The specific clinical and pathologic aspects of the disease have been described, which represent common patterns of presentation. It is crucial to obtain an accurate social history, as well as a complete medical history from any patient suspected of having AIDS, and prompt biopsy of suspect lesions should be performed. | 3,766,871 |
pubmed23n0125_13610 | Extralaryngeal division of the recurrent laryngeal nerve. Report on 400 patients and the 721 nerves measured. | Seven hundred twenty-one recurrent laryngeal nerves were visualized in 400 patients having thyroid or parathyroid surgery. Four hundred twenty-one nerves (58 percent) bifurcated or trifurcated more than 0.5 cm from the cricoid cartilage. Ninety-seven patients had bilateral bifurcations, and 10 patients had trifurcations. Six patients had direct laryngeal nerves, all on the right side. One patient had a direct laryngeal and a recurrent laryngeal nerve simultaneously, and one patient had a bifurcated recurrent laryngeal nerve with an accessory vagus nerve joining it 13.5 cm from the cricoid cartilage. Damage to any of the filaments of the recurrent laryngeal nerve to the cricothyroid musculature or to or from a variant direct laryngeal nerve or variant vagus nerve connection can cause vocal cord paralysis. Damage to any branches of the recurrent laryngeal nerve to the esophagus can cause dysphagia. Therefore, if possible, all branches of the recurrent laryngeal nerve, no matter how small, should be preserved. | Extralaryngeal division of the recurrent laryngeal nerve. Report on 400 patients and the 721 nerves measured. Seven hundred twenty-one recurrent laryngeal nerves were visualized in 400 patients having thyroid or parathyroid surgery. Four hundred twenty-one nerves (58 percent) bifurcated or trifurcated more than 0.5 cm from the cricoid cartilage. Ninety-seven patients had bilateral bifurcations, and 10 patients had trifurcations. Six patients had direct laryngeal nerves, all on the right side. One patient had a direct laryngeal and a recurrent laryngeal nerve simultaneously, and one patient had a bifurcated recurrent laryngeal nerve with an accessory vagus nerve joining it 13.5 cm from the cricoid cartilage. Damage to any of the filaments of the recurrent laryngeal nerve to the cricothyroid musculature or to or from a variant direct laryngeal nerve or variant vagus nerve connection can cause vocal cord paralysis. Damage to any branches of the recurrent laryngeal nerve to the esophagus can cause dysphagia. Therefore, if possible, all branches of the recurrent laryngeal nerve, no matter how small, should be preserved. | 3,766,872 |
pubmed23n0125_13611 | Diagnostic accuracy of fine-needle aspiration biopsy versus frozen section in solitary thyroid nodules. | Fine-needle aspiration biopsy is widely used in the diagnosis and management of the solitary thyroid nodule. It is the most accurate tool available and decreases the need for ultrasonography and thyroid scanning. In those patients who are selected for surgical treatment by fine-needle aspiration biopsy, it has been advocated as a guide to determining the extent of operation. Frozen section, which usually serves as the surgeon's guide, and fine-needle aspiration biopsy both have varying accuracy rates. Few direct comparisons have been made. We studied 198 aspirates in 198 patients who presented with a solitary nodule and had surgical excision. Fine-needle aspiration biopsy (198 cases) and frozen section (182 cases) were compared with the final histologic diagnosis. Accuracy rates for fine-needle aspiration biopsy and frozen section were 90 percent and 95 percent, respectively. Fine-needle aspiration biopsy detected 43 percent of the cancers and frozen section, 64 percent. There were no false-positive diagnoses with frozen section, but three cases with fine-needle aspiration biopsy. The false-negative diagnosis rate was 5 percent for frozen section and 8.5 percent for fine-needle aspiration biopsy. When the "other" diagnosis category was grouped with the "positive" diagnosis category and a single expert cytopathologist was used to read the cytology report, the sensitivity was increased to 80 percent whereas the accuracy was maintained at 83 percent. We believe that since there were no false-positive diagnosis using frozen section, it can reliably be used as a guide when it reveals malignancy. When the fine-needle aspiration biopsy diagnosis is "positive" or "other," it can guide operation, but only after carefully assessing the wording of the cytology report and the clinical situation. Each modality can provide information missed by the other. We continue to use them as complementary tools in the diagnosis and management of solitary thyroid nodules. | Diagnostic accuracy of fine-needle aspiration biopsy versus frozen section in solitary thyroid nodules. Fine-needle aspiration biopsy is widely used in the diagnosis and management of the solitary thyroid nodule. It is the most accurate tool available and decreases the need for ultrasonography and thyroid scanning. In those patients who are selected for surgical treatment by fine-needle aspiration biopsy, it has been advocated as a guide to determining the extent of operation. Frozen section, which usually serves as the surgeon's guide, and fine-needle aspiration biopsy both have varying accuracy rates. Few direct comparisons have been made. We studied 198 aspirates in 198 patients who presented with a solitary nodule and had surgical excision. Fine-needle aspiration biopsy (198 cases) and frozen section (182 cases) were compared with the final histologic diagnosis. Accuracy rates for fine-needle aspiration biopsy and frozen section were 90 percent and 95 percent, respectively. Fine-needle aspiration biopsy detected 43 percent of the cancers and frozen section, 64 percent. There were no false-positive diagnoses with frozen section, but three cases with fine-needle aspiration biopsy. The false-negative diagnosis rate was 5 percent for frozen section and 8.5 percent for fine-needle aspiration biopsy. When the "other" diagnosis category was grouped with the "positive" diagnosis category and a single expert cytopathologist was used to read the cytology report, the sensitivity was increased to 80 percent whereas the accuracy was maintained at 83 percent. We believe that since there were no false-positive diagnosis using frozen section, it can reliably be used as a guide when it reveals malignancy. When the fine-needle aspiration biopsy diagnosis is "positive" or "other," it can guide operation, but only after carefully assessing the wording of the cytology report and the clinical situation. Each modality can provide information missed by the other. We continue to use them as complementary tools in the diagnosis and management of solitary thyroid nodules. | 3,766,873 |
pubmed23n0125_13612 | Outpatient thyroidectomy: a feasibility study. | In a group of 48 highly selected patients, thyroid surgery was performed on an outpatient basis. It was found to be a safe, cost-effective procedure, convenient for both the patient and the surgeon. Economic factors, although definitely a consideration, were not the compelling reason to perform thyroidectomy in this manner. | Outpatient thyroidectomy: a feasibility study. In a group of 48 highly selected patients, thyroid surgery was performed on an outpatient basis. It was found to be a safe, cost-effective procedure, convenient for both the patient and the surgeon. Economic factors, although definitely a consideration, were not the compelling reason to perform thyroidectomy in this manner. | 3,766,874 |
pubmed23n0125_13613 | Fine-needle aspiration in the diagnosis of cervical lymphadenopathy. | The extent of workup in patients with cervical adenopathy has always been controversial. Extensive workup in the absence of a histologic diagnosis indicative of a malignant process is unwarranted. Although open biopsy may be necessary for certain benign conditions, its routine application for metastatic nodes is not advised. We have used fine-needle aspiration as a routine procedure in the initial evaluation of cervical adenopathy. Over the past 3 years, 140 needle aspirations have been performed. Adequate specimens were obtained in 97 percent of the patients and the diagnostic accuracy was 96 percent. Overall, 45 percent of the patients had the diagnosis of a malignant tumor established whereas others had benign tumors. Metastatic squamous cell carcinoma was diagnosed accurately in all of the patients. Other malignant tumors reported accurately included lymphoma, adenocarcinoma, and metastatic thyroid carcinoma. Among the benign conditions, accurate diagnoses could be established in patients with tuberculosis, chronic lymphadenitis, and hyperplastic lymph nodes. The findings of fine-needle aspiration are helpful in directing subsequent workup. If the findings do not correlate with the clinical suspicion, greater weight is given to the clinical picture and diagnostic workup appropriate for the suspected disease is performed. Proper use of fine-needle aspiration requires close communication between an experienced cytologist and the head and neck surgeon. This series demonstrates that fine-needle aspiration is a safe, accurate, and valuable tool for the evaluation of cervical adenopathy. | Fine-needle aspiration in the diagnosis of cervical lymphadenopathy. The extent of workup in patients with cervical adenopathy has always been controversial. Extensive workup in the absence of a histologic diagnosis indicative of a malignant process is unwarranted. Although open biopsy may be necessary for certain benign conditions, its routine application for metastatic nodes is not advised. We have used fine-needle aspiration as a routine procedure in the initial evaluation of cervical adenopathy. Over the past 3 years, 140 needle aspirations have been performed. Adequate specimens were obtained in 97 percent of the patients and the diagnostic accuracy was 96 percent. Overall, 45 percent of the patients had the diagnosis of a malignant tumor established whereas others had benign tumors. Metastatic squamous cell carcinoma was diagnosed accurately in all of the patients. Other malignant tumors reported accurately included lymphoma, adenocarcinoma, and metastatic thyroid carcinoma. Among the benign conditions, accurate diagnoses could be established in patients with tuberculosis, chronic lymphadenitis, and hyperplastic lymph nodes. The findings of fine-needle aspiration are helpful in directing subsequent workup. If the findings do not correlate with the clinical suspicion, greater weight is given to the clinical picture and diagnostic workup appropriate for the suspected disease is performed. Proper use of fine-needle aspiration requires close communication between an experienced cytologist and the head and neck surgeon. This series demonstrates that fine-needle aspiration is a safe, accurate, and valuable tool for the evaluation of cervical adenopathy. | 3,766,875 |
pubmed23n0125_13614 | Surgical management of vascular malformations of the head and neck. | The evaluation and treatment of non-involuting hemangiomas with arteriovenous components and arteriovenous malformations of the head and neck remain difficult. Surgical excision is still the most important and effective method of controlling these lesions. Radiotherapy should never be used. It is ineffective, dangerous, and often complicates any later operation that may be required. Evaluation of vascular malformations with computerized tomography, arteriography, magnetic nuclear imaging, and Doppler mapping will aid diagnosis. Adjunctive hypotensive anesthesia, intraoperative embolization with Gelfoam, and temporary peripheral suture ligations have made the surgical task more manageable. The integration of reconstructive techniques into the surgical program is essential if optimum results are to be obtained. Incisions must be planned so that future flap patterns are preserved. Axial vessels may be needed later. Fascial slings, muscle transfers, nerve grafts, and tissue expansion of adjacent normal tissue may be needed to maintain and restore function and features. Above all, the treatment of each patient requires individual planning. The most common errors in the treatment of vascular hemangiomas result from missed diagnoses and faint-heartedness in tackling the surgical removal of such highly vascular tumors. Many patients go for years without finding a surgeon who will help them. We believe that aggressive surgical treatment offers much to many of these patients. | Surgical management of vascular malformations of the head and neck. The evaluation and treatment of non-involuting hemangiomas with arteriovenous components and arteriovenous malformations of the head and neck remain difficult. Surgical excision is still the most important and effective method of controlling these lesions. Radiotherapy should never be used. It is ineffective, dangerous, and often complicates any later operation that may be required. Evaluation of vascular malformations with computerized tomography, arteriography, magnetic nuclear imaging, and Doppler mapping will aid diagnosis. Adjunctive hypotensive anesthesia, intraoperative embolization with Gelfoam, and temporary peripheral suture ligations have made the surgical task more manageable. The integration of reconstructive techniques into the surgical program is essential if optimum results are to be obtained. Incisions must be planned so that future flap patterns are preserved. Axial vessels may be needed later. Fascial slings, muscle transfers, nerve grafts, and tissue expansion of adjacent normal tissue may be needed to maintain and restore function and features. Above all, the treatment of each patient requires individual planning. The most common errors in the treatment of vascular hemangiomas result from missed diagnoses and faint-heartedness in tackling the surgical removal of such highly vascular tumors. Many patients go for years without finding a surgeon who will help them. We believe that aggressive surgical treatment offers much to many of these patients. | 3,766,876 |
pubmed23n0125_13615 | Major head and neck reconstruction using the deltopectoral flap. A 20 year experience. | Six hundred seventy-eight deltopectoral flaps were raised in 604 patients, 125 of which were delayed and 215 of which were used in previously irradiated beds. The rate of major flap necrosis was 16.9 percent and the overall rate of complications, 51.4 percent. Delay in creating the deltopectoral flap had no influence on the risk of complications and necrosis, whereas the use of the flap in a previously irradiated bed was associated with a significantly increased risk of major flap necrosis. The least flap loss occurred when the deltopectoral flap was used without tubulation for skin coverage only. Complications and flap necrosis occurred most frequently when flaps were tubulated in a reversed manner or used for lining of major portions of or for total oropharyngeal and hypopharyngeal reconstruction. The deltopectoral flap remains a useful, reliable, and versatile regional flap that can be used alone or in combination with other flaps in selected circumstances for major head and neck reconstruction. | Major head and neck reconstruction using the deltopectoral flap. A 20 year experience. Six hundred seventy-eight deltopectoral flaps were raised in 604 patients, 125 of which were delayed and 215 of which were used in previously irradiated beds. The rate of major flap necrosis was 16.9 percent and the overall rate of complications, 51.4 percent. Delay in creating the deltopectoral flap had no influence on the risk of complications and necrosis, whereas the use of the flap in a previously irradiated bed was associated with a significantly increased risk of major flap necrosis. The least flap loss occurred when the deltopectoral flap was used without tubulation for skin coverage only. Complications and flap necrosis occurred most frequently when flaps were tubulated in a reversed manner or used for lining of major portions of or for total oropharyngeal and hypopharyngeal reconstruction. The deltopectoral flap remains a useful, reliable, and versatile regional flap that can be used alone or in combination with other flaps in selected circumstances for major head and neck reconstruction. | 3,766,877 |
pubmed23n0125_13616 | Cheek island flap for replacement of critical limited defects of the upper aerodigestive tract. | Our experience thus far indicates that the island cheek flap is a satisfactory reconstructive method for small but critical defects in the upper aerodigestive tract. The mucosal lining is an ideal replacement for the lining of the mouth, pharynx, and esophagus. Only a single stage procedure is required. The surgical technique is relatively simple and does not unnecessarily prolong an extensive operation for tumor resection. We have found the procedure to be reliable with no instances of flap loss or necrosis. | Cheek island flap for replacement of critical limited defects of the upper aerodigestive tract. Our experience thus far indicates that the island cheek flap is a satisfactory reconstructive method for small but critical defects in the upper aerodigestive tract. The mucosal lining is an ideal replacement for the lining of the mouth, pharynx, and esophagus. Only a single stage procedure is required. The surgical technique is relatively simple and does not unnecessarily prolong an extensive operation for tumor resection. We have found the procedure to be reliable with no instances of flap loss or necrosis. | 3,766,878 |
pubmed23n0125_13617 | Craniofacial reconstruction after tumor resections using vascularized outer table calvarial bone flaps. | The outer table of calvarium is a useful donor site for facial reconstruction after resection of tumors. Large defects, especially when associated with multiple operations and radiotherapy, are poor recipient beds for nonvascularized bone grafts. A technique for the transfer of vascularized outer table has been developed and used in 11 patients for reconstruction of tumor defects. Satisfactory correction of both functional and esthetic problems has been achieved with satisfactory long-term stability and low morbidity. | Craniofacial reconstruction after tumor resections using vascularized outer table calvarial bone flaps. The outer table of calvarium is a useful donor site for facial reconstruction after resection of tumors. Large defects, especially when associated with multiple operations and radiotherapy, are poor recipient beds for nonvascularized bone grafts. A technique for the transfer of vascularized outer table has been developed and used in 11 patients for reconstruction of tumor defects. Satisfactory correction of both functional and esthetic problems has been achieved with satisfactory long-term stability and low morbidity. | 3,766,879 |
pubmed23n0125_13618 | Preoperative irradiation potentiation with cisplatin: effect on rate of wound infection. | Platinum coordination complexes, such as cisplatin, potentiate the cytotoxicity of irradiation on squamous cell carcinoma and certain other solid tumors. Using a rat oro-cutaneous fistula model, an investigation was carried out to determine whether or not there was a concomitant potentiation with cisplatin of the deleterious effect of preoperative irradiation on the ability of a subsequent wound to handle a bacterial challenge. Auto-contaminated wounds were found to have increased rates of infection at single-dose orthovoltage pretreatments of 1,500 rads or more. Using quantitative bacteriologic techniques, would infection was found to be no more frequent after platinum-enhanced irradiation than after irradiation alone; however, there was the additive effect of weight loss associated with combined cisplatin treatment and irradiation. | Preoperative irradiation potentiation with cisplatin: effect on rate of wound infection. Platinum coordination complexes, such as cisplatin, potentiate the cytotoxicity of irradiation on squamous cell carcinoma and certain other solid tumors. Using a rat oro-cutaneous fistula model, an investigation was carried out to determine whether or not there was a concomitant potentiation with cisplatin of the deleterious effect of preoperative irradiation on the ability of a subsequent wound to handle a bacterial challenge. Auto-contaminated wounds were found to have increased rates of infection at single-dose orthovoltage pretreatments of 1,500 rads or more. Using quantitative bacteriologic techniques, would infection was found to be no more frequent after platinum-enhanced irradiation than after irradiation alone; however, there was the additive effect of weight loss associated with combined cisplatin treatment and irradiation. | 3,766,880 |
pubmed23n0125_13619 | Neck dissection with and without radiotherapy: prognostic factors, patterns of recurrence, and survival. | A group of 179 patients who had 205 neck dissections between 1979 and 1984 has been reviewed to assess the influence of adjuvant radiotherapy on survival. Lymph nodes were histologically involved in 91 of 107 radical neck dissections (85 percent) and 55 of 98 modified neck dissections (56 percent). Eighty-two patients received adjuvant radiotherapy of 5,000 rads or more. Patients with involved nodes had significantly lower survival rates than those with uninvolved nodes. Among patients with involved nodes, survival was significantly lower when two or more nodes were involved, when there was nodal involvement at multiple levels, or when extracapsular spread was present. Adjuvant radiotherapy was associated with a reduced recurrence rate in the ipsilateral neck but the incidence of distant metastases was higher. When patients with involved nodes were subgrouped according to prognostic factors, the survival of irradiated patients was improved only in the highest risk group, but this was not statistically significant. When radiotherapy is added to neck dissection for treatment of cervical metastases it can be expected to reduced ipsilateral neck recurrence and prevent relapse in the contralateral neck. Improved survival may depend on an ability to detect and treat occult distant metastases. | Neck dissection with and without radiotherapy: prognostic factors, patterns of recurrence, and survival. A group of 179 patients who had 205 neck dissections between 1979 and 1984 has been reviewed to assess the influence of adjuvant radiotherapy on survival. Lymph nodes were histologically involved in 91 of 107 radical neck dissections (85 percent) and 55 of 98 modified neck dissections (56 percent). Eighty-two patients received adjuvant radiotherapy of 5,000 rads or more. Patients with involved nodes had significantly lower survival rates than those with uninvolved nodes. Among patients with involved nodes, survival was significantly lower when two or more nodes were involved, when there was nodal involvement at multiple levels, or when extracapsular spread was present. Adjuvant radiotherapy was associated with a reduced recurrence rate in the ipsilateral neck but the incidence of distant metastases was higher. When patients with involved nodes were subgrouped according to prognostic factors, the survival of irradiated patients was improved only in the highest risk group, but this was not statistically significant. When radiotherapy is added to neck dissection for treatment of cervical metastases it can be expected to reduced ipsilateral neck recurrence and prevent relapse in the contralateral neck. Improved survival may depend on an ability to detect and treat occult distant metastases. | 3,766,881 |
pubmed23n0125_13620 | Primary tracheoesophageal puncture for voice restoration. | A 93 percent success rate of voice restoration after primary tracheoesophageal puncture was achieved in 15 patients. Patients were usually able to talk within 3 weeks after laryngectomy. Male patients without chronic obstructive lung disease achieved better results. The incidence of complication after this procedure were not significantly different from a group of 111 patients who underwent laryngectomy alone. Finally, early speech acquisition provides a substantial psychologic boost for the patient undergoing total laryngectomy. | Primary tracheoesophageal puncture for voice restoration. A 93 percent success rate of voice restoration after primary tracheoesophageal puncture was achieved in 15 patients. Patients were usually able to talk within 3 weeks after laryngectomy. Male patients without chronic obstructive lung disease achieved better results. The incidence of complication after this procedure were not significantly different from a group of 111 patients who underwent laryngectomy alone. Finally, early speech acquisition provides a substantial psychologic boost for the patient undergoing total laryngectomy. | 3,766,882 |
pubmed23n0125_13621 | Pharyngeal carcinoma and natural killer cell activity. | An evaluation of natural killer cell activity was performed in 42 patients with pharyngeal carcinoma. Compared with age- and sex-matched control subjects, the cancer patients expressed significantly lower cytotoxicity against K562 target cells (68 +/- 8 lytic units versus 99 +/- 8 lytic units, p less than 0.01), with 52 percent of the patients expressing deficient activity (below 1 standard deviation of the mean activity of the control population). The probability of deficient activity was greater in these patients than observed in patients with cancer of other head and neck sites. Although natural killer cell activity was lower in patients who drank alcohol or had nodal metastases, no single clinical factor was predictive of deficient cytotoxic response. Prospective longitudinal evaluation (mean = 12 months) of these pharyngeal cancer patients demonstrated that deficient natural killer cell activity measured before treatment identified a population with a significantly increased risk for the development of distant metastases. Distant metastases developed in 7 of 18 patients (39 percent) with deficient natural killer cell activity. In contrast, none of the 16 patients with normal natural killer cell function had evidence of distant disease at last follow-up (p less than 0.01). Deficient natural killer cell activity exists in patients with pharyngeal cancer and is an independent marker for the subsequent development of distant metastases. | Pharyngeal carcinoma and natural killer cell activity. An evaluation of natural killer cell activity was performed in 42 patients with pharyngeal carcinoma. Compared with age- and sex-matched control subjects, the cancer patients expressed significantly lower cytotoxicity against K562 target cells (68 +/- 8 lytic units versus 99 +/- 8 lytic units, p less than 0.01), with 52 percent of the patients expressing deficient activity (below 1 standard deviation of the mean activity of the control population). The probability of deficient activity was greater in these patients than observed in patients with cancer of other head and neck sites. Although natural killer cell activity was lower in patients who drank alcohol or had nodal metastases, no single clinical factor was predictive of deficient cytotoxic response. Prospective longitudinal evaluation (mean = 12 months) of these pharyngeal cancer patients demonstrated that deficient natural killer cell activity measured before treatment identified a population with a significantly increased risk for the development of distant metastases. Distant metastases developed in 7 of 18 patients (39 percent) with deficient natural killer cell activity. In contrast, none of the 16 patients with normal natural killer cell function had evidence of distant disease at last follow-up (p less than 0.01). Deficient natural killer cell activity exists in patients with pharyngeal cancer and is an independent marker for the subsequent development of distant metastases. | 3,766,883 |
pubmed23n0125_13622 | Syndrome of septo-optic-pituitary dysplasia: the clinical spectrum. | Septo-optic-pituitary dysplasia is a syndrome characterized by abnormalities of midline brain structures, optic nerve hypoplasia, and congenital hypothalamic-pituitary insufficiency. Four infants, diagnosed as having clinical variations of this disorder, are described. The first had agenesis of the septum pellucidum and corpus callosum, partial hypothalamic insufficiency, and normal optic nerves. The second had a structurally normal brain, bilateral optic nerve hypoplasia, and panhypopituitarism. The third had bilateral agenesis of the cerebral cortex, bilateral optic nerve hypoplasia and partial hypothalamic insufficiency. The fourth had a structurally normal brain, unilateral optic nerve hypoplasia and panhypopituitarism. A review of the recent literature revealed 191 patients with bilateral optic nerve hypoplasia who were examined for possible existence of this syndrome. Of the 178 patients who had radiographic imaging of the brain, 60% were found to have structural abnormalities. Of the 145 patients evaluated for pituitary function, 62% had evidence of insufficiency. Approximately 30% of fully evaluated patients had evidence of all three components of the syndrome. Because of high incidence of structural abnormalities of the brain and congenital hypopituitarism in patients with optic nerve hypoplasia, we conclude that full evaluation is indicated in patients with any of the three components of the syndrome. | Syndrome of septo-optic-pituitary dysplasia: the clinical spectrum. Septo-optic-pituitary dysplasia is a syndrome characterized by abnormalities of midline brain structures, optic nerve hypoplasia, and congenital hypothalamic-pituitary insufficiency. Four infants, diagnosed as having clinical variations of this disorder, are described. The first had agenesis of the septum pellucidum and corpus callosum, partial hypothalamic insufficiency, and normal optic nerves. The second had a structurally normal brain, bilateral optic nerve hypoplasia, and panhypopituitarism. The third had bilateral agenesis of the cerebral cortex, bilateral optic nerve hypoplasia and partial hypothalamic insufficiency. The fourth had a structurally normal brain, unilateral optic nerve hypoplasia and panhypopituitarism. A review of the recent literature revealed 191 patients with bilateral optic nerve hypoplasia who were examined for possible existence of this syndrome. Of the 178 patients who had radiographic imaging of the brain, 60% were found to have structural abnormalities. Of the 145 patients evaluated for pituitary function, 62% had evidence of insufficiency. Approximately 30% of fully evaluated patients had evidence of all three components of the syndrome. Because of high incidence of structural abnormalities of the brain and congenital hypopituitarism in patients with optic nerve hypoplasia, we conclude that full evaluation is indicated in patients with any of the three components of the syndrome. | 3,766,900 |
pubmed23n0125_13623 | The effect of neonatal hyperbilirubinemia on the auditory brainstem response. | Auditory brainstem response (ABR) testing was performed on 30 babies in the Premature Baby Center of Kyoto University Hospital as soon after birth as possible. In cases of hyperbilirubinemia of more than 15.0 mg/dl, ABR testing was performed at least twice. In four babies with total bilirubin concentrations of more than 18.0 mg/dl at some point, abnormal ABRs were seen. These abnormalities normalized with a decrease in serum total bilirubin concentration in two of the four cases, but in the other two the abnormalities showed no change. The cause and significance of this difference are not known, but ABR is a simple and useful method for assessing the neurotoxic effects of bilirubin on the nervous system. | The effect of neonatal hyperbilirubinemia on the auditory brainstem response. Auditory brainstem response (ABR) testing was performed on 30 babies in the Premature Baby Center of Kyoto University Hospital as soon after birth as possible. In cases of hyperbilirubinemia of more than 15.0 mg/dl, ABR testing was performed at least twice. In four babies with total bilirubin concentrations of more than 18.0 mg/dl at some point, abnormal ABRs were seen. These abnormalities normalized with a decrease in serum total bilirubin concentration in two of the four cases, but in the other two the abnormalities showed no change. The cause and significance of this difference are not known, but ABR is a simple and useful method for assessing the neurotoxic effects of bilirubin on the nervous system. | 3,766,901 |
pubmed23n0125_13624 | Serum and urinary carnitine and organic acids in Reye syndrome and Reye-like syndrome. | Free and acyl-carnitine in serum and urine, and urinary organic acids were measured in 6 patients with Reye syndrome and Reye-like syndrome. The free and total carnitine concentrations were significantly reduced in serum during the acute phases of the diseases. Thus, the ratio of acylcarnitine to free carnitine was significantly increased. Urinary excretion of acylcarnitine was greatly increased, and the acylcarnitine to total carnitine ratio was therefore greater than in controls. The urinary organic acids comprised large amounts of lactic acid, dicarboxylic acids and ketone bodies. It is suggested that carnitine deficiency is induced as more carnitine is consumed to buffer the increased amount of toxic acyl-CoA compounds metabolized from free fatty acids and the many organic acids. These results indicate that administration of L-carnitine should generally be considered in patients with Reye syndrome and Reye-like syndrome. | Serum and urinary carnitine and organic acids in Reye syndrome and Reye-like syndrome. Free and acyl-carnitine in serum and urine, and urinary organic acids were measured in 6 patients with Reye syndrome and Reye-like syndrome. The free and total carnitine concentrations were significantly reduced in serum during the acute phases of the diseases. Thus, the ratio of acylcarnitine to free carnitine was significantly increased. Urinary excretion of acylcarnitine was greatly increased, and the acylcarnitine to total carnitine ratio was therefore greater than in controls. The urinary organic acids comprised large amounts of lactic acid, dicarboxylic acids and ketone bodies. It is suggested that carnitine deficiency is induced as more carnitine is consumed to buffer the increased amount of toxic acyl-CoA compounds metabolized from free fatty acids and the many organic acids. These results indicate that administration of L-carnitine should generally be considered in patients with Reye syndrome and Reye-like syndrome. | 3,766,902 |
pubmed23n0125_13625 | Hypoxic-ischemic brain damage and cerebral blood flow changes in young rabbits. | Hypoxic-ischemic cerebral damage was demonstrated in the cerebral cortex of 2-week-old rabbits at 3 to 5 days after bilateral carotid artery ligation and reperfusion during hypoxemia. Carotid ligation and reperfusion had little effect on cortical blood flow during normoxemia, but the former suppressed a blood flow increase to hypoxemia and the latter suppressed a blood flow decrease to hyperoxemia. These results suggest a relative ischemia or vascular dysfunction which may play a part in the pathogenesis of the hypoxic-ischemic cortical necrosis. | Hypoxic-ischemic brain damage and cerebral blood flow changes in young rabbits. Hypoxic-ischemic cerebral damage was demonstrated in the cerebral cortex of 2-week-old rabbits at 3 to 5 days after bilateral carotid artery ligation and reperfusion during hypoxemia. Carotid ligation and reperfusion had little effect on cortical blood flow during normoxemia, but the former suppressed a blood flow increase to hypoxemia and the latter suppressed a blood flow decrease to hyperoxemia. These results suggest a relative ischemia or vascular dysfunction which may play a part in the pathogenesis of the hypoxic-ischemic cortical necrosis. | 3,766,903 |
pubmed23n0125_13626 | Ten years follow up study of steroid therapy for congenital encephalomyopathy. | Two 10-year-old boys with mental retardation and myopathy which were present since birth are described. Both had elevated serum creatine phosphokinase (CK) and one of them had a positive family history. The clinical features were consistent with Fukuyama type congenital muscular dystrophy, but muscle biopsies suggested an inflammatory process. Adrenal cortical steroids were given and they were followed up until 10 years of age. Serum CK showed a significant response to the treatment, and mental retardation in case 1 and motor dysfunction in case 2 improved. It is postulated that an inflammatory process might be a causative factor in some patients with congenital muscular dystrophy. | Ten years follow up study of steroid therapy for congenital encephalomyopathy. Two 10-year-old boys with mental retardation and myopathy which were present since birth are described. Both had elevated serum creatine phosphokinase (CK) and one of them had a positive family history. The clinical features were consistent with Fukuyama type congenital muscular dystrophy, but muscle biopsies suggested an inflammatory process. Adrenal cortical steroids were given and they were followed up until 10 years of age. Serum CK showed a significant response to the treatment, and mental retardation in case 1 and motor dysfunction in case 2 improved. It is postulated that an inflammatory process might be a causative factor in some patients with congenital muscular dystrophy. | 3,766,904 |
pubmed23n0125_13627 | Progressive muscular dystrophy with congenital adrenal hypoplasia: an unusual autopsy case. | A 3 1/2-year-old child with progressive muscular dystrophy (PMD) and congenital adrenal hypoplasia (CAH) is described. Symptoms and signs of adrenocortical insufficiency appeared shortly after birth. Despite corticosteroid therapy, the muscular weakness and elevated CK level continued. A diagnosis of Duchenne muscular dystrophy was made on the basis of clinical signs and characteristic muscle biopsy. The affection of his older brother suggests an X-linked recessive inheritance. The autopsy revealed a very rare combination of cytomegalic type CAH and PMD. This combination suggests that a small deletion of X-chromosome might be responsible for the two disorders. | Progressive muscular dystrophy with congenital adrenal hypoplasia: an unusual autopsy case. A 3 1/2-year-old child with progressive muscular dystrophy (PMD) and congenital adrenal hypoplasia (CAH) is described. Symptoms and signs of adrenocortical insufficiency appeared shortly after birth. Despite corticosteroid therapy, the muscular weakness and elevated CK level continued. A diagnosis of Duchenne muscular dystrophy was made on the basis of clinical signs and characteristic muscle biopsy. The affection of his older brother suggests an X-linked recessive inheritance. The autopsy revealed a very rare combination of cytomegalic type CAH and PMD. This combination suggests that a small deletion of X-chromosome might be responsible for the two disorders. | 3,766,905 |
pubmed23n0125_13628 | The protective effects of vitamin E against microcephaly in rats X-irradiated in utero: dendritic branches. | Fetuses from rats given either water or 0.03% dl-alpha-tocopherol acetate (vitamin E) as the drinking fluid and X-irradiated with 100R on gestational day (gd) 13 were examined on gd 21. The degree of dendritic branching of frontal cerebral neurons, examined after Golgi staining, was significantly reduced by X-irradiation, and supplementation of vitamin E caused increased branching order numbers in irradiated fetuses. This study provides evidence of the protection by vitamin E of neuronal development in X-irradiated fetuses. | The protective effects of vitamin E against microcephaly in rats X-irradiated in utero: dendritic branches. Fetuses from rats given either water or 0.03% dl-alpha-tocopherol acetate (vitamin E) as the drinking fluid and X-irradiated with 100R on gestational day (gd) 13 were examined on gd 21. The degree of dendritic branching of frontal cerebral neurons, examined after Golgi staining, was significantly reduced by X-irradiation, and supplementation of vitamin E caused increased branching order numbers in irradiated fetuses. This study provides evidence of the protection by vitamin E of neuronal development in X-irradiated fetuses. | 3,766,906 |
pubmed23n0125_13629 | Fanconi's anemia. Clinical study of six cases. | Six cases of constitutional aplastic anemia (Fanconi's anemia), including four boys and two girls who were between 6 and 10 years of age, are presented. Congenital abnormalities were observed in all the patients, especially skin pigmentations, renal anomalies, and growth retardation. The initial symptoms were insidious, mainly related to the anemia. All patients were anemic and thrombocytopenic, but only three were leukopenic. Five cases showed increased levels of fetal hemoglobin, which was marked in two. Bone marrow obtained by aspiration and/or biopsy was hypocellular in all cases. Chromosomal studies showed breaks, ring- or cross-chromosomes, chromosome exchanges, and endoreduplications. Five cases were treated with androgens and a good response was obtained in all of them. Four patients died, and the remaining two are alive and well. The clinical and biological features of these patients are in accordance with those of previous reports. The importance of the androgen therapy is stressed; androgen dependence was observed in all cases. | Fanconi's anemia. Clinical study of six cases. Six cases of constitutional aplastic anemia (Fanconi's anemia), including four boys and two girls who were between 6 and 10 years of age, are presented. Congenital abnormalities were observed in all the patients, especially skin pigmentations, renal anomalies, and growth retardation. The initial symptoms were insidious, mainly related to the anemia. All patients were anemic and thrombocytopenic, but only three were leukopenic. Five cases showed increased levels of fetal hemoglobin, which was marked in two. Bone marrow obtained by aspiration and/or biopsy was hypocellular in all cases. Chromosomal studies showed breaks, ring- or cross-chromosomes, chromosome exchanges, and endoreduplications. Five cases were treated with androgens and a good response was obtained in all of them. Four patients died, and the remaining two are alive and well. The clinical and biological features of these patients are in accordance with those of previous reports. The importance of the androgen therapy is stressed; androgen dependence was observed in all cases. | 3,766,907 |
pubmed23n0125_13630 | Clinical and in vitro antiproliferative properties of recombinant DNA-derived human interferon-alpha 2. | The properties of the recombinant DNA-derived human leukocyte interferon, HuIFN alpha 2, were studied in patients with advanced leukemia or lymphoma. In vitro, HuIFN alpha 2 induced an increased activity of 2-5A synthetase in leukemic and in control cells indicating cellular responsiveness to IFN. HuIFN alpha 2 also produced a dose-responsive decline in marrow leukemia blast progenitor colony growth, and in normal hematopoietic colony formation in vitro, confirming its antiproliferative effect. A course of intravenous therapy given to a lymphoma patient produced a modest decline in peripheral white blood cell (WBC) and neutrophil counts; higher, more frequent doses in a second patient induced a profound drop in WBC's, neutrophils, and platelets. When the leukemia patients were given an intravenous course of HuIFN alpha 2 as a sole agent, blast cytoreduction was seen in peripheral blood in three patients, and in marrow of one patient with acute myeloblastic leukemia (AML). Elevated 2-5A synthetase levels could be detected after therapy. No modulation of leukemic cell markers was seen after in vitro or in vivo treatment with HuIFN alpha 2, implying that the cytoreduction was not linked to blast cell differentiation. These studies suggest that this subtype of recombinant DNA-derived IFN has antileukemic properties, and indicates the possibilities for IFN as an adjunctive form of therapy in leukemia. | Clinical and in vitro antiproliferative properties of recombinant DNA-derived human interferon-alpha 2. The properties of the recombinant DNA-derived human leukocyte interferon, HuIFN alpha 2, were studied in patients with advanced leukemia or lymphoma. In vitro, HuIFN alpha 2 induced an increased activity of 2-5A synthetase in leukemic and in control cells indicating cellular responsiveness to IFN. HuIFN alpha 2 also produced a dose-responsive decline in marrow leukemia blast progenitor colony growth, and in normal hematopoietic colony formation in vitro, confirming its antiproliferative effect. A course of intravenous therapy given to a lymphoma patient produced a modest decline in peripheral white blood cell (WBC) and neutrophil counts; higher, more frequent doses in a second patient induced a profound drop in WBC's, neutrophils, and platelets. When the leukemia patients were given an intravenous course of HuIFN alpha 2 as a sole agent, blast cytoreduction was seen in peripheral blood in three patients, and in marrow of one patient with acute myeloblastic leukemia (AML). Elevated 2-5A synthetase levels could be detected after therapy. No modulation of leukemic cell markers was seen after in vitro or in vivo treatment with HuIFN alpha 2, implying that the cytoreduction was not linked to blast cell differentiation. These studies suggest that this subtype of recombinant DNA-derived IFN has antileukemic properties, and indicates the possibilities for IFN as an adjunctive form of therapy in leukemia. | 3,766,908 |
pubmed23n0125_13631 | Burkitt's lymphoma. Experience at the Instituto Nacional de Pediatria, Mexico City. | Thirty patients with Burkitt's lymphoma sporadic type are presented. All of these cases fulfilled the histologic criteria of Burkitt's tumor. The age ranged from 2 to 12 years, with a median of 4 years. Fever and chills were the most common symptoms. The abdomen was the site of initial presentation in more than half of these cases. Close to one-third of the patients had involvement of the central nervous system. Fifty percent of the patients presented with lactic dehydrogenase elevation. Uric acid was elevated in close to one-fourth of these patients. The sonogram was the most useful technique in those instances of abdominal presentation. All patients were staged according to Ziegler criteria after an initial surgical diagnostic procedure was performed. None of the patients received radiotherapy to the primary tumor. All were submitted to a chemotherapy program for 12 months. Only 26 fitted the criteria of being evaluable. The duration of remission was from 1 to 144 months, with a median of 18.2 months. The actuarial survival was 84% at 3 years. Most of the responders corresponded to Stages A, B, and AR, while Stages C and D had a high mortality rate with elevated lactic dehydrogenase and uric acid levels. | Burkitt's lymphoma. Experience at the Instituto Nacional de Pediatria, Mexico City. Thirty patients with Burkitt's lymphoma sporadic type are presented. All of these cases fulfilled the histologic criteria of Burkitt's tumor. The age ranged from 2 to 12 years, with a median of 4 years. Fever and chills were the most common symptoms. The abdomen was the site of initial presentation in more than half of these cases. Close to one-third of the patients had involvement of the central nervous system. Fifty percent of the patients presented with lactic dehydrogenase elevation. Uric acid was elevated in close to one-fourth of these patients. The sonogram was the most useful technique in those instances of abdominal presentation. All patients were staged according to Ziegler criteria after an initial surgical diagnostic procedure was performed. None of the patients received radiotherapy to the primary tumor. All were submitted to a chemotherapy program for 12 months. Only 26 fitted the criteria of being evaluable. The duration of remission was from 1 to 144 months, with a median of 18.2 months. The actuarial survival was 84% at 3 years. Most of the responders corresponded to Stages A, B, and AR, while Stages C and D had a high mortality rate with elevated lactic dehydrogenase and uric acid levels. | 3,766,909 |
pubmed23n0125_13632 | Acute leukemia in infants. A unique pattern of acute nonlymphocytic leukemia. | At present, the majority of children with acute lymphoblastic leukemia can expect a favorable outcome. However, leukemia in infants (0-1 year) still runs a dismal course. The first year of life also has a unique pattern of hematopoietic activity. We investigated possible association between these two findings. We examined the characteristics of 51 infants with leukemia, diagnosed from 1975 to 1984; 48/51 had evaluable data. Infants had a high frequency (39.6%) of hyperleukocytosis (greater than 100 X 10(9)/liter), early central nervous system involvement (27.1%), and cutaneous infiltration (18.8%). The respective numbers in patients greater than or equal to 1 year were 13.9, 4.8, and 1.0%. Acute nonlymphocytic leukemia was diagnosed in 18/48 infants (37.5%) compared to 128/892 (14.3%) in older patients. Infants had only acute leukemia of French-American-British (FAB) class M5 (10/19) or acute leukemia with unclassifiable but monocytoid blasts (8/19). This study confirms the presence of poor prognostic features in infant leukemia. The almost exclusive occurrence of monoblastic or monocytoid acute leukemia parallels the high proliferation rate of monocytes in this age group and the leukemic cells may thus well reflect frozen stages of monocytic differentiation. | Acute leukemia in infants. A unique pattern of acute nonlymphocytic leukemia. At present, the majority of children with acute lymphoblastic leukemia can expect a favorable outcome. However, leukemia in infants (0-1 year) still runs a dismal course. The first year of life also has a unique pattern of hematopoietic activity. We investigated possible association between these two findings. We examined the characteristics of 51 infants with leukemia, diagnosed from 1975 to 1984; 48/51 had evaluable data. Infants had a high frequency (39.6%) of hyperleukocytosis (greater than 100 X 10(9)/liter), early central nervous system involvement (27.1%), and cutaneous infiltration (18.8%). The respective numbers in patients greater than or equal to 1 year were 13.9, 4.8, and 1.0%. Acute nonlymphocytic leukemia was diagnosed in 18/48 infants (37.5%) compared to 128/892 (14.3%) in older patients. Infants had only acute leukemia of French-American-British (FAB) class M5 (10/19) or acute leukemia with unclassifiable but monocytoid blasts (8/19). This study confirms the presence of poor prognostic features in infant leukemia. The almost exclusive occurrence of monoblastic or monocytoid acute leukemia parallels the high proliferation rate of monocytes in this age group and the leukemic cells may thus well reflect frozen stages of monocytic differentiation. | 3,766,910 |
pubmed23n0125_13633 | Aggressive combination therapy in the successful management of life-threatening intracranial hemorrhage in a patient with idiopathic thrombocytopenic purpura. | We describe acute therapy for a 13-year-old female from Panama with chronic idiopathic thrombocytopenia purpura (ITP) refractory to steroids, splenectomy, vinca alkaloids, and azathioprine. She presented with neurologic deterioration from a posterior fossa intracranial hemorrhage (ICH). This followed a 3-month history of severe dysfunctional uterine bleeding, progressive from menarche, which had required multiple red cell transfusions. Steroid and vinblastine therapy and transfusion of 40 U of platelets failed to increase the platelet count above 10,000/microliter. Development of a second larger ICH (frontal) produced morbid increase in intracranial pressure that necessitated neurosurgical decompression. Plasma exchange and colloid repletion with intravenous gamma globulin (1 g/kg) and an infusion of 20 U of platelets resulted in a transient rise in platelet count to 160,000/microliter, permitting surgery without bleeding. Danazol (800 mg/day) and conjugated estrogen (Premarin 25 mg/day) were begun to control the uterine bleeding. Intensive plasma exchange and i.v. IgG infusions were continued daily for 24 days, then twice weekly for several weeks. Platelet-bound IgG decreased almost 500% over the first 10 days of therapy, and platelets increased dramatically to 600,000/microliter after 3 weeks of therapy. The patient has remained amenorrheic with a normal platelet count for more than 24 months on daily danazol therapy and monthly infusions of i.v. IgG (0.4 g/kg/dose). | Aggressive combination therapy in the successful management of life-threatening intracranial hemorrhage in a patient with idiopathic thrombocytopenic purpura. We describe acute therapy for a 13-year-old female from Panama with chronic idiopathic thrombocytopenia purpura (ITP) refractory to steroids, splenectomy, vinca alkaloids, and azathioprine. She presented with neurologic deterioration from a posterior fossa intracranial hemorrhage (ICH). This followed a 3-month history of severe dysfunctional uterine bleeding, progressive from menarche, which had required multiple red cell transfusions. Steroid and vinblastine therapy and transfusion of 40 U of platelets failed to increase the platelet count above 10,000/microliter. Development of a second larger ICH (frontal) produced morbid increase in intracranial pressure that necessitated neurosurgical decompression. Plasma exchange and colloid repletion with intravenous gamma globulin (1 g/kg) and an infusion of 20 U of platelets resulted in a transient rise in platelet count to 160,000/microliter, permitting surgery without bleeding. Danazol (800 mg/day) and conjugated estrogen (Premarin 25 mg/day) were begun to control the uterine bleeding. Intensive plasma exchange and i.v. IgG infusions were continued daily for 24 days, then twice weekly for several weeks. Platelet-bound IgG decreased almost 500% over the first 10 days of therapy, and platelets increased dramatically to 600,000/microliter after 3 weeks of therapy. The patient has remained amenorrheic with a normal platelet count for more than 24 months on daily danazol therapy and monthly infusions of i.v. IgG (0.4 g/kg/dose). | 3,766,911 |
pubmed23n0125_13634 | Pediatric resident objectives for hematology/oncology. | Pediatric residents are exposed to a massive amount of scientific information and data from each pediatric subspecialty discipline. The residents may or may not know what is important and what is relevant from a general pediatrician's point of view. Educational objectives have been written by three pediatric hematologists/oncologists to guide the resident specifically as to the minimal expectations of knowledge and performance in this discipline. Other pediatric residency programs may wish to use the objectives as a starting point or an outline for their own objectives. | Pediatric resident objectives for hematology/oncology. Pediatric residents are exposed to a massive amount of scientific information and data from each pediatric subspecialty discipline. The residents may or may not know what is important and what is relevant from a general pediatrician's point of view. Educational objectives have been written by three pediatric hematologists/oncologists to guide the resident specifically as to the minimal expectations of knowledge and performance in this discipline. Other pediatric residency programs may wish to use the objectives as a starting point or an outline for their own objectives. | 3,766,912 |
pubmed23n0125_13635 | Treatment of localized neuroblastoma. | The treatment of neuroblastoma must take into account the prognosis as determined at the time of diagnosis. The stage is an important prognostic factor. In Evans's Stage I neuroblastoma, surgery is curative in itself and adjuvant treatment is not needed. In Evans' Stage II, it is our opinion that the presence of involved lymph nodes worsens the prognosis. Therefore, we suggest that patients with Stage II should be divided into Stage IIA (without lymph node involvement) and Stage IIB (with lymph node involvement). The treatment of Stage IIA patients may be the same as for Stage I patients, i.e., surgery alone. However, Stage IIB patients should be treated with intensive combined chemotherapy of short duration following surgery. | Treatment of localized neuroblastoma. The treatment of neuroblastoma must take into account the prognosis as determined at the time of diagnosis. The stage is an important prognostic factor. In Evans's Stage I neuroblastoma, surgery is curative in itself and adjuvant treatment is not needed. In Evans' Stage II, it is our opinion that the presence of involved lymph nodes worsens the prognosis. Therefore, we suggest that patients with Stage II should be divided into Stage IIA (without lymph node involvement) and Stage IIB (with lymph node involvement). The treatment of Stage IIA patients may be the same as for Stage I patients, i.e., surgery alone. However, Stage IIB patients should be treated with intensive combined chemotherapy of short duration following surgery. | 3,766,913 |
pubmed23n0125_13636 | Histologic criteria for the diagnosis of pityriasis rubra pilaris. | Even though the clinical appearance of pityriasis rubra pilaris is often distinctive enough to allow specific diagnosis of it, criteria for its histologic diagnosis have not yet been well defined. On the basis of a study of 55 cases of pityriasis rubra pilaris, we found the following histologic features helpful for diagnosis throughout most of the course of the disease: alternating orthokeratosis and parakeratosis in both vertical and horizontal directions; focal or confluent hypergranulosis; thick suprapapillary plates; broad rete ridges; narrow dermal papillae; and sparse superficial perivascular infiltration, mostly of lymphocytes. Plugging of follicular infundibula by cornified cells was seen only in biopsy specimens that came from lesions that were clinically indubitably follicular. Psoriasis has many histologic features in common with pityriasis rubra pilaris, but evolving and fully developed lesions of psoriasis have neutrophils in mounds of parakeratosis, thin rete ridges, thin suprapapillary plates, broad dermal papillae, and mixed inflammatory cell infiltrates of variable density. | Histologic criteria for the diagnosis of pityriasis rubra pilaris. Even though the clinical appearance of pityriasis rubra pilaris is often distinctive enough to allow specific diagnosis of it, criteria for its histologic diagnosis have not yet been well defined. On the basis of a study of 55 cases of pityriasis rubra pilaris, we found the following histologic features helpful for diagnosis throughout most of the course of the disease: alternating orthokeratosis and parakeratosis in both vertical and horizontal directions; focal or confluent hypergranulosis; thick suprapapillary plates; broad rete ridges; narrow dermal papillae; and sparse superficial perivascular infiltration, mostly of lymphocytes. Plugging of follicular infundibula by cornified cells was seen only in biopsy specimens that came from lesions that were clinically indubitably follicular. Psoriasis has many histologic features in common with pityriasis rubra pilaris, but evolving and fully developed lesions of psoriasis have neutrophils in mounds of parakeratosis, thin rete ridges, thin suprapapillary plates, broad dermal papillae, and mixed inflammatory cell infiltrates of variable density. | 3,766,917 |
pubmed23n0125_13637 | The fluorescence of melanocytic lesions. | The fluorescence of routine formalin-fixed sections of a variety of melanocytic lesions has been studied. The technique has value in distinguishing benign and atypical nevi from malignant melanomas. Forty-one of 48 melanomas exhibited positive fluorescence, whereas 15 of 86 of a variety of benign nevi were positive. The majority of the negative melanomas were Hutchinson's melanotic freckles, and the majority of the positive nevi were Spitz's nevi. Positive focal junctional fluorescence was a feature in up to half of the cases from established dysplastic nevus syndrome, but was noted only occasionally in sporadic dysplastic nevi. This distinguishes the dysplastic group from banal nevi and may be a marker of malignant potential. It is proposed that positive fluorescence is a marker of a melanocyte with an immature metabolic pathway. | The fluorescence of melanocytic lesions. The fluorescence of routine formalin-fixed sections of a variety of melanocytic lesions has been studied. The technique has value in distinguishing benign and atypical nevi from malignant melanomas. Forty-one of 48 melanomas exhibited positive fluorescence, whereas 15 of 86 of a variety of benign nevi were positive. The majority of the negative melanomas were Hutchinson's melanotic freckles, and the majority of the positive nevi were Spitz's nevi. Positive focal junctional fluorescence was a feature in up to half of the cases from established dysplastic nevus syndrome, but was noted only occasionally in sporadic dysplastic nevi. This distinguishes the dysplastic group from banal nevi and may be a marker of malignant potential. It is proposed that positive fluorescence is a marker of a melanocyte with an immature metabolic pathway. | 3,766,918 |
pubmed23n0125_13638 | Comparative ultrastructural study of generalized and localized granuloma annulare. | The dermal changes in four cases of generalized granuloma annulare (GGA) and in four of localized granuloma annulare (LGA) were studied ultrastructurally, and the findings in the two groups were compared. The basic alterations were similar in both types, and showed varied stages of development. The cellular infiltrate was composed mostly of histiocytes. Cell debris was found in all lesions. The degenerative changes affected the collagen fibers, the elastic fibers, and the cellular infiltrate. Of special interest are the following findings in GGA: First, masses of intercellular fibrin material were seen only in the lesions of GGA. Second, thick and multilayered basal lamina around capillaries was apparently more common in the generalized form. These changes may suggest that a more pronounced immune reaction is responsible for the development of the generalized form of the disease. | Comparative ultrastructural study of generalized and localized granuloma annulare. The dermal changes in four cases of generalized granuloma annulare (GGA) and in four of localized granuloma annulare (LGA) were studied ultrastructurally, and the findings in the two groups were compared. The basic alterations were similar in both types, and showed varied stages of development. The cellular infiltrate was composed mostly of histiocytes. Cell debris was found in all lesions. The degenerative changes affected the collagen fibers, the elastic fibers, and the cellular infiltrate. Of special interest are the following findings in GGA: First, masses of intercellular fibrin material were seen only in the lesions of GGA. Second, thick and multilayered basal lamina around capillaries was apparently more common in the generalized form. These changes may suggest that a more pronounced immune reaction is responsible for the development of the generalized form of the disease. | 3,766,919 |
pubmed23n0125_13639 | Penicillamine-induced elastosis perforans serpiginosa. Tip of the iceberg? | Elastosis perforans serpiginosa (EPS) is now a well-recognized potential complication of long-term penicillamine therapy. By itself, EPS appears to be a relatively innocuous cutaneous side effect of penicillamine. However, suspicion has been raised in recent literature that EPS may represent only a superficial manifestation of more serious penicillamine-induced systemic elastic tissue damage, particularly involving blood vessels. This is a report of a patient with Wilson's disease who was treated with penicillamine for 14 years. She developed EPS, and histologic examination of the skin revealed the characteristic penicillamine-induced "lumpy-bumpy" elastic fibers in the dermis. More important, nonlesional skin showed the same elastic fiber changes. Of greatest significance was the finding of identical elastic fiber alterations in an artery. | Penicillamine-induced elastosis perforans serpiginosa. Tip of the iceberg? Elastosis perforans serpiginosa (EPS) is now a well-recognized potential complication of long-term penicillamine therapy. By itself, EPS appears to be a relatively innocuous cutaneous side effect of penicillamine. However, suspicion has been raised in recent literature that EPS may represent only a superficial manifestation of more serious penicillamine-induced systemic elastic tissue damage, particularly involving blood vessels. This is a report of a patient with Wilson's disease who was treated with penicillamine for 14 years. She developed EPS, and histologic examination of the skin revealed the characteristic penicillamine-induced "lumpy-bumpy" elastic fibers in the dermis. More important, nonlesional skin showed the same elastic fiber changes. Of greatest significance was the finding of identical elastic fiber alterations in an artery. | 3,766,920 |
pubmed23n0125_13640 | Hypopigmented mycosis fungoides. Speculations about the mechanism of hypopigmentation. | Hypopigmented, nonatrophic macules are an unusual manifestation of lesions of early mycosis fungoides. Presented herein is a patient with such lesions and an unusual mask-like hypopigmentation of the face. We review both the clinical and microscopic findings in this patient and in previously reported patients with hypopigmented lesions of mycosis fungoides. Finally, speculation is made as to the mechanisms of the hypopigmentation. | Hypopigmented mycosis fungoides. Speculations about the mechanism of hypopigmentation. Hypopigmented, nonatrophic macules are an unusual manifestation of lesions of early mycosis fungoides. Presented herein is a patient with such lesions and an unusual mask-like hypopigmentation of the face. We review both the clinical and microscopic findings in this patient and in previously reported patients with hypopigmented lesions of mycosis fungoides. Finally, speculation is made as to the mechanisms of the hypopigmentation. | 3,766,921 |
pubmed23n0125_13641 | The enigma of juvenile plantar dermatosis. | Juvenile plantar dermatosis, which is sometimes accompanied by palmar changes, is a chronic condition. It is characterized by shiny redness, anhidrosis, and fissuring of the skin. Microscopically, focal inflammatory changes at the junctions of sweat-gland ducts and acrosyringia are seen. The pathogenesis is unknown. We surmise that individual susceptibility, bacterial colonization, and the use of man-made leather substitutes and fibers are the main causative factors. | The enigma of juvenile plantar dermatosis. Juvenile plantar dermatosis, which is sometimes accompanied by palmar changes, is a chronic condition. It is characterized by shiny redness, anhidrosis, and fissuring of the skin. Microscopically, focal inflammatory changes at the junctions of sweat-gland ducts and acrosyringia are seen. The pathogenesis is unknown. We surmise that individual susceptibility, bacterial colonization, and the use of man-made leather substitutes and fibers are the main causative factors. | 3,766,922 |
pubmed23n0125_13642 | Nasal glioma. A rarity. | Nasal glioma is a rare congenital malformation which most often presents to the otorhinolaryngologist or neurosurgeon. However, occasional cases of the extranasal variant are referred to a dermatologist as an unusual subcutaneous mass over the bridge of the nose, most often in an infant. These lesions may not be recognised clinically, leading to dangerous mismanagement, and may be misinterpreted by pathologists as a neoplasm, either benign or malignant. Three cases are described here with the purpose of focussing the attention of workers in dermatology on this unusual developmental abnormality. | Nasal glioma. A rarity. Nasal glioma is a rare congenital malformation which most often presents to the otorhinolaryngologist or neurosurgeon. However, occasional cases of the extranasal variant are referred to a dermatologist as an unusual subcutaneous mass over the bridge of the nose, most often in an infant. These lesions may not be recognised clinically, leading to dangerous mismanagement, and may be misinterpreted by pathologists as a neoplasm, either benign or malignant. Three cases are described here with the purpose of focussing the attention of workers in dermatology on this unusual developmental abnormality. | 3,766,923 |
pubmed23n0125_13643 | Paranasal sinus pathology in allergic and non-allergic respiratory tract diseases. | Two hundred and seventy patients with asthma and/or rhinitis (162 or 60% allergic, 108 or 40% non-allergic) were studied for sinus pathology by means of standard X-rays and tomograms. Sinus pathology was defined as abnormal sinus X-rays, either on standard or tomography. Fifty-four percent of the X-rays were classified as abnormal based on mucosal thickening, loss of translucency of the cavities of polyps. Asthma was significantly more often associated with sinus X-ray abnormalities (65.1%) than rhinitis and/or chronic cough (44.4%). Loss of translucency of the cavities is more frequent in children, whereas mucosa thickening becomes more frequent with progressing age. Since in this prospective study the taking of X-rays of the sinuses was not dependent on or related to temporarily occurring symptoms which could be attributed to acute sinusitis, the presence of sinus abnormalities on X-rays can be considered as an indicator of the chronicity of airways diseases and might provide an indication for prophylactic therapy of the associated airway disease in a continuous way. The importance of sinus tomograms is stressed, since only 32.5% of the patients with mucosa thickening could be detected on standard X-rays. | Paranasal sinus pathology in allergic and non-allergic respiratory tract diseases. Two hundred and seventy patients with asthma and/or rhinitis (162 or 60% allergic, 108 or 40% non-allergic) were studied for sinus pathology by means of standard X-rays and tomograms. Sinus pathology was defined as abnormal sinus X-rays, either on standard or tomography. Fifty-four percent of the X-rays were classified as abnormal based on mucosal thickening, loss of translucency of the cavities of polyps. Asthma was significantly more often associated with sinus X-ray abnormalities (65.1%) than rhinitis and/or chronic cough (44.4%). Loss of translucency of the cavities is more frequent in children, whereas mucosa thickening becomes more frequent with progressing age. Since in this prospective study the taking of X-rays of the sinuses was not dependent on or related to temporarily occurring symptoms which could be attributed to acute sinusitis, the presence of sinus abnormalities on X-rays can be considered as an indicator of the chronicity of airways diseases and might provide an indication for prophylactic therapy of the associated airway disease in a continuous way. The importance of sinus tomograms is stressed, since only 32.5% of the patients with mucosa thickening could be detected on standard X-rays. | 3,766,930 |
pubmed23n0125_13644 | Comparative evaluation of the multiple RAST and discrete RAST for inhalant allergy. | With the conventional, discrete RAST various tests are required to detect IgE of different specificities in the same serum. To overcome this problem and to reduce the costs, a multiple RAST with seven different mixtures was compared with the individual mixture constituents and with the 12 individual allergens currently in use in our department. One grass pollen mixture (gx3), two weed pollen mixtures (wx3, wx4), two tree pollen mixtures (tx5, tx6), one mould mixture (mx1) and one epithelial mixture (ex1) were used. A mixture of mites was not evaluated as there is only one important pathogenic organism in our regions (Dermatophagoides pteronyssinus or House dust mite). For grasses the gx3 mixture offered no advantage over the discrete RAST. The weed mixture wx3 was more sensitive than the most common discrete RAST's but at the cost of specificity. The wx4 mixture should not be used because the specificity is too low. The tree mixtures were not significantly more sensitive than the most common individual tree allergens, and were less specific. Mould mixtures should not be used because there is little cross-reactivity between the individual allergens, thus using a mixture would necessitate the subsequent determination of individual allergens, and the number of tests and the cost would be even higher. Neither should a mixture be used for epithelia because one wants to detect allergies to individual allergens. Moreover, the sensitivity of the epithelial mixture was too low. In general, we suggest the use of a limited panel of discrete RAST's instead of mixtures. | Comparative evaluation of the multiple RAST and discrete RAST for inhalant allergy. With the conventional, discrete RAST various tests are required to detect IgE of different specificities in the same serum. To overcome this problem and to reduce the costs, a multiple RAST with seven different mixtures was compared with the individual mixture constituents and with the 12 individual allergens currently in use in our department. One grass pollen mixture (gx3), two weed pollen mixtures (wx3, wx4), two tree pollen mixtures (tx5, tx6), one mould mixture (mx1) and one epithelial mixture (ex1) were used. A mixture of mites was not evaluated as there is only one important pathogenic organism in our regions (Dermatophagoides pteronyssinus or House dust mite). For grasses the gx3 mixture offered no advantage over the discrete RAST. The weed mixture wx3 was more sensitive than the most common discrete RAST's but at the cost of specificity. The wx4 mixture should not be used because the specificity is too low. The tree mixtures were not significantly more sensitive than the most common individual tree allergens, and were less specific. Mould mixtures should not be used because there is little cross-reactivity between the individual allergens, thus using a mixture would necessitate the subsequent determination of individual allergens, and the number of tests and the cost would be even higher. Neither should a mixture be used for epithelia because one wants to detect allergies to individual allergens. Moreover, the sensitivity of the epithelial mixture was too low. In general, we suggest the use of a limited panel of discrete RAST's instead of mixtures. | 3,766,931 |
pubmed23n0125_13645 | Long-term changes in inhalant allergy in asthmatic children. | In a representative cohort of 55 asthmatic schoolchildren the progress of the allergy per se was followed up prospectively for 8 years. Judged by clinical data, skin prick tests and RAST, a large majority of the children retained their allergies to pollen and animal danders. These allergens were predominant, whereas allergy to mites and moulds was less frequent. Serum IgE levels showed a strong tendency to remain high. Positive RAST and skin prick tests were also found in a substantial number of children with normal serum IgE concentrations. | Long-term changes in inhalant allergy in asthmatic children. In a representative cohort of 55 asthmatic schoolchildren the progress of the allergy per se was followed up prospectively for 8 years. Judged by clinical data, skin prick tests and RAST, a large majority of the children retained their allergies to pollen and animal danders. These allergens were predominant, whereas allergy to mites and moulds was less frequent. Serum IgE levels showed a strong tendency to remain high. Positive RAST and skin prick tests were also found in a substantial number of children with normal serum IgE concentrations. | 3,766,932 |
pubmed23n0125_13646 | Quantitative determination of antiviral nucleoside analog in DNA. | A technique for the analysis of the amount of an antiviral nucleoside analog incorporated into DNA, utilizing enzymatic digestion of DNA, followed by high-performance liquid chromatography is described. The cells or tissue samples were treated with perchloric acid to inactivate the nucleases, then digested with pronase in the presence of EDTA. DNA was purified by CsCl centrifugation followed by Sephadex chromatography and treatment with deoxyribonuclease 1 and venom phosphodiesterase. The deoxyribonucleoside monophosphates and the monophosphate of the nucleoside analog liberated from DNA were separated and quantitated by HPLC analysis and measurement of radioactivity. This assay is more sensitive, specific, and precise than the determination of DNA density shift. It is also applicable for nucleoside analogs which do not change the density of DNA either because of their structure or their very small degree of incorporation. | Quantitative determination of antiviral nucleoside analog in DNA. A technique for the analysis of the amount of an antiviral nucleoside analog incorporated into DNA, utilizing enzymatic digestion of DNA, followed by high-performance liquid chromatography is described. The cells or tissue samples were treated with perchloric acid to inactivate the nucleases, then digested with pronase in the presence of EDTA. DNA was purified by CsCl centrifugation followed by Sephadex chromatography and treatment with deoxyribonuclease 1 and venom phosphodiesterase. The deoxyribonucleoside monophosphates and the monophosphate of the nucleoside analog liberated from DNA were separated and quantitated by HPLC analysis and measurement of radioactivity. This assay is more sensitive, specific, and precise than the determination of DNA density shift. It is also applicable for nucleoside analogs which do not change the density of DNA either because of their structure or their very small degree of incorporation. | 3,766,933 |
pubmed23n0125_13647 | The use of spun columns in the purification of double-stranded cDNA appropriate for tailing. | A reliable estimation of the efficiency of adding homopolymeric tails to double-stranded cDNA molecules by terminal deoxynucleotidyl transferase is extremely important in the construction of cDNA libraries. Appreciable differences in transformation efficiency result when the homopolymer tails to be annealed are of inadequate length. We report here that the use of a Sephadex G-50 spun column to remove oligo(dT)12-18 frequently coprecipitating in ethanol with the cDNA results in a more dependable estimation of tail lengths. | The use of spun columns in the purification of double-stranded cDNA appropriate for tailing. A reliable estimation of the efficiency of adding homopolymeric tails to double-stranded cDNA molecules by terminal deoxynucleotidyl transferase is extremely important in the construction of cDNA libraries. Appreciable differences in transformation efficiency result when the homopolymer tails to be annealed are of inadequate length. We report here that the use of a Sephadex G-50 spun column to remove oligo(dT)12-18 frequently coprecipitating in ethanol with the cDNA results in a more dependable estimation of tail lengths. | 3,766,934 |
pubmed23n0125_13648 | A dot-blot assay for heparin-binding proteins. | A method for the detection and quantitation of picomole amounts of heparin-binding proteins is described. Proteins are first spotted on nitrocellulose and then incubated with 125I-heparin. Binding of heparin to the proteins is detected by radioautography and quantitated by scanning densitometry; proteins are quantitated by densitometric analysis of the amido black stained nitrocellulose. Heparin-binding was time-dependent and sensitive to the presence of metal ions, urea, and detergents (anionic, nonionic, and zwitterionic). The divalent cations Ca2+ and Mg2+ and the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate increased heparin binding whereas NaCl, urea, sodium dodecylsulfate, and La3+ decreased binding. This assay is applicable to the identification and characterization of a variety of heparin-binding proteins. | A dot-blot assay for heparin-binding proteins. A method for the detection and quantitation of picomole amounts of heparin-binding proteins is described. Proteins are first spotted on nitrocellulose and then incubated with 125I-heparin. Binding of heparin to the proteins is detected by radioautography and quantitated by scanning densitometry; proteins are quantitated by densitometric analysis of the amido black stained nitrocellulose. Heparin-binding was time-dependent and sensitive to the presence of metal ions, urea, and detergents (anionic, nonionic, and zwitterionic). The divalent cations Ca2+ and Mg2+ and the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate increased heparin binding whereas NaCl, urea, sodium dodecylsulfate, and La3+ decreased binding. This assay is applicable to the identification and characterization of a variety of heparin-binding proteins. | 3,766,935 |
pubmed23n0125_13649 | High-performance liquid chromatography analysis of the thiobarbituric acid adducts of malonaldehyde and trans,trans-muconaldehyde. | A reversed-phase HPLC method is described for the separation and analysis of the thiobarbituric acid (TBA) adducts of the reactive aldehydes muconaldehyde (MUC) and malonaldehyde (MDA). The TBA adduct of malonaldehyde was synthesized, purified, and its structure elucidated, for use as standard in quantitative HPLC studies. A detection limit of 1 X 10(-14) mol was achieved for the MUC:TBA and MDA:TBA adducts using the double monochromator fluorometric detector, 7 X 10(-13) mol was the detection limit using a variable wavelength uv-visible detector. Direct on-line identification of the eluting aldehyde:TBA adducts was achieved by the use of a diode-array uv-visible detector. The chromatographic behavior of the adducts under different mobile phase conditions was also examined. This HPLC methodology was used for the identification of muconaldehyde as a product of benzene oxidation in a hydroxyl radical generating system. | High-performance liquid chromatography analysis of the thiobarbituric acid adducts of malonaldehyde and trans,trans-muconaldehyde. A reversed-phase HPLC method is described for the separation and analysis of the thiobarbituric acid (TBA) adducts of the reactive aldehydes muconaldehyde (MUC) and malonaldehyde (MDA). The TBA adduct of malonaldehyde was synthesized, purified, and its structure elucidated, for use as standard in quantitative HPLC studies. A detection limit of 1 X 10(-14) mol was achieved for the MUC:TBA and MDA:TBA adducts using the double monochromator fluorometric detector, 7 X 10(-13) mol was the detection limit using a variable wavelength uv-visible detector. Direct on-line identification of the eluting aldehyde:TBA adducts was achieved by the use of a diode-array uv-visible detector. The chromatographic behavior of the adducts under different mobile phase conditions was also examined. This HPLC methodology was used for the identification of muconaldehyde as a product of benzene oxidation in a hydroxyl radical generating system. | 3,766,936 |
pubmed23n0125_13650 | Stability of o-phthalaldehyde-derived isoindoles. | The degradation of 1-alkylthio-2-alkylisoindoles, generated in situ by reaction of o-phthalaldehyde with primary alkylamines in the presence of selected thiols, was studied under conditions typical of those used in precolumn derivatization applications. In all cases, loss of isoindole followed pseudo first order kinetics and was strongly accelerated by excess o-phthalaldehyde in the reaction mixture. Reactivity toward o-phthalaldehyde was found to be influenced by a number of experimental parameters including thiol structure, thiol concentration, amine structure, solvent composition, and pH. In the absence of excess o-phthalaldehyde, the stability of all derivatives examined was excellent. These results provide a basis for optimization of reaction conditions for maximum derivative stability. | Stability of o-phthalaldehyde-derived isoindoles. The degradation of 1-alkylthio-2-alkylisoindoles, generated in situ by reaction of o-phthalaldehyde with primary alkylamines in the presence of selected thiols, was studied under conditions typical of those used in precolumn derivatization applications. In all cases, loss of isoindole followed pseudo first order kinetics and was strongly accelerated by excess o-phthalaldehyde in the reaction mixture. Reactivity toward o-phthalaldehyde was found to be influenced by a number of experimental parameters including thiol structure, thiol concentration, amine structure, solvent composition, and pH. In the absence of excess o-phthalaldehyde, the stability of all derivatives examined was excellent. These results provide a basis for optimization of reaction conditions for maximum derivative stability. | 3,766,937 |
pubmed23n0125_13651 | A high-performance liquid chromatographic method for the determination of dolichyl phosphates in tissues. | A sensitive high-performance liquid chromatographic method has been developed for the determination of dolichyl phosphate homologs in tissues. Dolichyl phosphates were methylated and derivatized with 3-(9-anthryl)-diazo-2-propene to yield methyl 3-(9-anthryl)-2-propenyl dolichyl phosphates. As an internal standard, 2,2-didecaprenylethyl phosphate was used. The derivatives were separated on a reversed-phase column using isocratic elution, and detected fluorometrically. The method with fluorescence detection was sufficiently sensitive to measure the concentration of dolichyl phosphate homologs in tissues. | A high-performance liquid chromatographic method for the determination of dolichyl phosphates in tissues. A sensitive high-performance liquid chromatographic method has been developed for the determination of dolichyl phosphate homologs in tissues. Dolichyl phosphates were methylated and derivatized with 3-(9-anthryl)-diazo-2-propene to yield methyl 3-(9-anthryl)-2-propenyl dolichyl phosphates. As an internal standard, 2,2-didecaprenylethyl phosphate was used. The derivatives were separated on a reversed-phase column using isocratic elution, and detected fluorometrically. The method with fluorescence detection was sufficiently sensitive to measure the concentration of dolichyl phosphate homologs in tissues. | 3,766,938 |
pubmed23n0125_13652 | A fluorometric method for the determination of triglycerides in nanomolar quantities. | A fluorometric assay for triglycerides in nanomole quantities is described. Glycerol is liberated from triglycerides with lipase from Chromobacter viscosum, then converted by glycerol kinase to glycerol-3-phosphate, which is oxidized by glycerol-3-phosphate oxidase, producing H2O2. The H2O2 ultimately forms a peroxidase-catalyzed fluorogen with p-hydroxyphenylacetic acid. The excitation and emission wavelengths of the fluorogen are 325 and 415 nm, respectively. The assay is linear in the range 0.05-35 nmol of triglycerides using triolein as standard. | A fluorometric method for the determination of triglycerides in nanomolar quantities. A fluorometric assay for triglycerides in nanomole quantities is described. Glycerol is liberated from triglycerides with lipase from Chromobacter viscosum, then converted by glycerol kinase to glycerol-3-phosphate, which is oxidized by glycerol-3-phosphate oxidase, producing H2O2. The H2O2 ultimately forms a peroxidase-catalyzed fluorogen with p-hydroxyphenylacetic acid. The excitation and emission wavelengths of the fluorogen are 325 and 415 nm, respectively. The assay is linear in the range 0.05-35 nmol of triglycerides using triolein as standard. | 3,766,939 |
pubmed23n0125_13653 | One-step procedure for the determination of thymosin beta 4 in small tissue samples and its separation from other thymosin beta 4-like peptides by high-pressure liquid chromatography. | Thymosin beta 4 has been determined by a simple and fast one-step procedure in different tissues of rats. The tissues (1 to 40 mg) were disintegrated and deproteinized by homogenization in perchloric acid. After neutralization by potassium hydroxide the supernatant solution was used for determining thymosin beta 4 by reverse-phase HPLC without further manipulations. Not only does this procedure avoid artificial proteolysis as effectively as extraction of tissues by guanidinium chloride or boiling buffer, but it offers two further advantages. First, no additional steps--as for example desalting--are necessary prior to HPLC and thus the risk of losing thymosin beta 4 is eliminated. Using this procedure thymosin beta 4 is recovered quantitatively. The method is linear over the range 0.04 to 1.13 nmol and thymosin beta 4 is well separated from other thymosin beta 4-like peptides known to be present in mammals; i.e., thymosin beta Ala4, thymosin beta 9, thymosin beta 10, and thymosin beta Arg10. Second, the acid-insoluble pellet of the same extract can be used to determine the DNA content of the sample. Thus it is possible to relate thymosin beta 4 to DNA, which then allows comparing cells of different tissues and cell lines to one another. This procedure is also applicable to small peptides soluble in perchloric acid. | One-step procedure for the determination of thymosin beta 4 in small tissue samples and its separation from other thymosin beta 4-like peptides by high-pressure liquid chromatography. Thymosin beta 4 has been determined by a simple and fast one-step procedure in different tissues of rats. The tissues (1 to 40 mg) were disintegrated and deproteinized by homogenization in perchloric acid. After neutralization by potassium hydroxide the supernatant solution was used for determining thymosin beta 4 by reverse-phase HPLC without further manipulations. Not only does this procedure avoid artificial proteolysis as effectively as extraction of tissues by guanidinium chloride or boiling buffer, but it offers two further advantages. First, no additional steps--as for example desalting--are necessary prior to HPLC and thus the risk of losing thymosin beta 4 is eliminated. Using this procedure thymosin beta 4 is recovered quantitatively. The method is linear over the range 0.04 to 1.13 nmol and thymosin beta 4 is well separated from other thymosin beta 4-like peptides known to be present in mammals; i.e., thymosin beta Ala4, thymosin beta 9, thymosin beta 10, and thymosin beta Arg10. Second, the acid-insoluble pellet of the same extract can be used to determine the DNA content of the sample. Thus it is possible to relate thymosin beta 4 to DNA, which then allows comparing cells of different tissues and cell lines to one another. This procedure is also applicable to small peptides soluble in perchloric acid. | 3,766,940 |
pubmed23n0125_13654 | A micromethod for the determination of iron and total iron-binding capacity in intraocular fluids and plasma using electrothermal atomic absorption spectroscopy. | A new micromethod (requiring only 30 microliters of sample) for the determination of iron (Fe) concentration and total iron-binding capacity (TIBC) in intraocular fluids (IOFs; aqueous and vitreous humors) and plasma has been developed using electrothermal atomic absorption spectrophotometry. Values for Fe concentration in the IOFs were similar between species and were generally higher in the vitreous (0.031-0.060 mg/liter) than aqueous (0.014-0.038 mg/liter) humors obtained from the same eye. Plasma Fe concentration and TIBC fell within established limits for all species studied. The iron binding capacity of the aqueous and vitreous humors from the same species is similar. While the TIBC of the aqueous humor is saturated to the same extent as plasma, vitreous TIBC is more highly saturated in all species. Using this method considerably less species variation in IOF Fe concentration was found as compared to previous studies. Some possible methodological reasons for this difference are discussed. | A micromethod for the determination of iron and total iron-binding capacity in intraocular fluids and plasma using electrothermal atomic absorption spectroscopy. A new micromethod (requiring only 30 microliters of sample) for the determination of iron (Fe) concentration and total iron-binding capacity (TIBC) in intraocular fluids (IOFs; aqueous and vitreous humors) and plasma has been developed using electrothermal atomic absorption spectrophotometry. Values for Fe concentration in the IOFs were similar between species and were generally higher in the vitreous (0.031-0.060 mg/liter) than aqueous (0.014-0.038 mg/liter) humors obtained from the same eye. Plasma Fe concentration and TIBC fell within established limits for all species studied. The iron binding capacity of the aqueous and vitreous humors from the same species is similar. While the TIBC of the aqueous humor is saturated to the same extent as plasma, vitreous TIBC is more highly saturated in all species. Using this method considerably less species variation in IOF Fe concentration was found as compared to previous studies. Some possible methodological reasons for this difference are discussed. | 3,766,941 |
pubmed23n0125_13655 | Rapid preparation of bacteriophage DNA for sequence analysis in sets of 96 clones, using filtration. | A method is described for the preparation of single-stranded DNA from clones in bacteriophage M13 vectors. This procedure allows multiples of 96 clones to be processed at once, utilizing filtration to remove host cells and simplifying the treatment of bacteriophage pellets. The DNA produced can be used for sequencing of mutagenesis. | Rapid preparation of bacteriophage DNA for sequence analysis in sets of 96 clones, using filtration. A method is described for the preparation of single-stranded DNA from clones in bacteriophage M13 vectors. This procedure allows multiples of 96 clones to be processed at once, utilizing filtration to remove host cells and simplifying the treatment of bacteriophage pellets. The DNA produced can be used for sequencing of mutagenesis. | 3,766,942 |
pubmed23n0125_13656 | Fast atom bombardment combined with tandem mass spectrometry for the study of dinucleotides. | A study of mono- and dinucleotides by utilizing negative ion fast atom bombardment (FAB), metastable decomposition of (M-H)- species, and collisionally activated decomposition (CAD) of (M-H)- species is reported. Data were obtained for several complete series containing the standard nucleosides (guanosine, adenosine, cytidine, thymidine, and uridine): the 3'- and 5'-monophosphate mononucleotide series for both ribo- and 2'-deoxyribomononucleotides, all possible combinations for the 3'(-)----5'-ribodinucleotides, and all possible combinations of the 3'(-)----5',2'-deoxyribodinucleotides. The metastable and CAD spectra provide more information than the FAB mass spectra. The (M-H)- ions of all dinucleotides decompose either as metastable ions or upon collisional activation to eliminate BH (B = base) preferentially from the 3'- rather than the 5'-terminus. Isomeric dinucleotides can be distinguished on the basis of this fragmentation. To establish the identity of the base at the 5'-terminus, collisional activation is preferred. By comparing relative abundances of BH elimination observed, the inherent basicities of the nucleoside base anions can be inferred to be C- greater than A-, T-, greater than G-. | Fast atom bombardment combined with tandem mass spectrometry for the study of dinucleotides. A study of mono- and dinucleotides by utilizing negative ion fast atom bombardment (FAB), metastable decomposition of (M-H)- species, and collisionally activated decomposition (CAD) of (M-H)- species is reported. Data were obtained for several complete series containing the standard nucleosides (guanosine, adenosine, cytidine, thymidine, and uridine): the 3'- and 5'-monophosphate mononucleotide series for both ribo- and 2'-deoxyribomononucleotides, all possible combinations for the 3'(-)----5'-ribodinucleotides, and all possible combinations of the 3'(-)----5',2'-deoxyribodinucleotides. The metastable and CAD spectra provide more information than the FAB mass spectra. The (M-H)- ions of all dinucleotides decompose either as metastable ions or upon collisional activation to eliminate BH (B = base) preferentially from the 3'- rather than the 5'-terminus. Isomeric dinucleotides can be distinguished on the basis of this fragmentation. To establish the identity of the base at the 5'-terminus, collisional activation is preferred. By comparing relative abundances of BH elimination observed, the inherent basicities of the nucleoside base anions can be inferred to be C- greater than A-, T-, greater than G-. | 3,766,943 |
pubmed23n0125_13657 | Alternating current voltammetric determination of DNA concentrations at a microgram per liter level. | Alternating current voltammetry is used as a fast and highly sensitive method of DNA detection, at a microgram per liter level. The method is based on the measurement of adsorption effects of denatured DNA at the hanging mercury drop electrode. The proposed procedure consists of thermal denaturation of DNA, which is followed by electrochemical detection of denatured DNA. A sharp adsorption peak of denatured DNA, at the potential of -1.4 V, is measured in 0.3 mol/liter NaCl and 0.03 mol/liter NaHCO3 (pH about 9) after an accumulation of DNA at the electrode surface. To enhance the sensitivity, the solution is stirred during adsorption. The influence of proteins, a polysaccharide, and RNA on the DNA determination was also studied. | Alternating current voltammetric determination of DNA concentrations at a microgram per liter level. Alternating current voltammetry is used as a fast and highly sensitive method of DNA detection, at a microgram per liter level. The method is based on the measurement of adsorption effects of denatured DNA at the hanging mercury drop electrode. The proposed procedure consists of thermal denaturation of DNA, which is followed by electrochemical detection of denatured DNA. A sharp adsorption peak of denatured DNA, at the potential of -1.4 V, is measured in 0.3 mol/liter NaCl and 0.03 mol/liter NaHCO3 (pH about 9) after an accumulation of DNA at the electrode surface. To enhance the sensitivity, the solution is stirred during adsorption. The influence of proteins, a polysaccharide, and RNA on the DNA determination was also studied. | 3,766,945 |
pubmed23n0125_13658 | Poly(ADP-ribosyl)ation studies in situ. | We have studied the poly(ADP-ribosyl)ation of nuclear proteins in situ by examining the incorporation of [3H]NAD-derived ADP-ribose into polymers. We have devised a way to deliver [3H]NAD to cells growing in vitro, and we have determined the kinetics of uptake and incorporation into nuclear proteins using this delivery system. Incorporation into the histone fraction, known acceptors of poly(ADP-ribose), was examined and shown to be sensitive to the poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide. Polyacrylamide gel electrophoresis of 3H-labeled proteins revealed radioactivity associated with known poly(ADP-ribose)-accepting proteins such as poly(ADP-ribose) polymerase and histones. These results were confirmed when we immunoreacted gel-separated proteins with anti-(ADP-ribose) generated in our laboratory. | Poly(ADP-ribosyl)ation studies in situ. We have studied the poly(ADP-ribosyl)ation of nuclear proteins in situ by examining the incorporation of [3H]NAD-derived ADP-ribose into polymers. We have devised a way to deliver [3H]NAD to cells growing in vitro, and we have determined the kinetics of uptake and incorporation into nuclear proteins using this delivery system. Incorporation into the histone fraction, known acceptors of poly(ADP-ribose), was examined and shown to be sensitive to the poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide. Polyacrylamide gel electrophoresis of 3H-labeled proteins revealed radioactivity associated with known poly(ADP-ribose)-accepting proteins such as poly(ADP-ribose) polymerase and histones. These results were confirmed when we immunoreacted gel-separated proteins with anti-(ADP-ribose) generated in our laboratory. | 3,766,944 |
pubmed23n0125_13659 | Specific isolation of surface glycoproteins from intact cells by biotinylated concanavalin A and immobilized streptavidin. | An indirect affinity chromatography procedure utilizing biotinylated lectins and designed for the specific isolation of surface glycoproteins is described. The method is illustrated with intact acute leukemic lymphoblastic cells (ALL cells) with biotin-epsilon-aminocaproyl-concanavalin A (biocap-Con A) and streptavidin-Sepharose 4B. Biocap-Con A, containing on average 27 biotin residues per tetrameric lectin molecule, is used to isolate Con A-binding glycoproteins from the surface of [35S]methionine-radiolabeled intact cells. The biocap-Con A/glycoprotein complexes, after solubilization in detergent, are retrieved on immobilized streptavidin. The surface glycoproteins isolated from intact ALL cells by this method are subjected to two-dimensional gel electrophoresis and detected by autoradiography. More than fifty Con A-binding glycoproteins can be separated from the ALL cells. These glycoproteins retrievable from the cell surface were compared to those retrieved by the indirect affinity chromatography procedure from isolated plasma membrane fractions. Certain groups of glycoproteins present in the fraction isolated from intact cells were not detected in that from the plasma membrane preparations. The advantage of using the biocap-con A/streptavidin system with intact cells rather than isolated plasma membranes for the detection of surface glycoproteins is discussed. | Specific isolation of surface glycoproteins from intact cells by biotinylated concanavalin A and immobilized streptavidin. An indirect affinity chromatography procedure utilizing biotinylated lectins and designed for the specific isolation of surface glycoproteins is described. The method is illustrated with intact acute leukemic lymphoblastic cells (ALL cells) with biotin-epsilon-aminocaproyl-concanavalin A (biocap-Con A) and streptavidin-Sepharose 4B. Biocap-Con A, containing on average 27 biotin residues per tetrameric lectin molecule, is used to isolate Con A-binding glycoproteins from the surface of [35S]methionine-radiolabeled intact cells. The biocap-Con A/glycoprotein complexes, after solubilization in detergent, are retrieved on immobilized streptavidin. The surface glycoproteins isolated from intact ALL cells by this method are subjected to two-dimensional gel electrophoresis and detected by autoradiography. More than fifty Con A-binding glycoproteins can be separated from the ALL cells. These glycoproteins retrievable from the cell surface were compared to those retrieved by the indirect affinity chromatography procedure from isolated plasma membrane fractions. Certain groups of glycoproteins present in the fraction isolated from intact cells were not detected in that from the plasma membrane preparations. The advantage of using the biocap-con A/streptavidin system with intact cells rather than isolated plasma membranes for the detection of surface glycoproteins is discussed. | 3,766,946 |
pubmed23n0125_13660 | A new highly selective physicochemical assay to measure NAD+ in intact cells. | A simple, fast, and highly specific chromatographic method for measuring the content of NAD+ in intact cells has been developed. This procedure involves the separation of NAD+ from the bulk of acid-soluble nucleosides, nucleotides, and other pyridine containing molecules by affinity chromatography on dihydroxyboronyl-Bio-Rex. The boronate purified preparations were utilized for the quantification of NAD+ by strong anion exchange high-pressure liquid chromatography under isocratic conditions using a low salt buffer system. The overall recovery of the method exceeded 80%. This new method was applied to determine the extent of NAD+ consumption in intact hepatocytes following treatment with two different DNA damaging agents. A major advantage of this method is that it allows for the simultaneous determination of poly(ADP-ribose) in the acid-insoluble fraction of the same sample. | A new highly selective physicochemical assay to measure NAD+ in intact cells. A simple, fast, and highly specific chromatographic method for measuring the content of NAD+ in intact cells has been developed. This procedure involves the separation of NAD+ from the bulk of acid-soluble nucleosides, nucleotides, and other pyridine containing molecules by affinity chromatography on dihydroxyboronyl-Bio-Rex. The boronate purified preparations were utilized for the quantification of NAD+ by strong anion exchange high-pressure liquid chromatography under isocratic conditions using a low salt buffer system. The overall recovery of the method exceeded 80%. This new method was applied to determine the extent of NAD+ consumption in intact hepatocytes following treatment with two different DNA damaging agents. A major advantage of this method is that it allows for the simultaneous determination of poly(ADP-ribose) in the acid-insoluble fraction of the same sample. | 3,766,947 |
pubmed23n0125_13661 | The titration of the active centers of cellobiohydrolase from Trichoderma reesei. | A novel approach has been developed for the titration of enzyme active centers and for the determination of the molecular activity of enzymes. It is based on the simultaneous use of a nonspecific chromogenic substrate and a specific ligand (a substrate or an inhibitor), the latter being tightly bound with the enzyme's active center. The approach is demonstrated using the titration (that is, the determination of the molar concentration of the enzyme active centers) of purified cellobiohydrolase I (CBH I) (EC 3.2.1.91) of the fungus Trichoderma reesei. p-Nitrophenyl-beta-D-lactoside was used as a reference substrate (Km = 0.5 mM), and cellobiose and CM-cellulose as specific ligands. The molecular weight of CBH I as it was determined by the titration with cellobiose was 42,000 +/- 3,000. The inhibition constant by cellobiose was (6 +/- 1) X 10(-6) M. The value of the catalytic constant for the hydrolysis of p-nitrophenyl-beta-D-lactoside calculated from the titration data was equal to 0.063 s-1. CM-cellulose turned out to be more efficient titration agent for cellobiohydrolase than cellobiose, and might be used for the titration of the enzyme in concentrations of the latter of 0.008-0.02 mg/ml. The titration data showed that the inhibition constant of CM-cellulose toward CBH I was equal to (1.0 +/- 0.2) X 10(-7) M. | The titration of the active centers of cellobiohydrolase from Trichoderma reesei. A novel approach has been developed for the titration of enzyme active centers and for the determination of the molecular activity of enzymes. It is based on the simultaneous use of a nonspecific chromogenic substrate and a specific ligand (a substrate or an inhibitor), the latter being tightly bound with the enzyme's active center. The approach is demonstrated using the titration (that is, the determination of the molar concentration of the enzyme active centers) of purified cellobiohydrolase I (CBH I) (EC 3.2.1.91) of the fungus Trichoderma reesei. p-Nitrophenyl-beta-D-lactoside was used as a reference substrate (Km = 0.5 mM), and cellobiose and CM-cellulose as specific ligands. The molecular weight of CBH I as it was determined by the titration with cellobiose was 42,000 +/- 3,000. The inhibition constant by cellobiose was (6 +/- 1) X 10(-6) M. The value of the catalytic constant for the hydrolysis of p-nitrophenyl-beta-D-lactoside calculated from the titration data was equal to 0.063 s-1. CM-cellulose turned out to be more efficient titration agent for cellobiohydrolase than cellobiose, and might be used for the titration of the enzyme in concentrations of the latter of 0.008-0.02 mg/ml. The titration data showed that the inhibition constant of CM-cellulose toward CBH I was equal to (1.0 +/- 0.2) X 10(-7) M. | 3,766,948 |
pubmed23n0125_13662 | Assay of plasma leupeptin using the reversible binding of leupeptin to bovine pancreatic trypsin. | A competitive binding radioassay for leupeptin has been developed utilizing the reversible binding of leupeptin to bovine pancreatic trypsin. An ethanol precipitation step was introduced to separate trypsin-bound leupeptin from its free form. Advantages of this method are simplicity of the procedure and avoidance of the preparation of antiserum. The possible metabolites of leupeptin exhibit no significant inhibitory effect on leupeptin-trypsin binding in this system. This method was applied to the determination of plasma leupeptin levels in dogs after oral administration of the peptide. | Assay of plasma leupeptin using the reversible binding of leupeptin to bovine pancreatic trypsin. A competitive binding radioassay for leupeptin has been developed utilizing the reversible binding of leupeptin to bovine pancreatic trypsin. An ethanol precipitation step was introduced to separate trypsin-bound leupeptin from its free form. Advantages of this method are simplicity of the procedure and avoidance of the preparation of antiserum. The possible metabolites of leupeptin exhibit no significant inhibitory effect on leupeptin-trypsin binding in this system. This method was applied to the determination of plasma leupeptin levels in dogs after oral administration of the peptide. | 3,766,949 |
pubmed23n0125_13663 | Recovery of biologically functional messenger RNA from agarose gels by passive elution. | A general method for isolating biologically active messenger RNA (mRNA) from agarose gels is reported. Purified cellular RNA is resolved by preparative agarose gel electrophoresis and recovered in high yields (80%) by passive diffusion. Polyadenylated mRNA isolated from the eluted RNA is functionally intact based on the ability of the RNA to serve as a template in cell-free translation systems and complementary DNA synthesis reactions. The entire procedure is simple and rapid. A substantial purification of the mRNAs coding for skeletal muscle myosin heavy chain, light chain subunits and carbonic anhydrase III has been achieved employing this method. | Recovery of biologically functional messenger RNA from agarose gels by passive elution. A general method for isolating biologically active messenger RNA (mRNA) from agarose gels is reported. Purified cellular RNA is resolved by preparative agarose gel electrophoresis and recovered in high yields (80%) by passive diffusion. Polyadenylated mRNA isolated from the eluted RNA is functionally intact based on the ability of the RNA to serve as a template in cell-free translation systems and complementary DNA synthesis reactions. The entire procedure is simple and rapid. A substantial purification of the mRNAs coding for skeletal muscle myosin heavy chain, light chain subunits and carbonic anhydrase III has been achieved employing this method. | 3,766,950 |
pubmed23n0125_13664 | Tissue porphyrin pattern determination by high-speed high-performance liquid chromatography. | A rapid and sensitive method for the determination of porphyrin carboxylic acids in liver, kidney, and spleen by high-speed high-performance liquid chromatography is described. Porphyrins were extracted with recoveries greater than or equal to 98%, concentrated on disposable octadecylsilyl cartridges, and analyzed with a liquid chromatograph equipped with a 3 microns X 3 cm octadecylsilyl column and a fluorescence detector. Separation of di-, tetra-, penta-, hexa-, hepta-, and octacarboxylic acids was achieved within 5 min. The detection limits for uro, copro, and protoporphyrin were 20, 10, and 20 fmol, respectively. | Tissue porphyrin pattern determination by high-speed high-performance liquid chromatography. A rapid and sensitive method for the determination of porphyrin carboxylic acids in liver, kidney, and spleen by high-speed high-performance liquid chromatography is described. Porphyrins were extracted with recoveries greater than or equal to 98%, concentrated on disposable octadecylsilyl cartridges, and analyzed with a liquid chromatograph equipped with a 3 microns X 3 cm octadecylsilyl column and a fluorescence detector. Separation of di-, tetra-, penta-, hexa-, hepta-, and octacarboxylic acids was achieved within 5 min. The detection limits for uro, copro, and protoporphyrin were 20, 10, and 20 fmol, respectively. | 3,766,951 |
pubmed23n0125_13665 | Optimization of erythrocyte membrane glycoprotein fluorescent labeling with dansylhydrazine after polyacrylamide gel electrophoresis. | An improved procedure for the labeling of glycoproteins with dansylhydrazine subsequent to electrophoresis in polyacrylamide gels is reported. This procedure is derived from the work of Eckhardt et al. (1976, Anal. Biochem. 73, 192-197) and Weber and Hof (1975, Biochem. Biophys. Res. Commun. 65, 1298-1302) who showed that dansylhydrazine may be condensed with the aldehyde groups of oxidized glycoprotein carbohydrates and the resulting hydrazones reduced with dimethylamine borane and/or sodium borohydride. Using the known distribution of erythrocyte membrane glycoproteins as a benchmark the effect of variation of a number of process parameters was investigated and an optimal procedure identified. The procedure is shown to be relatively insensitive to moderate variations in reagent composition, pH, and time of incubation with dansylhydrazine solution or reducing agents. It is also shown that labeling patterns may be preserved in dried gels if dimethylsulfoxide is replaced or omitted from all of the process solutions and destaining is effected with 1 M sodium acetate, pH 5.6. While specifically developed for the labeling of erythrocyte membrane proteins, the procedure is demonstrated to be applicable to other glycoprotein containing preparations. | Optimization of erythrocyte membrane glycoprotein fluorescent labeling with dansylhydrazine after polyacrylamide gel electrophoresis. An improved procedure for the labeling of glycoproteins with dansylhydrazine subsequent to electrophoresis in polyacrylamide gels is reported. This procedure is derived from the work of Eckhardt et al. (1976, Anal. Biochem. 73, 192-197) and Weber and Hof (1975, Biochem. Biophys. Res. Commun. 65, 1298-1302) who showed that dansylhydrazine may be condensed with the aldehyde groups of oxidized glycoprotein carbohydrates and the resulting hydrazones reduced with dimethylamine borane and/or sodium borohydride. Using the known distribution of erythrocyte membrane glycoproteins as a benchmark the effect of variation of a number of process parameters was investigated and an optimal procedure identified. The procedure is shown to be relatively insensitive to moderate variations in reagent composition, pH, and time of incubation with dansylhydrazine solution or reducing agents. It is also shown that labeling patterns may be preserved in dried gels if dimethylsulfoxide is replaced or omitted from all of the process solutions and destaining is effected with 1 M sodium acetate, pH 5.6. While specifically developed for the labeling of erythrocyte membrane proteins, the procedure is demonstrated to be applicable to other glycoprotein containing preparations. | 3,766,952 |
pubmed23n0125_13666 | Simultaneous determination of tocopherols, ubiquinols, and ubiquinones in blood, plasma, tissue homogenates, and subcellular fractions. | A fast single-step lipid extraction procedure and high-performance liquid chromatography with in-line uv and electrochemical detection are used for the simultaneous quantitative determination of tocopherols, ubiquinols, and ubiquinones in blood, plasma, tissue homogenates, and subcellular fractions. The compounds of interest can be quantitatively extracted into hexane from a sodium dodecyl sulfate-treated aqueous homogenate after precipitation of protein by addition of an equal volume of ethanol. alpha-, gamma-, and delta-Tocopherol, ubiquinol 9, ubiquinol 10, and ubiquinones 9 and 10 can be well separated on a reversed phase column. Ubiquinones are detected at 275 nm by the uv detector, and ubiquinols and tocopherols by the electrochemical detector in the oxidative mode. Quantitation is done by comparing chromatographic peak heights to those of a standard solution containing known amounts of tocopherols, ubiquinols 9 and 10, and ubiquinones 9 and 10, analyzed under identical conditions. The high sensitivity of the electrochemical detection allows operation at low potentials (+0.5 V) with low detector response, but high selectivity for the easily oxidizable tocopherols and ubiquinols and decreased baseline noise. The uv detection limits the overall sensitivity of the procedure to 2 pmol ubiquinone, corresponding to 0.1 microM ubiquinone in the lipid extract. The ranges of values obtained for rat and guinea pig tissues, for rat liver mitochondria, and for blood and plasma from rats and humans are given. | Simultaneous determination of tocopherols, ubiquinols, and ubiquinones in blood, plasma, tissue homogenates, and subcellular fractions. A fast single-step lipid extraction procedure and high-performance liquid chromatography with in-line uv and electrochemical detection are used for the simultaneous quantitative determination of tocopherols, ubiquinols, and ubiquinones in blood, plasma, tissue homogenates, and subcellular fractions. The compounds of interest can be quantitatively extracted into hexane from a sodium dodecyl sulfate-treated aqueous homogenate after precipitation of protein by addition of an equal volume of ethanol. alpha-, gamma-, and delta-Tocopherol, ubiquinol 9, ubiquinol 10, and ubiquinones 9 and 10 can be well separated on a reversed phase column. Ubiquinones are detected at 275 nm by the uv detector, and ubiquinols and tocopherols by the electrochemical detector in the oxidative mode. Quantitation is done by comparing chromatographic peak heights to those of a standard solution containing known amounts of tocopherols, ubiquinols 9 and 10, and ubiquinones 9 and 10, analyzed under identical conditions. The high sensitivity of the electrochemical detection allows operation at low potentials (+0.5 V) with low detector response, but high selectivity for the easily oxidizable tocopherols and ubiquinols and decreased baseline noise. The uv detection limits the overall sensitivity of the procedure to 2 pmol ubiquinone, corresponding to 0.1 microM ubiquinone in the lipid extract. The ranges of values obtained for rat and guinea pig tissues, for rat liver mitochondria, and for blood and plasma from rats and humans are given. | 3,766,953 |
pubmed23n0125_13667 | Detection of metallothionein on nitrocellulose membrane using Western blotting technique and its application to identification of cadmium-binding proteins. | A convenient method for detecting metallothionein (MT) and other cadmium-binding proteins (Cd-BPs) in tissue cytosol was established. A sample that contains MT was separated on a sodium dodecyl sulfate-polyacrylamide gel and then electrophoretically transferred to a nitrocellulose membrane after reduction with 2-mercaptoethanol (2-ME) on the gel. The membrane was incubated in a buffer containing 109Cd and then subjected to autoradiography. MT-I and -II in rat liver cytosol were detected as radioactive bands together with a Cd-BP of 60,000 Da. On the other hand, three Cd-BPs of 40,000, 29,000, and 24,000 Da were detected in the cytosol when the reduction with 2-ME was omitted. | Detection of metallothionein on nitrocellulose membrane using Western blotting technique and its application to identification of cadmium-binding proteins. A convenient method for detecting metallothionein (MT) and other cadmium-binding proteins (Cd-BPs) in tissue cytosol was established. A sample that contains MT was separated on a sodium dodecyl sulfate-polyacrylamide gel and then electrophoretically transferred to a nitrocellulose membrane after reduction with 2-mercaptoethanol (2-ME) on the gel. The membrane was incubated in a buffer containing 109Cd and then subjected to autoradiography. MT-I and -II in rat liver cytosol were detected as radioactive bands together with a Cd-BP of 60,000 Da. On the other hand, three Cd-BPs of 40,000, 29,000, and 24,000 Da were detected in the cytosol when the reduction with 2-ME was omitted. | 3,766,954 |
pubmed23n0125_13668 | Rapid and effective transfer of integral membrane proteins from isoelectric focusing gels to nitrocellulose membranes. | A method describing the rapid and effective transfer of integral membrane protein from isoelectric focusing gels to nitrocellulose is described. Initial experiments were carried out with detergent-solubilized extracts of human erythrocyte membrane proteins. The effectiveness of the transfer was demonstrated by assaying for erythrocyte glucose transporter, an integral membrane protein, using specific antibodies followed by 125I-protein A and autoradiography. Several detergents including octyl glucoside, Triton X-100 and CHAPS were used in this study but only octyl glucoside effectively solubilized the glucose transporter and did not interfere with the electrotransfer of the protein. The glucose transporter separated on isoelectric focusing gels was effectively transferred after 2 h of electroblotting and was found to have an apparent pI of 6.4-6.5. These findings were substantiated by photolabeling red cell membranes with [3H]cytochalasin B in the presence or absence of D-glucose (which inhibits [3H]cytochalasin B binding to the glucose transporter) and separating the labeled proteins by two dimensional electrophoresis. With this procedure we identified a D-glucose sensitive 50-60 kDa protein focusing with an apparent pI of around pH 6.4-6.5. | Rapid and effective transfer of integral membrane proteins from isoelectric focusing gels to nitrocellulose membranes. A method describing the rapid and effective transfer of integral membrane protein from isoelectric focusing gels to nitrocellulose is described. Initial experiments were carried out with detergent-solubilized extracts of human erythrocyte membrane proteins. The effectiveness of the transfer was demonstrated by assaying for erythrocyte glucose transporter, an integral membrane protein, using specific antibodies followed by 125I-protein A and autoradiography. Several detergents including octyl glucoside, Triton X-100 and CHAPS were used in this study but only octyl glucoside effectively solubilized the glucose transporter and did not interfere with the electrotransfer of the protein. The glucose transporter separated on isoelectric focusing gels was effectively transferred after 2 h of electroblotting and was found to have an apparent pI of 6.4-6.5. These findings were substantiated by photolabeling red cell membranes with [3H]cytochalasin B in the presence or absence of D-glucose (which inhibits [3H]cytochalasin B binding to the glucose transporter) and separating the labeled proteins by two dimensional electrophoresis. With this procedure we identified a D-glucose sensitive 50-60 kDa protein focusing with an apparent pI of around pH 6.4-6.5. | 3,766,955 |
pubmed23n0125_13669 | High-performance liquid chromatographic separation of platinum complexes containing the cis-1,2-diaminocyclohexane carrier ligand. | Platinum drugs with the 1,2-diaminocyclohexane (dach) carrier ligand have shown great promise in cancer chemotherapy, but little is known about their metabolism in the body. Since it is possible to radiolabel the dach ligand, it should be possible to quantitate the biotransformation products of these drugs, provided a method were available to separate the biotransformation products. In this paper we describe a two-column high-performance liquid chromatography system which can be used to separate many likely dach-platinum biotransformation products from the parent compounds, and allow their identification. An initial separation on a reverse-phase Partisil ODS-3 column allowed resolution of the uncharged species. The peak fractions from this column were concentrated 10-fold and reinjected onto a cation exchange Partisil 10 SCX column to allow resolution of the positively charged species. This system allowed resolution of two prototype dach-platinum drugs, (cis-1,2-diaminocyclohexane)dichloroplatinum(II) and (cis-1,2-diaminocyclohexane)malonatoplatinum(II), the aquated species likely to form from these drugs, and the complexes formed when these compounds react with glutathione, metallothionein, and amino acids. By using cation exchange chromatography at pH 2.3 as well as pH 4 and by using 14C-labeled amino acids to determine stoichiometry, it was also possible to determine the most likely structures for some of the amino acid complexes. Most importantly, this system allowed clear separation of many of the likely biotransformation products tested from the biologically important aquated species. This system should prove useful for separating and identifying the biotransformation products of dach-platinum drugs in blood and urine, in tissue culture media, and inside the cell. | High-performance liquid chromatographic separation of platinum complexes containing the cis-1,2-diaminocyclohexane carrier ligand. Platinum drugs with the 1,2-diaminocyclohexane (dach) carrier ligand have shown great promise in cancer chemotherapy, but little is known about their metabolism in the body. Since it is possible to radiolabel the dach ligand, it should be possible to quantitate the biotransformation products of these drugs, provided a method were available to separate the biotransformation products. In this paper we describe a two-column high-performance liquid chromatography system which can be used to separate many likely dach-platinum biotransformation products from the parent compounds, and allow their identification. An initial separation on a reverse-phase Partisil ODS-3 column allowed resolution of the uncharged species. The peak fractions from this column were concentrated 10-fold and reinjected onto a cation exchange Partisil 10 SCX column to allow resolution of the positively charged species. This system allowed resolution of two prototype dach-platinum drugs, (cis-1,2-diaminocyclohexane)dichloroplatinum(II) and (cis-1,2-diaminocyclohexane)malonatoplatinum(II), the aquated species likely to form from these drugs, and the complexes formed when these compounds react with glutathione, metallothionein, and amino acids. By using cation exchange chromatography at pH 2.3 as well as pH 4 and by using 14C-labeled amino acids to determine stoichiometry, it was also possible to determine the most likely structures for some of the amino acid complexes. Most importantly, this system allowed clear separation of many of the likely biotransformation products tested from the biologically important aquated species. This system should prove useful for separating and identifying the biotransformation products of dach-platinum drugs in blood and urine, in tissue culture media, and inside the cell. | 3,766,956 |
pubmed23n0125_13670 | Effect of photoaffinity labeling on rabbit uterine progesterone receptor. | Photoaffinity labeling with [17 alpha-methyl-3H]promegestone ([ 3H]R5020) is an effective technique for the covalent labeling of the progesterone receptor (PR), which allows monitoring of the steroid receptor complex under denaturing conditions. The present study was initiated to evaluate whether photolabeled PR could be used also as a marker for PR under nondenaturing conditions. Accordingly, the effect of irradiation on each component of the reaction was evaluated separately. When [3H]R5020 alone was irradiated, there was a rapid (less than 5 min), light dependent destruction of [3H]R5020, as evident from increased formation of a more polar tritiated product on TLC and a concomitant decrease in the ability of the irradiated preparation to bind to PR. When rabbit uterine PR was irradiated in the absence of steroid, a gradual decrease in the binding capacity was observed, reaching 70% of the nonirradiated control in 10 min. The optimal irradiation time for covalent [3H]R5020-PR complex formation was determined by irradiation for up to 5 min, and separation of the products by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis. Specific labeling of proteins of Mr 116,000 and 85,000 was observed, with the rate of labeling of the two being similar, and reaching a plateau by 4 min of irradiation. The photolabeling efficiency ranged from 2 to 12%. Sucrose gradient ultracentrifugation of photolabeled PR revealed that both the irradiated sample and the nonirradiated control sedimented to the same position. Subsequent SDS-polyacrylamide gel electrophoresis of the sucrose gradient peak from the photolabeled sample showed the presence of both labeled proteins of Mr 116,000 and 85,000. In addition, photolabeled rabbit uterine PR (Mr 116,000 and 85,000) could be immunoprecipitated with a guinea pig antiserum raised against rabbit uterine PR. Analysis of the photoaffinity labeling procedure in our system revealed that the photodestruction of [3H]R5020 was very rapid. However, maximal labeling with [3H]R5020 was obtainable with minimal photodestruction of PR which suggests that photolabeled receptor can be used as a marker for PR under nondenaturing conditions. | Effect of photoaffinity labeling on rabbit uterine progesterone receptor. Photoaffinity labeling with [17 alpha-methyl-3H]promegestone ([ 3H]R5020) is an effective technique for the covalent labeling of the progesterone receptor (PR), which allows monitoring of the steroid receptor complex under denaturing conditions. The present study was initiated to evaluate whether photolabeled PR could be used also as a marker for PR under nondenaturing conditions. Accordingly, the effect of irradiation on each component of the reaction was evaluated separately. When [3H]R5020 alone was irradiated, there was a rapid (less than 5 min), light dependent destruction of [3H]R5020, as evident from increased formation of a more polar tritiated product on TLC and a concomitant decrease in the ability of the irradiated preparation to bind to PR. When rabbit uterine PR was irradiated in the absence of steroid, a gradual decrease in the binding capacity was observed, reaching 70% of the nonirradiated control in 10 min. The optimal irradiation time for covalent [3H]R5020-PR complex formation was determined by irradiation for up to 5 min, and separation of the products by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis. Specific labeling of proteins of Mr 116,000 and 85,000 was observed, with the rate of labeling of the two being similar, and reaching a plateau by 4 min of irradiation. The photolabeling efficiency ranged from 2 to 12%. Sucrose gradient ultracentrifugation of photolabeled PR revealed that both the irradiated sample and the nonirradiated control sedimented to the same position. Subsequent SDS-polyacrylamide gel electrophoresis of the sucrose gradient peak from the photolabeled sample showed the presence of both labeled proteins of Mr 116,000 and 85,000. In addition, photolabeled rabbit uterine PR (Mr 116,000 and 85,000) could be immunoprecipitated with a guinea pig antiserum raised against rabbit uterine PR. Analysis of the photoaffinity labeling procedure in our system revealed that the photodestruction of [3H]R5020 was very rapid. However, maximal labeling with [3H]R5020 was obtainable with minimal photodestruction of PR which suggests that photolabeled receptor can be used as a marker for PR under nondenaturing conditions. | 3,766,957 |
pubmed23n0125_13671 | A solid-phase substrate of heparanase: its application to assay of human melanoma for heparan sulfate degradative activity. | We found a tumor metastasis-associated heparan sulfate (HS)-degrading endoglycosidase in melanoma cells that is a unique endo-beta-glucuronidase (heparanase) capable of specifically cleaving HS at intrachain sites (M. Nakajima, T. Irimura, N. DiFerrante, and G. L. Nicolson, 1984, J. Biol. Chem. 259, 2283-2290). To perform rapid and microscale quantitative assays of heparanase we developed a solid-phase HS substrate by crosslinking radiolabeled HS onto agarose gel beads using one covalent linkage. The HS from bovine lung was partially N-desulfated and labeled with [14C]acetic anhydride. Free HS amino groups were completely acetylated, and reducing terminal saccharides were reductively aminated. The HS derivatives with amino groups at their reducing termini were coupled to amino-reactive agarose beads. Incubation of the solid-phase HS substrates with B16 melanoma cell extracts in the presence of D-saccharic acid 1,4-lactone (a potent exo-beta-glucuronidase inhibitor) resulted in the time- and dose-dependent release of [14C]HS fragments. Human melanoma cell lines were tested for HS-degrading endoglycosidase using the newly developed solid-phase HS substrates. The human malignant melanoma cells tested had high levels of HS-degrading activity that were comparable to those of highly metastatic murine B16-F10 melanoma cells. | A solid-phase substrate of heparanase: its application to assay of human melanoma for heparan sulfate degradative activity. We found a tumor metastasis-associated heparan sulfate (HS)-degrading endoglycosidase in melanoma cells that is a unique endo-beta-glucuronidase (heparanase) capable of specifically cleaving HS at intrachain sites (M. Nakajima, T. Irimura, N. DiFerrante, and G. L. Nicolson, 1984, J. Biol. Chem. 259, 2283-2290). To perform rapid and microscale quantitative assays of heparanase we developed a solid-phase HS substrate by crosslinking radiolabeled HS onto agarose gel beads using one covalent linkage. The HS from bovine lung was partially N-desulfated and labeled with [14C]acetic anhydride. Free HS amino groups were completely acetylated, and reducing terminal saccharides were reductively aminated. The HS derivatives with amino groups at their reducing termini were coupled to amino-reactive agarose beads. Incubation of the solid-phase HS substrates with B16 melanoma cell extracts in the presence of D-saccharic acid 1,4-lactone (a potent exo-beta-glucuronidase inhibitor) resulted in the time- and dose-dependent release of [14C]HS fragments. Human melanoma cell lines were tested for HS-degrading endoglycosidase using the newly developed solid-phase HS substrates. The human malignant melanoma cells tested had high levels of HS-degrading activity that were comparable to those of highly metastatic murine B16-F10 melanoma cells. | 3,766,958 |
pubmed23n0125_13672 | Separation of dimethylphosphatidates of alkylacyl glycerophosphocholine and their molecular species by high-performance liquid chromatography. | A high-performance liquid chromatography (HPLC) method for separation of the alkylacyl and diacyl analogs of choline glycerophospholipids (CGP) of guinea pig polymorphonuclear leukocytes and their molecular species is described. CGP were hydrolyzed with phospholipase D and then methylated with diazomethane to convert them to dimethylphosphatidates. The dimethylphosphatidates were then separated into the alkylacyl and diacyl subclasses by HPLC on a silica gel column within 15 min. The alkylacyl and diacyl analogs were then separated into individual molecular species by reverse-phase HPLC. Dimethylphosphatidates were resolved into 15 separate peaks, and 11-16 different molecular species of alkylacyl and diacyl glycerophosphocholines were identified on gas-liquid chromatography. The present results indicate that the CGP of polymorphonuclear leukocytes are composed of 27 major molecular species. In the alkylacyl subclass, the most predominant species was the 16:0-18:2 species (32%), followed by the 18:1-18:2 (18%), 16:0-16:0 (16%), and 16:0-18:1 (15%) species. The diacyl type consisted mainly of species with 18:2 at the 2-position, such as the 16:0-18:2, 18:0-18:2, and 18:1-18:2 species, the total percentage of which was 57%. | Separation of dimethylphosphatidates of alkylacyl glycerophosphocholine and their molecular species by high-performance liquid chromatography. A high-performance liquid chromatography (HPLC) method for separation of the alkylacyl and diacyl analogs of choline glycerophospholipids (CGP) of guinea pig polymorphonuclear leukocytes and their molecular species is described. CGP were hydrolyzed with phospholipase D and then methylated with diazomethane to convert them to dimethylphosphatidates. The dimethylphosphatidates were then separated into the alkylacyl and diacyl subclasses by HPLC on a silica gel column within 15 min. The alkylacyl and diacyl analogs were then separated into individual molecular species by reverse-phase HPLC. Dimethylphosphatidates were resolved into 15 separate peaks, and 11-16 different molecular species of alkylacyl and diacyl glycerophosphocholines were identified on gas-liquid chromatography. The present results indicate that the CGP of polymorphonuclear leukocytes are composed of 27 major molecular species. In the alkylacyl subclass, the most predominant species was the 16:0-18:2 species (32%), followed by the 18:1-18:2 (18%), 16:0-16:0 (16%), and 16:0-18:1 (15%) species. The diacyl type consisted mainly of species with 18:2 at the 2-position, such as the 16:0-18:2, 18:0-18:2, and 18:1-18:2 species, the total percentage of which was 57%. | 3,766,959 |
pubmed23n0125_13673 | Quantitation of picomole levels of N-acetyl- and N-glycolylneuraminic acids by a HPLC-adaptation of the thiobarbituric acid assay. | A simple HPLC adaptation of the periodate-TBA assay for free N-acetyl- and N-glycolylneuraminic acids greatly extends the sensitivity and increases the specificity of this standard colorimetric assay. The method, employing a C18 reverse-phase column eluted isocratically with a phosphoric acid-MeOH buffer, is linear over a range of 2 pmol to 20 nmol. Analyses can be performed directly on cell lysates and digests without prior purification of released sialic acids from contaminating salts and biological materials. Interference from 2-deoxysugars is completely eliminated as the chromophore from these compounds is completely resolved from that derived from sialic acids. The application of the technique to quantify cell-surface and total cellular TBA-reactive sialic acids on the surfaces of a variety of tumor cells is described. Additionally, the extent of desialylation of erythrocytes necessary to expose the T antigen is determined. | Quantitation of picomole levels of N-acetyl- and N-glycolylneuraminic acids by a HPLC-adaptation of the thiobarbituric acid assay. A simple HPLC adaptation of the periodate-TBA assay for free N-acetyl- and N-glycolylneuraminic acids greatly extends the sensitivity and increases the specificity of this standard colorimetric assay. The method, employing a C18 reverse-phase column eluted isocratically with a phosphoric acid-MeOH buffer, is linear over a range of 2 pmol to 20 nmol. Analyses can be performed directly on cell lysates and digests without prior purification of released sialic acids from contaminating salts and biological materials. Interference from 2-deoxysugars is completely eliminated as the chromophore from these compounds is completely resolved from that derived from sialic acids. The application of the technique to quantify cell-surface and total cellular TBA-reactive sialic acids on the surfaces of a variety of tumor cells is described. Additionally, the extent of desialylation of erythrocytes necessary to expose the T antigen is determined. | 3,766,960 |
pubmed23n0125_13674 | A high-performance liquid chromatographic method for the assay of cholesterol 7 alpha-hydroxylase activity. | A high-performance liquid chromatographic method has been developed for the measurement of cholesterol 7 alpha-hydroxylase activity in liver microsomes. 7 alpha-Hydroxycholesterol generated from endogenous cholesterol was derivatized with anthroyl 1-carbonitrile, chromatographed on a reverse-phase column, and detected fluorometrically. The detection limit of 7 alpha-hydroxycholesterol was 1 ng/tube. The cholesterol 7 alpha-hydroxylase activity in rat liver microsomes was assayed by this method, and the effects of some detergents and of the addition of exogenous cholesterol together with detergents on the enzyme activity were investigated. The endogenous 7 alpha- and 7 beta-hydroxycholesterol could be also measured by this method. | A high-performance liquid chromatographic method for the assay of cholesterol 7 alpha-hydroxylase activity. A high-performance liquid chromatographic method has been developed for the measurement of cholesterol 7 alpha-hydroxylase activity in liver microsomes. 7 alpha-Hydroxycholesterol generated from endogenous cholesterol was derivatized with anthroyl 1-carbonitrile, chromatographed on a reverse-phase column, and detected fluorometrically. The detection limit of 7 alpha-hydroxycholesterol was 1 ng/tube. The cholesterol 7 alpha-hydroxylase activity in rat liver microsomes was assayed by this method, and the effects of some detergents and of the addition of exogenous cholesterol together with detergents on the enzyme activity were investigated. The endogenous 7 alpha- and 7 beta-hydroxycholesterol could be also measured by this method. | 3,766,961 |
pubmed23n0125_13675 | A continuous-flow high-yield process for preparation of lipid-free hemoglobin. | Hypotonic hollow-fiber dialysis of bovine red blood cells followed by ultrafiltration through 0.1-micron pore hollow fibers provides a simple method for isolation of lipid-free hemoglobin. Hemoglobin (Hb) isolated by comparative techniques were all contaminated with membrane stroma. HPLC analysis of Hb revealed a protein peak of 99.6% purity and sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis revealed a single band. The process requires hypoosmotic dialysis of bovine RBC to a final 160-180 mosmol/kg osmotic pressure. Additional reduction in osmotic pressure causes irreversible cell lysis which leads to lipid contamination of the Hb. Processing of 1/2 liter of packed red blood cells requires 4-5 h, resulting in an average of 90% hemoglobin recovery. | A continuous-flow high-yield process for preparation of lipid-free hemoglobin. Hypotonic hollow-fiber dialysis of bovine red blood cells followed by ultrafiltration through 0.1-micron pore hollow fibers provides a simple method for isolation of lipid-free hemoglobin. Hemoglobin (Hb) isolated by comparative techniques were all contaminated with membrane stroma. HPLC analysis of Hb revealed a protein peak of 99.6% purity and sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis revealed a single band. The process requires hypoosmotic dialysis of bovine RBC to a final 160-180 mosmol/kg osmotic pressure. Additional reduction in osmotic pressure causes irreversible cell lysis which leads to lipid contamination of the Hb. Processing of 1/2 liter of packed red blood cells requires 4-5 h, resulting in an average of 90% hemoglobin recovery. | 3,766,962 |
pubmed23n0125_13676 | Determination of proteins and sulfobetaine with the Folin-phenol reagent. | This paper describes a method for the quantitative analysis of solutions containing a mixture of proteins and sulfobetaine. In a preliminary step the proteins, which interfere with the detergent assay, are separated by precipitation with trichloroacetic acid (8%). The insoluble fraction, dissolved in NaOH (1.0 N), and the soluble fraction, containing the detergent, are treated with the Folin-Ciocalteu phenol reagent, essentially following the method of O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall (1951, J. Biol. Chem. 193, 265-275). The absorbance of the protein fraction is read, as usual at 750 nm, while that of the detergent solution is read at 342 nm. At this wavelength, sulfobetaine, treated with the Folin reagent, absorbs strongly, the absorbances being proportional to its concentration up to 1.5 mg/ml. | Determination of proteins and sulfobetaine with the Folin-phenol reagent. This paper describes a method for the quantitative analysis of solutions containing a mixture of proteins and sulfobetaine. In a preliminary step the proteins, which interfere with the detergent assay, are separated by precipitation with trichloroacetic acid (8%). The insoluble fraction, dissolved in NaOH (1.0 N), and the soluble fraction, containing the detergent, are treated with the Folin-Ciocalteu phenol reagent, essentially following the method of O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall (1951, J. Biol. Chem. 193, 265-275). The absorbance of the protein fraction is read, as usual at 750 nm, while that of the detergent solution is read at 342 nm. At this wavelength, sulfobetaine, treated with the Folin reagent, absorbs strongly, the absorbances being proportional to its concentration up to 1.5 mg/ml. | 3,766,963 |
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