Development of a Simple and Cost-Effective Method Based on T7 Endonuclease Cleavage for Detection of Single Nucleotide Polymorphisms.
Single nucleotide polymorphisms (SNP) can be used as genetic markers and for risk assessment of allele-linked diseases, which can provide information for clinical diagnosis. Large-scale microarray and next-generation sequencing methods have made genome-wide SNP genotyping possible. However, in addition to their high cost, these techniques are dependent on having specialized equipment. Thus, there is a need for a simple genotyping method that can be implemented in a resource-limited environment. We developed a strategy for SNP genotyping based on T7 Endonuclease I cleavage and an enzyme-linked microparticle immune assay. Using this method, we genotyped two common SNP sites (rs11526468 and rs12979860). The quality of the genotyping process was validated. Although a 70% false-negative rate was observed, no false-positive reactions were found. Therefore, multiple parallel repeat reactions can offset the possibility of mutation detection failure. This method employs a duplicate reagent-dependent procedure, and therefore has the potential for integration into a portable kit for field utilization.


Oral mifepristone for chronic central serous chorioretinopathy.
Chronic central serous chorioretinopathy (CCSC) can result in permanent loss of vision. Unfortunately, many cases of CCSC are not eligible or do not respond to treatment with thermal laser or photodynamic therapy. Glucocorticoids have been implicated in the pathogenesis of central serous chorioretinopathy. Mifepristone, an oral glucocorticoid receptor antagonist, may be helpful in cases of CCSC. Mifepristone 200 mg was administered orally to 16 CCSC subjects in 2 separate protocols for up to 12 weeks. Visual acuity, examination, angiography, optical coherence tomography, and liver function were monitored during the treatment period. Favorable response to oral mifepristone was seen in CCSC patients with seven subjects gaining five or more letters of vision and seven subjects having improved optical coherence tomography findings. Treatment was well tolerated without serious adverse effects. Systemic glucocorticoid receptor antagonism with daily oral mifepristone does have a beneficial effect in treating some cases of CCSC. Further study is warranted.


Oxidative degradation of 2,6-dibromophenol using anion-exchange resin supported supramolecular catalysts of iron(III)-5,10,15,20-tetrakis (p-hydroxyphenyl)porphyrin bound to humic acid prepared via formaldehyde and urea-formaldehyde polycondensation.
An iron(III)-porphyrin catalyst, iron(III)-tetrakis(p-hydroxyphenyl)porphyrin (FeTHP), was introduced into a humic acid via a formaldehyde or urea-formaldehyde polycondensation reaction to stabilize the catalyst. The prepared supramolecular catalysts were then attached to Dowex-22, an anion-exchange resin. The oxidation of 2,6-dibromophenol (2,6-DBP) was then used, to evaluate the catalytic activities of the supported catalysts. The supported catalyst prepared via the urea-formaldehyde polycondensation reaction showed the highest catalytic activity of all catalysts tested. However, no debromination was observed under any conditions. To examine the reusability of the supported catalysts, they were evaluated on the basis of the decrease in the percent degradation of 2,6-DBP for the number times that they could be used. To determine why the catalytic activities decreased with increasing use, the surface of the supported catalysts were observed by scanning electron microscopy and energy dispersive X-ray spectrometry (SME-EDS) after each use. The poor reusability of the supported catalysts can be attributed to the fact that 2,6-DBP and/or brominated byproducts are tightly absorbed to the catalyst in the vicinity of the active site, which leads to inactivation of the supported catalysts.


A Qualitative Study of the Functional Outcomes Following First Metatarsophalangeal Joint (MTPJ) Arthrodesis Based on a Procedure Focused Questionnaire.
Hallux rigidus is the second most common condition to affect the first ray with an incidence of 2.5% in those above 50 years. Metatarsophalangeal Joint (MTPJ) arthrodesis remains the standard surgery. There are currently no patient-reported outcome measures or functional outcome measures specific to first MTPJ arthrodesis. Finding out what patients can and cannot do after surgery would help surgeons appropriately consent patients and manage expectations pre- and postsurgery. A pilot group of 15 patients postarthrodesis agreed on the suitability of the questions developed by the authors. As no further changes were made, a further 35 patients were recruited. Median age was 68 years, 78% were females, and 68% of patients were retired. Median follow-up was 64.5 months. Complete or almost complete pain relief was reported by 92% of patients. No major difficulty was reported by 97% of patients using ladders, 95% of patients driving, 90% of patients standing, 86% of patients wearing shoes without heels. Fifty-seven percent of patients reported extreme difficulty running and 48% of patients reported moderate or extreme difficulties wearing shoes with heels. None of the men reported difficulty with shoe wear without heels compared to 18% of women (p = .01). None of the men reported any difficulty in driving compared to 18% of women (p = .06). Difficulty in walking was reported in 44% of women compared to 9% of men (p = N/S). Our study is the first to reflect patients' own long term experiences following first MTPJ arthrodesis. Based on our study, following first MTPJ arthrodesis the majority of patients did not have trouble with pain, walking, standing, and driving. More than half of patients did not have trouble wearing shoes without heels; up to a third didn't have trouble wearing heels. More women experienced difficulty compared to men wearing shoes without heels, driving, and walking.


A qualitative analysis of family member needs and concerns in the population of patients with burns.
The importance of family support to ameliorate the recovery of a patient with burns has been documented in the literature. However, there is a dearth of research that identifies family members' needs and concerns during the hospitalization of patients with burns. Study aims were as follows: (1) identify support needs and concerns of family members of adult and pediatric patients currently in a burn center and (2) explore the relationship between family needs and the patient's severity of injury. An observational design was used that incorporated semistructured interviews with family members 1 to 3 days after the burn. A convenience sample of family members of burn center patients (n = 97) was recruited over 9 months. Content analysis was used to establish themes from interview data. Interrater reliability on coding of thematic units from 15% of the interviews was 86%; discrepancies were corrected to 100%. The average family member was 42.3 years old, female, white, and at least a high school graduate. Family members' concerns included general patient concerns, physical concerns about the patient, satisfaction of personal needs, and psychologic concerns about the patient. Major sources of support were family and friends, burn center staff, and spirituality. Family members of pediatric patients identified pain and skin graft surgery as priority worries. Satisfaction of personal needs, financial support, and information needs were greater among family members of intensive care unit patients in comparison with step-down patients. These findings underscore the importance of communication and reassurance between the burn team and the family members. Alterations to the support provided to families can be made on the basis of study subjects' responses.


Perinatal mortality in the United States: 1985-91.
In 1991 a total of 35,926 perinatal deaths occurred in the United States; this includes fetal deaths occurring at 28 weeks of gestation or later and infant deaths occurring under 7 days of age. The perinatal mortality rate was at a record low of 8.7 perinatal deaths per 1,000 live births and fetal deaths, 19 percent lower than the rate in 1985. Perinatal mortality rates differ by race. In 1991 the rates for the white and black populations were 7.4 and 15.7 perinatal deaths per 1,000 live births and fetal deaths, respectively. The rate for the Hispanic population was 7.9 compared with a rate of 7.1 for the non-Hispanic white population of an area comprised of 36 States and the District of Columbia. This area includes those States that reported Hispanic origin and whose reporting completeness was considered adequate for analysis.


Ozone tolerance in snap bean is associated with elevated ascorbic acid in the leaf apoplast.
Ascorbic acid (AA) in the leaf apoplast has the potential to limit ozone injury by participating in reactions that detoxify ozone and reactive oxygen intermediates and thus prevent plasma membrane damage. Genotypes of snap bean (Phaseolus vulgaris L) were compared in controlled environments and in open-top field chambers to assess the relationship between extracellular AA content and ozone tolerance. Vacuum infiltration methods were employed to separate leaf AA into extracellular and intracellular fractions. For plants grown in controlled environments at low ozone concentration (4 nmol mol-1 ozone), leaf apoplast AA was significantly higher in tolerant genotypes (300-400 nmol g-1 FW) compared with sensitive genotypes (approximately 50 nmol g-1 FW), evidence that ozone tolerance is associated with elevated extracellular AA. For the open top chamber study, plants were grown in pots under charcoal-filtered air (CF) conditions and then either maintained under CF conditions (29 nmol mol-1 ozone) or exposed to elevated ozone (67 nmol mol-1 ozone). Following an 8-day treatment period, leaf apoplast AA was in the range of 100-190 nmol g-1 FW for all genotypes, but no relationship was observed between apoplast AA content and ozone tolerance. The contrasting results in the two studies demonstrated a potential limitation in the interpretation of extracellular AA data. Apoplast AA levels presumably reflect the steady-state condition between supply from the cytoplasm and utilization within the cell wall. The capacity to detoxify ozone in the extracellular space may be underestimated under elevated ozone conditions where the dynamics of AA supply and utilization are not adequately represented by a steady-state measurement.


Prevalence and association of dental injuries with socioeconomic conditions and alcohol/drug use in adolescents between 15 and 19 years of age.
The aim of the present study was to investigate the prevalence of dental trauma, etiological factors, predisposing factors, and associations with socioeconomic status and the risk of alcohol and illicit drug use among adolescents in the city of Belo Horizonte, Brazil. A cross-sectional study was carried out that included clinical examinations and self-administered questionnaires. The sample population was composed of 891 adolescents from public and private schools. The Social Vulnerability Index (SVI) was used for socioeconomic classification. Information on alcohol and illicit drug use was obtained using two questionnaires: the Alcohol Use Disorders Identification Test (AUDIT) and the Alcohol, Smoking and Substance Involvement Screening Test (ASSIST). The prevalence of traumatic dental injury (TDI) was 24.7%. Falls (17.7%) was the most frequently cited etiological factor in dental injury. Among the participants with TDI, 32.8% were students in the private school system (P = 0.006). A total of 56.8% of individuals with accentuated overjet had some type of TDI (P = 0.000). There was a high prevalence of adolescents who consumed alcoholic beverages (50.3%) and used illicit substances (15.2%). However, no statistically significant associations were found between these variables and the presence of TDI. The results of the analysis demonstrate that individuals in a private school system [prevalence ratio (PR) = 1.11; 95% confidence interval (CI): 1.03-1.20] and those with accentuated overjet (>3 mm) (PR = 1.17; 95% CI: 1.10-1.25) had a 1.11- and 1.17-fold greater chance of belonging to the group of individuals diagnosed with some type of TDI. The prevalence of dental trauma in the study population was high. The same was true regarding alcohol and illicit drug use among the adolescents examined, although no statistically significant associations were found between these variables and a history of TDI. Private school system and accentuated overjet were significantly associated with dental trauma.


Quick and Sensitive UPLC-ESI-MS/MS Method for Simultaneous Estimation of Sofosbuvir and Its Metabolite in Human Plasma.
A simple, fast and highly sensitive RP-UPLC-MS/MS method was developed and validated for the simultaneous determination of sofosbuvir (SR) and its metabolite GS331007 in human plasma using ketotifen as an internal standard (IS). The separation was achieved on Acquity UPLC BEH C18 (50 × 2.1 mm, i.d. 1.7 µm, Waters, USA) column using acetonitrile:5 mM ammonium formate:0.1% formic acid (85:15:0.1% v/v/v) as a mobile phase at a flow rate of 0.35 mL/min in an isocratic elution. The Xevo TQD UPLC-MS/MS was operated under the multiple-reaction monitoring mode using positive electrospray ionization. Extraction with dichloromethane was used in the sample preparation. Method validation was performed as per the Food and Drug Administration (FDA) guidelines and the calibration curves of the proposed method were found to be linear in the range of 1-1000 ng/mL for SR and in the range of 10-1500 ng/mL for its metabolite (GS331007) with an elution time of 1.83 min. All validation parameters were within the acceptable range according to the bioanalytical methods validation guidelines. Furthermore, the obtained results of matrix effects indicate that ion suppression or enhancement from human plasma components was negligible under the optimized conditions. The proposed method can be applied in high-throughput analysis required for pharmacokinetic and bioequivalence studies in human samples.


Management of closed fractures.
A closed fracture is defined as a break in the continuity of the bone which does not communicate with the outside of the body. Management of the patient includes a thorough assessment of the fracture, evaluation of other injuries, and reduction as indicated. The patient is carefully observed for complications should problems arise.


Thyrotropin-releasing hormone stimulation tests in infants.
The TSH response to TRH administration (7 micrograms/kg) was measured in 68 infants (22 premature) who had abnormal thyroid screening tests by the filter paper method and whose serum thyroid function tests were only mildly abnormal. Twenty-eight infants (12 premature) had peak TSH values of 35 mU/L or less and were considered normal (group I). Forty infants (10 premature) had peak TSH values above 35 mU/L and were considered hyperresponsive (group II). The mean age at testing, screening T4, TSH levels that prompted the testing, as well as baseline T4, T3, and free T4 at the time of TRH testing were not different between the groups. The mean (+/- SD) baseline TSH value was greater in group II (6.8 +/- 2.3 mU/L) than in group I (4.4 +/- 2.2 mU/L; P < 0.001). However, there was a great deal of overlap in the individual TSH values (group I, 0.9-10 mU/L; group II, 1.9-10.6 mU/L). Mean peak TSH levels were significantly different in the two groups (group I, 24 +/- 7.7 mU/L; group II, 60.3 +/- 26.1 mU/L; P < 0.001). During long term follow-up, all 25 group I infants available for evaluation have been confirmed as clinically and biochemically normal. No infant diagnosed as normal was later found to have evidence of hypothyroidism. Fourteen infants in group II have had evidence of thyroid dysfunction. We conclude that the TSH response to TRH stimulation is a useful tool for the evaluation of infants suspected of having primary hypothyroidism. Whether hyperresponsiveness to TRH represents a form of neonatal hypothyroidism requiring treatment remains to be determined.


Disseminated Mycobacterium bovis Infection Complicated by Meningitis and Stroke: A Case Report.
We describe a case of a 19-year-old female presenting with Mycobacterium bovis meningitis, a rarely encountered infection. We discuss the use of pyrosequencing to aid in prompt diagnosis of M. bovis infection, as well as treatment strategies and challenges given the organism's intrinsic resistance to pyrazinamide.


Characterization of the two size forms of the alpha 1 subunit of skeletal muscle L-type calcium channels.
The molecular properties of two size forms of the alpha 1 subunit of purified skeletal muscle calcium channels were analyzed. The minor, full-length, form, alpha 1(212), was found to have an apparent molecular mass of 214 kDa by Ferguson plot analysis, while the major, truncated, form, now designated alpha 1(190), had an apparent molecular mass of 193 kDa. Antibody mapping of the C-terminal region of alpha 1(190) with 10 anti-peptide antibodies placed the C terminus between residues 1685 and 1699. Three consensus sites for cAMP-dependent protein phosphorylation are present in the C-terminal region of alpha 1(212) but not in alpha 1(190), and they may be important for the regulation of the ion conductance activity of the calcium channel.


Serum antibodies critically affect virus-specific CD4+/CD8+ T cell balance during respiratory syncytial virus infections.
Following infection with respiratory syncytial virus (RSV), reinfection in healthy individuals is common and presumably due to ineffective memory T cell responses. In peripheral blood of healthy adults, a higher CD4(+)/CD8(+) memory T cell ratio was observed compared with the ratio of virus-specific effector CD4(+)/CD8(+) T cells that we had found in earlier work during primary RSV infections. In mice, we show that an enhanced ratio of RSV-specific neutralizing to nonneutralizing Abs profoundly enhanced the CD4(+) T cell response during RSV infection. Moreover, FcγRs and complement factor C1q contributed to this Ab-mediated enhancement. Therefore, the increase in CD4(+) memory T cell response likely occurs through enhanced endosomal Ag processing dependent on FcγRs. The resulting shift in memory T cell response was likely amplified by suppressed T cell proliferation caused by RSV infection of APCs, a route important for Ag presentation via MHC class I molecules leading to CD8(+) T cell activation. Decreasing memory CD8(+) T cell numbers could explain the inadequate immunity during repeated RSV infections. Understanding this interplay of Ab-mediated CD4(+) memory T cell response enhancement and infection mediated CD8(+) memory T cell suppression is likely critical for development of effective RSV vaccines.


Spontaneous aortocaval fistula presenting with acute liver and renal failure: a case report.
Spontaneous aortocaval fistula is rare, occurring only in 3%-6% of all ruptured abdominal aortic aneurysms. A definitive diagnosis of aortocaval fistula is sometimes difficult, as the classic diagnostic signs (pulsatile abdominal mass with bruit, high-output heart failure and acute dyspnea) are present only in 20%-50% of all such cases. Pre-operative diagnosis is crucial, as adequate preparation has to be made for the massive bleeding expected at operation. Surgical repair of aortocaval fistula is now standardized repair of the fistula. We report herein a case of spontaneous aortocaval fistula, which presented with liver and renal failure.


Modular Evolution and Population Variability of Oikopleura dioica Metallothioneins.
Chordate Oikopleura dioica probably is the fastest evolving metazoan reported so far, and thereby, a suitable system in which to explore the limits of evolutionary processes. For this reason, and in order to gain new insights on the evolution of protein modularity, we have investigated the organization, function and evolution of multi-modular metallothionein (MT) proteins in O. dioica. MTs are a heterogeneous group of modular proteins defined by their cysteine (C)-rich domains, which confer the capacity of coordinating different transition metal ions. O. dioica has two MTs, a bi-modular OdiMT1 consisting of two domains (t-12C and 12C), and a multi-modular OdiMT2 with six t-12C/12C repeats. By means of mass spectrometry and spectroscopy of metal-protein complexes, we have shown that the 12C domain is able to autonomously bind four divalent metal ions, although the t-12C/12C pair -as it is found in OdiMT1- is the optimized unit for divalent metal binding. We have also shown a direct relationship between the number of the t-12C/12C repeats and the metal-binding capacity of the MTs, which means a stepwise mode of functional and structural evolution for OdiMT2. Finally, after analyzing four different O. dioica populations worldwide distributed, we have detected several OdiMT2 variants with changes in their number of t-12C/12C domain repeats. This finding reveals that the number of repeats fluctuates between current O. dioica populations, which provides a new perspective on the evolution of domain repeat proteins.


Wafer-scale two-dimensional semiconductors from printed oxide skin of liquid metals.
A variety of deposition methods for two-dimensional crystals have been demonstrated; however, their wafer-scale deposition remains a challenge. Here we introduce a technique for depositing and patterning of wafer-scale two-dimensional metal chalcogenide compounds by transforming the native interfacial metal oxide layer of low melting point metal precursors (group III and IV) in liquid form. In an oxygen-containing atmosphere, these metals establish an atomically thin oxide layer in a self-limiting reaction. The layer increases the wettability of the liquid metal placed on oxygen-terminated substrates, leaving the thin oxide layer behind. In the case of liquid gallium, the oxide skin attaches exclusively to a substrate and is then sulfurized via a relatively low temperature process. By controlling the surface chemistry of the substrate, we produce large area two-dimensional semiconducting GaS of unit cell thickness (∼1.5 nm). The presented deposition and patterning method offers great commercial potential for wafer-scale processes.


A polymer library approach to suicide gene therapy for cancer.
Optimal gene therapy for cancer must (i) deliver DNA to tumor cells with high efficiency, (ii) induce minimal toxicity, and (iii) avoid gene expression in healthy tissues. To this end, we generated a library of >500 degradable, poly(beta-amino esters) for potential use as nonviral DNA vectors. Using high-throughput methods, we screened this library in vitro for transfection efficiency and cytotoxicity. We tested the best performing polymer, C32, in mice for toxicity and DNA delivery after intratumor and i.m. injection. C32 delivered DNA intratumorally approximately 4-fold better than one of the best commercially available reagents, jetPEI (polyethyleneimine), and 26-fold better than naked DNA. Conversely, the highest transfection levels after i.m. administration were achieved with naked DNA, followed by polyethyleneimine; transfection was rarely observed with C32. Additionally, polyethyleneimine induced significant local toxicity after i.m. injection, whereas C32 demonstrated no toxicity. Finally, we used C32 to deliver a DNA construct encoding the A chain of diphtheria toxin (DT-A) to xenografts derived from LNCaP human prostate cancer cells. This construct regulates toxin expression both at the transcriptional level by the use of a chimeric-modified enhancer/promoter sequence of the human prostate-specific antigen gene and by DNA recombination mediated by Flp recombinase. C32 delivery of the A chain of diphtheria toxin DNA to LNCaP xenografts suppressed tumor growth and even caused 40% of tumors to regress in size. Because C32 transfects tumors locally at high levels, transfects healthy muscle poorly, and displays no toxicity, it may provide a vehicle for the local treatment of cancer.


Lack of expression of interleukin 8 and RANTES in autoimmune bullous skin diseases.
In autoimmune bullous skin diseases, accumulation of neutrophils and/or eosinophils in the affected skin represents a characteristic feature. So far, however, the induction of this granulocyte infiltration has not been elucidated. Regarding their biological effects, the chemokines interleukin 8 (IL-8) and RANTES (regulated upon activation normal T lymphocyte expressed and secreted) could play a role in this granulocyte accumulation. Immunohistochemical examination of lesional skin from patients with bullous pemphigoid, pemphigus, dermatitis herpetiformis and linear IgA disease was performed using a set of different antibodies against IL-8 and RANTES. Additionally, blister fluids were screened for soluble RANTES peptide using an ELISA. No difference in chemokine expression was found in lesions of autoimmune bullous diseases compared to normal skin. In contrast to chronic inflammatory diseases like psoriasis and eczema, where keratinocyte IL-8 immunoreactivity was found to differ from normal skin, keratinocyte immunoreactivity is not altered in autoimmune bullous diseases.


Outbreak of enteric microsporidiosis of hatchery-bred juvenile groupers, Epinephelus spp., associated with a new intranuclear microporidian in China.
A new enteric microsporidian was found to be associated with the mass mortality of hatchery-bred juvenile groupers, Epinephelus spp., in China. The outbreak usually occurred during the rainy season between May and November when water temperature ranged from 26 to 30 °C and salinity from 28 to 34 ppt, although this microsporidian can be detected year round. External clinical signs included severe emaciation, white faeces syndrome, anorexia, sinking to the bottom of culture ponds and mass mortality (up to 90%). Upon necropsy, severe intestinal oedema and thin and transparent intestinal wall could be observed. The mature spores are tiny, measuring 1.3-1.5 (1.35 ± 0.13) × 1.6-2.4 (2.16 ± 0.31) μm and can be found in the cytoplasm and the nucleoplasm of most enteric epithelial cells of host. Epidemiological investigation showed that this species was distributed throughout most of the culture area of grouper fingerlings in Fujian, Guangdong, Hainan and Guangxi provinces in China, with maximum prevalence of 95%. Molecular analysis based on the partial small subunit rRNA sequence (1045 bp) placed this species within the Enterocytozoonidae, but sequence identities to other species were below 90%. The exact taxonomic position warrants study of the ultrastructural characteristics of the developmental stages.


[Diagnosis of cochlear nerve foramen stenosis and atresia of the cochlear implantation].
To summarize methods on diagnosis of congenital cochlear nerve canal (CNC) stenosis or atresia and to report results of post-operation such as auditory electrophysiological test and speech test. Based on temporal bone HRCT and internal acoustic canal MRI, 27 bilateral congenital CNC stenosis/bony atresia cases were distinguished from 3 700 CI cases. Unilateral cochlear implantations were conducted above 27 cases. Post-operation tests such as auditory nerve response telemetry, EABR test, hearing threshold in sound field and speech recognition test were applied in the cases above. Incidence of CNC stenosis/bony atresia was observed at 0.73% (27 in 3700 CI cases). Impedance values were in the normal range accounting for 27 cases. Intro operative auditory nerve response telemetry were conducted accounting for 21 cases while 6 cases which were implanted with Combi40 + were not available for this test. Atypical ART response wave was observed for 14 cases, while no response for 7 cases. EABR test was completed in 27 cases within post-operation during 3 months and atypical EABR was identified in all cases. Hearing threshold in sound field was confirmed at 75 dB which was the average value of 500 Hz,1000 Hz,and 2000 Hz in all 27 cases. Speech recognition test result reached to both 65% (21 cases, simple finals test) and fewer than 10% (21 cases, simple initials test). By temporal bone HRCT pre-operation, CNC stenosis/bony atresia can be diagnosed. According to both relatively poor auditory and speech test results, further research and discussion are requisite to identify CI indication among the cases above.


Early development of stereotyped and self-injurious behaviors: II. Age trends.
Thirty-nine infants who were served in early intervention programs and who engaged in stereotyped or self-injurious behaviors were studied weekly from the time they entered the school until they were 3 years old. The development of these behaviors increased and declined over age more slowly than in typical babies. A few children retained the behaviors for long periods, and some were still showing them when they left the program. The hypothesis that body-rocking promotes motor development was not supported. Indeed, body-rocking may be a consequence of motor development. Head-banging seemed to be related to tantrums, whereas eye-poking developed early and seemed to be self-stimulatory. Suggestions for early treatment are presented.


Synthesis and cytotoxicity of novel pyrido[1,2-e]purines on multidrug resistant human MCF7 cells.
The anticancer activity of 4-methylaminopyridol[1,2-e]purine 6a, 4-(piperidin-1-yl)pyrido[1,2-e]purine 7a and their 7-methyl derivatives 6b, 7b was investigated against the human MCF7 cancer cell line in vitro. The sensitive cell line showed a range of sensitivities to 6a, 6b, 7a, 7b (IC50: 1.6 to 7.2 x 10(-4) M) and sensitivity to doxorubicin (IC50: 7.5 x 10(-7) M). A resistant cell line with the multidrug resistant phenotype was sensitive to these derivatives (IC50: 1.8 to 6.7 x 10(-4) M), doxorubicin (IC50: 5 x 10(-5) M) and drug activity seems to be not affected by MDR resistance. Our data show that 6a, 6b, 7a and 7b appear to exert a low cytotoxicity on sensitive and MDR resistant MCF7 human cancer cell lines.


African Charter on Human and Peoples' Rights.
This Convention contains provisions relating to women's rights, pay equity, health, education, the family, the aged, freedom of movement, and asylum. Algeria ratified the Convention on 20 March 1987, Benin on 25 February 1986, Botswana on 22 July 1986, Burkina Faso on 21 September 1984, Cape Verde on 6 August 1987, Central African Republic on 27 July 1986, Chad on 11 November 1986, Comoros on 18 July 1986, Egypt on 3 April 1984, Equatorial Guinea on 18 August 1986, Gabon on 26 June 1986, Guinea-Bissau on 6 March 1986, Libya on 26 March 1987, Sao Tome and Principe on 28 July 1986, Sierra Leone on 27 January 1984, Somalia on 20 March 1986, Sudan on 11 March 1986, Tanzania on 9 March 1984, Uganda on 27 May 1986, Zaire 28 July 1987, Zambia on 2 February 1984, and Zimbabwe on 12 June 1986.


Negotiating identities and social representations through intergroup contact in a community solidarity initiative.
In Ireland, displaced people experience segregation, discrimination, and disempowering regulations within the Direct Provision system. Community solidarity initiatives (CSI) aim to address the segregation and discrimination displaced people face through collaborative contact with residents/nationals of Ireland. However, asymmetric power relations mean that residents/nationals and displaced people are likely to experience intergroup contact differently, which has implications for identity negotiation. We investigated how displaced people and residents/nationals negotiated their identities and oriented to social representations in talk about their experiences of a CSI in the West of Ireland. We interviewed 17 displaced people and residents/nationals and conducted a thematic analysis, informed by Social Identity Approach and Social Representations Theory. Then, we applied a discursive approach to understand how participants constructed social identities and social representations. Our analysis produced two main themes: 'Identity negotiation strategies in talk about intergroup contact' and 'Understanding and orienting to intergroup boundaries'. We found that residents/nationals and displaced people negotiated their identities to maintain positive identification in relation to negative social representations about their groups. Participants also oriented to shared group representations in their talk , which has implications for the development of political solidarity between residents/nationals and displaced people.


Diet, exercise, behavior: the promise and limits of lifestyle change.
The prevalent surge in childhood and adolescent obesity within the past 3 decades poses a significant challenge for many pediatric clinicians who are charged with treating this condition. Comprehensive, research-based pediatric obesity treatment programs are essential to addressing this problem and preventing the transition of obesity and its many comorbidities into adulthood. This paper will identify dietary, physical activity, and behavioral approaches to lifestyle change and describe how they are incorporated as part of multidisciplinary treatment interventions in youth. Specific tailoring of treatment programs to address age and varying degrees of overweight and obesity will also be presented along with recommendations for future research.


[Preoperative drainage of the biliary tract in patients with obstructive jaundice].
The authors present their experience with preoperative external-internal percutaneous transhepatic drainage of the biliary pathways (PTD) in 143 patients. From this group in the final stage 53 patients were operated. The authors discuss the contemporary position and importance of preoperative PTD and compare the advantages of external-internal PTD and external drainage of the biliary pathways. A rapidly performed and correctly indicated external-internal PTD reduces effectively within several days the patient's bilirubin level, the bile passes into the duodenum, dehydration of the organism does not occur and vitamin K absorption is not impaired. All this has an impact on the success of surgery. Moreover, the external-internal drain modified in a suitable way surgical tactics and ensured effective drainage of the biliary pathways during the early postoperative period. If radical surgery is impossible, it is feasible as a suitable alternative procedure of palliative surgery to insert by the percutaneous route an expansible metal stent or leave the external-internal PTD drain in situ for a prolonged period. The authors discuss the criteria for indication, analyze the main problems of this procedure and submit their results.


Clinical and epidemiological characteristics of young patients with Peyronie's disease: a retrospective study.
The average age of men affected by Peyronie's disease (PD) is approximately 50-55 years, but cases have been reported even in adolescence. Several studies have already investigated the presence of PD in young men, and these studies reported a PD prevalence that varies between 1.5% and 10.8%. Having noticed a greater number of young patients in our centers in recent years, we decided to carry out a retrospective study to evaluate the prevalence of PD in patients aged <40 years, as well as to investigate any possible difference in evolution based on the age of PD patients. We selected a sample of patients (n=271) with a similar time of onset of disease. We then stratified all 271 patients into two groups: group A (age <40 years [n=46]) and group B (age ≥40 years [n=225]). All 271 patients were evaluated for the following variables: penile plaque volume, degree of penile curvature, penile pain, and erectile function. Plaque volume was measured in cm(3) by dynamic penile color Doppler sonography after administration of intracavernosal alprostadil 10 mcg. The number of younger patients was 46, accounting for 16.9% of the whole sample. Our study showed more frequent appearance and greater progression of penile curvature in younger patients. The average angle of penile curvature and average score of penile pain intensity in the younger men were significantly higher than in patients aged ≥40 years (P=0.025 and P=0.0001, respectively). In the younger patients, not only was the pain more intense (visual analog scale [VAS] of 5.2 versus 3.8), but it was also more frequently present than in patients aged ≥40 years (78.2% versus 62.2%) (P=0.042). We may conclude that since PD in young patients has a more acute onset and a greater possibility of progression, it should be treated conservatively as soon as it is diagnosed.


Mass Spectrometric Assays Reveal Discrepancies in Inhibition Profiles for the SARS-CoV-2 Papain-Like Protease.
The two SARS-CoV-2 proteases, i. e. the main protease (Mpro ) and the papain-like protease (PLpro ), which hydrolyze the viral polypeptide chain giving functional non-structural proteins, are essential for viral replication and are medicinal chemistry targets. We report a high-throughput mass spectrometry (MS)-based assay which directly monitors PLpro catalysis in vitro. The assay was applied to investigate the effect of reported small-molecule PLpro inhibitors and selected Mpro inhibitors on PLpro catalysis. The results reveal that some, but not all, PLpro inhibitor potencies differ substantially from those obtained using fluorescence-based assays. Some substrate-competing Mpro inhibitors, notably PF-07321332 (nirmatrelvir) which is in clinical development, do not inhibit PLpro . Less selective Mpro inhibitors, e. g. auranofin, inhibit PLpro , highlighting the potential for dual PLpro /Mpro inhibition. MS-based PLpro assays, which are orthogonal to widely employed fluorescence-based assays, are of utility in validating inhibitor potencies, especially for inhibitors operating by non-covalent mechanisms.


Regulation of folate and one-carbon metabolism in mammalian cells. IV. Role of folylpoly-gamma-glutamate synthetase in methotrexate metabolism and cytotoxicity.
Chinese hamster ovary (CHO) cells expressing human and Escherichia coli folylpoly-gamma-glutamate synthetase (FPGS) activities were used as models to study factors regulating the cytotoxicity and metabolism of methotrexate (MTX). CHO cells expressing human FPGS metabolized MTX to polyglutamates characteristic of human cells. Cellular MTX accumulation and metabolism to polyglutamates were dependent on the level of FPGS activity and were unaffected by putative gamma-glutamyl hydrolase inhibitors. The sensitivity of cells continuously exposed to MTX was not influenced by FPGS activity. After short term exposure to MTX, cells expressing higher levels of FPGS were more sensitive to the drug. MTX was not transported into the mitochondria and MTX treatment had no effect on preexisting mitochondrial folates while cytosolic folates were converted to oxidized forms. Mitochondrial folate accumulation was significantly impaired by MTX treatment, suggesting that the mitochondrial folate transport system is specific for reduced folates.


Aryl hydrocarbon receptor-mediated induction of Stearoyl-CoA desaturase 1 alters hepatic fatty acid composition in TCDD-elicited steatosis.
2,3,7,8-tetrachlorodibenzo-ρ-dioxin (TCDD) induces hepatic dyslipidemia mediated by the aryl hydrocarbon receptor (AhR). Stearoyl-CoA desaturase 1 (Scd1) performs the rate-limiting step in monounsaturated fatty acid (MUFA) synthesis, desaturating 16:0 and 18:0 into 16:1n7 and 18:1n9, respectively. To further examine the role of Scd1 in TCDD-induced hepatotoxicity, comparative studies were performed in Scd1(+/+) and Scd1(-/-) mice treated with 30 μg/kg TCDD. TCDD induced Scd1 activity, protein, and messenger RNA (mRNA) levels approximately twofold. In Scd1(+/+) mice, hepatic effects were marked by increased vacuolization and inflammation and a 3.5-fold increase in serum alanine aminotransferase (ALT) levels. Hepatic triglycerides (TRGs) were induced 3.9-fold and lipid profiling by gas chromatography-mass spectroscopy measured a 1.9-fold increase in fatty acid (FA) levels, consistent with the induction of lipid transport genes. Induction of Scd1 altered FA composition by decreasing saturated fatty acid (SFA) molar ratios 8% and increasing MUFA molar ratios 9%. Furthermore, ChIP-chip analysis revealed AhR enrichment (up to 5.7-fold), and computational analysis identified 16 putative functional dioxin response elements (DREs) within Scd1 genomic loci. Band shift assays confirmed AhR binding with select DREs. In Scd1(-/-) mice, TCDD induced minimal hepatic vacuolization and inflammation, while serum ALT levels remained unchanged. Although Scd1 deficiency attenuated TCDD-induced TRG accumulation, overall FA levels remained unchanged compared with Scd1(+/+) mice. In Scd1(-/-) mice, TCDD induced SFA ratios 8%, reduced MUFA ratios 13%, and induced polyunsaturated fatty acid ratios 5% relative to treated Scd1(+/+) mice. Collectively, these results suggest that AhR regulation of Scd1 not only alters lipid composition but also contributes to the hepatotoxicity of TCDD.


Paraquat poisoning: findings on chest radiography and CT in 42 patients.
The purpose of this study is to describe the radiologic manifestations of paraquat-induced pulmonary damage, with special emphasis on the sequential changes seen with this condition. Paraquat is a herbicide that has toxic effects on the lungs, liver, and kidneys. Progressive respiratory failure is a frequent cause of death. We analyzed retrospectively 42 patients with a history of paraquat ingestion and abnormal findings on chest radiographs. Radiographic changes during the first week after ingestion included diffuse consolidation (26/39), pneumomediastinum with or without pneumothorax (15/39), and cardiomegaly with widening of the superior mediastinum (8/39). Small cystic and linear shadows began to appear at the end of the first week and were the preponderant parenchymal abnormality after 2-4 weeks. Focal honeycombing was the major parenchymal abnormality after 4 weeks. High-resolution CT of the lung 9 months after paraquat exposure revealed localized fibrosis containing small cysts. Our study shows that the pulmonary manifestations of paraquat poisoning begin with air-space consolidation, which then leads to end-stage lung.


Transport of alkamides from Echinacea species through Caco-2 monolayers.
To gain more insights into the human intestinal absorption of alkamides from Echinacea species, transport studies were performed with the human adenocarcinoma colonic cell line Caco-2 (ATCC) as a model to assess the epithelial transport of dodeca-2 E,4 E,8 Z,10 E/ Z-tetraenoic acid isobutylamides (1/ 2). 30 minutes after apical loading of 25 microg/ml 1/ 2, about 15 % of these alkamides were detectable on the basolateral side. Close monitoring of the transport during 6 hours revealed a nearly complete transport to the basolateral side after 4 hours and no significant metabolism was observable. Transport experiments performed at 4 degrees C showed only a slight decrease in transport, which is a strong hint that dodeca-2 E,4 E,8 Z,10 E/ Z-tetraenoic acid isobutylamides (1/ 2) cross biological membranes by passive diffusion. Nearly the same results were obtained after preincubation of the Caco-2 cells with lipopolysaccharides (LPS) or phorbol 12-myristate-13-acetate (PMA) to mimic an inflammatory status. These results support the assumption that the alkamides can be easily transported from the intestinum and hence may contribute to the in vivo effects of Echinacea preparations.


[Binding of gonadotropin-releasing hormone (Gn-RH) to pituitary membranes in male rats].
The binding of gonadotropin-releasing hormone (Gn-RH) to membranes of male rats pituitary cells was studied. It appeared that Gn-RH was bound to 3 classes of sites and that the concentration in high affinity binding sites was very low. Moreover there was negatively cooperative interaction among these high affinity binding sites leading to a rapid decrease in binding affinity and consequently unmeasurable displacement of labeled hormone with large amounts of radio-inert ligand. These facts are inconsistent with the possible estimation of Gn-RH by radioreceptor assay.


Conformational preferences of proline analogues with a fused benzene ring.
The intrinsic conformational preferences of indoline-2-carboxylic acid (Inc) and its α-methylated derivative (αMeInc) have been investigated using quantum mechanical calculations. Specifically, the behavior of their N-acetyl-N'-methylamide derivatives, Ac-L-Inc-NHMe and Ac-L-αMeInc-NHMe, has been explored at the B3LYP/6-31+G(d,p) level. Such amino acids are analogues of proline and (α-methyl)proline, respectively, bearing a benzene ring fused to the C(γ)-C(δ) bond of the five-membered pyrrolidine ring. The additional aromatic group has been shown to significantly restrict the conformational space available to these residues by reducing the flexibility of both the five-membered cycle and the peptide backbone. The fused benzene ring also plays a critical role in determining the cis-trans arrangement of the amide bond involving the pyrrolidine nitrogen, which is also modulated by the presence of the α-methyl group in the αMeInc derivative. Furthermore, the influence of the environment on the conformational propensities of these compounds has been evaluated by using both a self-consistent reaction field model and a recently developed interface in a hybrid QM/MM scheme, in which the solvent molecules are treated explicitly with classical mechanics while the solute is described by quantum mechanics at the density functional theory level.


Personal strengths and traumatic experiences among institutionalized children given up at birth (Les Enfants de Duplessis--Duplessis' children): II: Adaptation in late adulthood.
In a companion article, we retrospectively examined the childhood strengths and adverse experiences of a group of orphans given up at or near birth and raised in Quebec institutions. This article examines the relationship of their early experiences to functioning and symptoms in later adulthood. The same follow-up interview of 81 adults (41 women, 40 men) at a mean age of 59.2 years included assessments of their current symptoms and functioning. The mean adult Social and Occupational Functioning Score (57.8; 95% CI, 54.7-61.0) indicated moderate difficulty. Psychiatric symptoms were significantly higher than in a matched population survey sample from Quebec. Mean overall defensive functioning indicated a neurotic (inhibited) level. Total trauma and childhood strengths predicted adult outcomes, but childhood strengths moderated the effects of trauma. Institutionalization of children--if unavoidable--must build in effective safeguards against adverse experiences, especially among children with few strengths, and foster children's strengths to avoid impaired adult outcomes.


Treating depression and pain.
Although chronic pain and depression have a high co-occurrence, major depression is underrecognized and undertreated in patients with chronic pain conditions, partly because these patients frequently present with somatic complaints rather than mood symptoms. Depression and pain have overlapping neurobiologic mechanisms, and a comprehensive treatment plan for patients with depression and painful physical symptoms should include pharmacotherapy to target serotonergic and noradrenergic neurotransmitters, along with cognitive-behavioral therapy, exercise, and patient education.


Age-related decreases in IL-2 production by human T cells are associated with impaired activation of nuclear transcriptional factors AP-1 and NF-AT.
Although transcriptional factors AP-1 and nuclear factor of activated T cells (NF-AT) are important for the normal induction of IL-2, it is unknown if the age-related decline in IL-2 production by activated human T cells may be associated with aberrancies in transcriptional regulatory proteins. In the current studies, IL-2 production by T cells from elderly (mean 78 years) and young (mean 37 years) humans was measured in cultures stimulated with PHA, PHA plus PMA, crosslinked anti-CD3 mAB OKT3 plus PMA, or PMA plus ionomycin. Substantial decreases of IL-2 production were observed for cell cultures from 7 of 12 elderly individuals in response to the different stimuli, whereas the levels of IL-2 produced by stimulated T cells from other elderly individuals were equivalent to those observed for stimulated T cells of young subjects. Analyses of nuclear extracts by electrophoretic DNA mobility shift assays showed that decreased IL-2 production by stimulated T cells of elderly individuals was closely associated with impairments in the activation of both AP-1 and NF-AT. By contrast, T cells from elderly subjects with normal levels of IL-2 production exhibited normal activation of AP-1 and NF-AT. In addition, the results of competition experiments analyzing the normal components of NF-AT showed that the age-related reductions in stimulus-dependent NF-AT complexes corresponded to the slow migrating complexes that were composed of c-Fos/c-Jun AP-1. The resting and stimulated levels of NF kappa B were reduced in T cells from certain elderly individuals; however, alterations of NF kappa B did not correlate with changes in IL-2 expression. Thus, these results show that age-related impairments in the activation of AP-1 and NF-AT are closely associated with decreased expression of IL-2 and further suggest that aberrancies in the signaling pathways important for the induction of transcriptionally active c-Fos/c-Jun AP-1 may contribute to the impaired activation of NF-AT.


[A case of recurrent breast cancer with chronic renal failure treated on an outpatient basis].
We report a case of recurrent breast cancer with chronic renal failure in a 58-year-old female. She could be treated on an outpatient basis under good general condition for most of her remaining life by chemoradiotherapy in combination with hemodialysis. Modification of the chemoradiotherapy procedures and collaboration of medical staff including the doctor in charge of home medical care and her family was indispensable in keeping her hospital stay as short as possible.


Childhood passive smoking, race, and coronary artery disease risk: the MCV Twin Study. Medical College of Virginia.
Children with long-term exposure to passive cigarette smoke may be at elevated risk for the development of premature coronary artery disease (CAD). To examine how CAD risk factors, exposure to passive smoking, sex, and race are related in pubertal children and to determine if there is an identifiable childhood risk profile (i.e., does passive smoking interact with other coronary risk factors to increase the risk of developing premature CAD). Cohort analytic study. The Medical College of Virginia (MCV) Twin Study, Richmond, Va. Randomly selected twins from 408 11-year-old twin pairs recruited from nearby schools. Data collection occurred at 18-month intervals on family and health histories, smoking and alcohol consumption, blood pressure, anthropometrics, and biochemical assays. Data from cohorts of 11-year-olds studied through age 15 years were analyzed by repeated-measures analyses of variance using a mixed modeling approach. Models for high-density lipoprotein cholesterol (HDL-C) included race, sex, passive smoking status, weight, systolic and diastolic blood pressures, and all interactions. Passive smoke exposure was greater in white families than in black families. Levels of HDL-C and HDL2-C (HDL subfraction 2 cholesterol) were lower in white children than in black children (visit 1: HDL-C, mean +/- SD, 1.21+/-0.26 vs. 1.31+/-0.26 mmol/L [47.0+/-10.1 vs. 50.6+/-10.1 mg/dL], P< or =.01; HDL2, mean +/- SD, 0.31+/-0.18 vs. 0.41+/-0.19 mmol/L [12.3+/-7.0 vs. 15.9+/-7.4 mg/dL], P< or =.001). Children with a family history of cardiovascular disease had differences in HDL-C levels related to race that were worsened by exposure to cigarette smoke. In these children, HDL-C level was lower in those exposed to passive smoking (visit 1: 1.18+/-0.23 vs. 1.25+/-0.23 mmol/L [45.6+/-9 vs. 48.2+/-9 mg/dL] and visit 4: 0.98+/-0.10 vs. 1.19+/-0.18 mmol/L [37.8+/-4 vs. 46.0+/-7 mg/dL]; P<.001), with white children having lower HDL-C levels than black children (visit 1: 1.12+/-0.21 vs. 1.36+/-0.23 mmol/L [43.2+/-8 vs. 52.7+/-9 mg/ dL] and visit 4: 0.97+/-0.31 vs. 1.01+/-0.31 mmol/L [37.6+/-12 vs. 39.0+/-12 mg/dL]; P = .004). In white families, as weight increased, boys exposed to passive smoking showed the greatest decrease in HDL-C level (P<.01 for weight by sex and passive smoking interaction). Risk factors for CAD, such as blood pressure, interacted with HDL-C and these relationships varied by race and sex. Pubertal children with long-term passive cigarette smoke exposure have lower HDL-C levels. Racial differences in HDL-C levels are related to passive smoke exposure. In children with a family history of cardiovascular disease, interactions exist between passive smoking, HDL-C level, and blood pressure that differ by sex and race. White males exposed to passive smoking who have a family history of cardiovascular disease and higher weights and diastolic blood pressures may be at special risk for premature CAD.


Central executive involvement in children's spatial memory.
Previous research with adults found that spatial short-term and working memory tasks impose similar demands on executive resources. We administered spatial short-term and working memory tasks to 8- and 11-year-olds in three separate experiments. In Experiments 1 and 2 an executive suppression task (random number generation) was found to impair performances on a short-term memory task (Corsi blocks), a working memory task (letter rotation), and a spatial visualisation task (paper folding). In Experiment 3 an articulatory suppression task only impaired performance on the working memory task. These results suggest that short-term and working memory performances are dependent on executive resources. The degree to which the short-term memory task was dependent on executive resources was expected to be related to the amount of experience children have had with such tasks. Yet we found no significant age-related suppression effects. This was attributed to differences in employment of cognitive strategies by the older children.


Aluminium chloride promotes anchorage-independent growth in human mammary epithelial cells.
Aluminium salts used as antiperspirants have been incriminated as contributing to breast cancer incidence in Western societies. To date, very little or no epidemiological or experimental data confirm or infirm this hypothesis. We report here that in MCF-10A human mammary epithelial cells, a well-established normal human mammary epithelial cell model, long-term exposure to aluminium chloride (AlCl(3) ) concentrations of 10-300 µ m, i.e. up to 100 000-fold lower than those found in antiperspirants, and in the range of those recently measured in the human breast, results in loss of contact inhibition and anchorage-independent growth. These effects were preceded by an increase of DNA synthesis, DNA double strand breaks (DSBs), and senescence in proliferating cultures. AlCl(3) also induced DSBs and senescence in proliferating primary human mammary epithelial cells. In contrast, it had no similar effects on human keratinocytes or fibroblasts, and was not detectably mutagenic in bacteria. MCF-10A cells morphologically transformed by long-term exposure to AlCl(3) display strong upregulation of the p53/p21(Waf1) pathway, a key mediator of growth arrest and senescence. These results suggest that aluminium is not generically mutagenic, but similar to an activated oncogene, it induces proliferation stress, DSBs and senescence in normal mammary epithelial cells; and that long-term exposure to AlCl(3) generates and selects for cells able to bypass p53/p21(Waf1) -mediated cellular senescence. Our observations do not formally identify aluminium as a breast carcinogen, but challenge the safety ascribed to its widespread use in underarm cosmetics.


Cost-effectiveness of posaconazole versus fluconazole for prevention of invasive fungal infections in U.S. patients with graft-versus-host disease.
The results of a pharmacoeconomic study of the cost-effectiveness of posaconazole versus fluconazole in preventing invasive fungal infections (IFIs) in patients with graft-versus-host disease (GVHD) are reported. The results of a randomized clinical trial suggested that posaconazole may be as effective as fluconazole for the prevention of IFIs in recipients of allogeneic cell transplants who develop severe GVHD and that posaconazole may be superior to fluconazole in reducing IFI-related deaths. Using published data from that trial and data from secondary sources, an economic model was developed to estimate the costs, IFIs avoided, and life-years saved with posaconazole versus fluconazole therapy. The results of the modeled 112-day treatment simulation were extrapolated to a lifetime horizon. In the modeled simulation, posaconazole therapy was associated with a lower probability of IFI development (0.05 versus 0.09), increased discounted life-years (7.87 life-years versus 7.66 life-years), and higher discounted costs per patient ($8,860 versus $5,710 in 2006 U.S. dollars) relative to fluconazole therapy. The estimated incremental cost-effectiveness of posaconazole versus fluconazole for IFI prophylaxis was $85,300 per IFI avoided and $15,300 per life-year saved. A sensitivity analysis indicated a 90% probability that the use of posaconazole for this purpose would be cost-effective at a threshold of $50,000 per life-year saved. Posaconazole is in the range of currently accepted criteria for cost- effectiveness relative to fluconazole for the prevention of IFIs among patients with GVHD.


Effects of educational intervention among reproductive age group women on safe abortion.
Many reproductive aged women needlessly die due to unsafe abortion even when they seek help to terminate their unwanted pregnancy. These deaths could have been prevented had they been aware that safe abortion service was available to them. The study aimed at finding out the effectiveness of the education intervention in improving knowledge among reproductive age group women regarding the safe abortion. An experimental intervention was carried out on safe abortion education among the reproductive age group women. The impact of pre- and post- intervention was evaluated by using a set of structured questionnaire in local language. The obtained data was analyzed by using the Excel and Statistical Package for the Social Sciences, version 12.0 for windows and result was interpreted. The post intervention finding revealed a significantly higher (p=0.001) mean on knowledge among participants about safe abortion compared to pre-observational test. The mean difference between the pre-test and post-test was 64.1% (Pre-test 11.18±12.88 Post-test 75.28±9.56). The research hypothesis was accepted with p value paired t-test at <0.001 (0.05). The result supports that the educational intervention was effective in increasing safe abortion awareness among reproductive aged group women. The safe abortion educational intervention program was instrumental to improve reproductive age women's knowledge considerably about safe abortion service.


Infection-induced autoantibodies and pregnancy related pathology: an animal model.
In addition to being the main cause of mortality worldwide, bacterial and viral infections can be the cause of autoimmune and pregnancy disorders as well. The production of autoantibodies during infection can be explained by various mechanisms, including molecular mimicry, bystander cell activation and epitope spreading. Conversely, bacterial and viral infections during pregnancy are especially dangerous for the fetus. It is documented that infection-induced inflammatory processes mediated by Toll-like receptors (TLR) represent the main cause of preterm labour. We used two crucial bacterial components and TLR ligands, namely peptidoglycan and lipopolysaccharide, to stimulate BALB/c mice before immunisation with tetanus toxoid. Tetanus toxoid is an inactive form of the toxin produced by bacterium Clostridium tetani and shares structural similarity with plasma protein β2-glycoprotein I. Treatment with peptidoglycan and lipopolysaccharide in combination with tetanus toxoid induced the production of pathological autoantibodies, different fluctuations in natural autoantibodies and different types of reproductive pathology in treated animals, with peptidoglycan treatment being more deleterious. We propose that the production of pathological autoantibodies, TLR activation and changes in natural autoantibodies play crucial roles in infection-induced reproductive pathology in our animal model.


Evaluation of meningococcal meningitis vaccination strategies for the meningitis belt in Africa.
Although the meningococcal polysaccharide vaccine has contributed to the control of Group A meningitis in the "meningitis belt" of Africa, recurrent large outbreaks have led to questions regarding vaccination strategy. We evaluated current and hypothetical vaccination strategies for the region. A model was formulated to analyze the effectiveness and costs of vaccine campaigns in response to outbreaks based on 7 years of weekly incidence data from Burkina Faso. Additional models analyzed the potential impact and costs of either a 1- or 4-dose routine scheduled delivery of meningococcal polysaccharide vaccine based on data reported to the World Health Organization from 16 countries during 1948 through 1996. Vaccine efficacy, vaccination coverage and economic data from literature reviews provided model assumptions. For Burkina Faso neither 1- nor 4-dose vaccination schedules would prevent >30% of meningitis cases compared with the 42% prevented through an outbreak response program of vaccinating districts, which reach an incidence of 15 per 100000 persons for 2 weeks. For the entire meningitis belt, routine coverage with the 1- or 4-dose schedule meningococcal vaccine would require 4.9 and 19.6 million doses annually, respectively, for an annual net cost of $4.4 to $12.3 million and prevent an average 10300 to 12600 cases (23 to 28%), assuming a long term vaccine efficacy of 50%. In addition an initial "catch-up" campaign costing up to $72 million to vaccinate the population from 1 to 30 years of age would be required before achieving that level of effectiveness. Given the relatively poor routine vaccination coverage in this region, current strategies of vaccination campaigns that achieve higher coverage would generally be more effective and less costly than the modeled routine scheduled programs, assuming that campaigns can be rapidly implemented. Until a better vaccine is available, countries in this region would be more efficient in improving the response times to outbreaks, perhaps through improved surveillance, and in bolstering existing vaccination infrastructures rather than embarking on strategies of questionable effectiveness.


Picking battles wisely: plant behaviour under competition.
Plants are limited in their ability to choose their neighbours, but they are able to orchestrate a wide spectrum of rational competitive behaviours that increase their prospects to prevail under various ecological settings. Through the perception of neighbours, plants are able to anticipate probable competitive interactions and modify their competitive behaviours to maximize their long-term gains. Specifically, plants can minimize competitive encounters by avoiding their neighbours; maximize their competitive effects by aggressively confronting their neighbours; or tolerate the competitive effects of their neighbours. However, the adaptive values of these non-mutually exclusive options are expected to depend strongly on the plants' evolutionary background and to change dynamically according to their past development, and relative sizes and vigour. Additionally, the magnitude of competitive responsiveness is expected to be positively correlated with the reliability of the environmental information regarding the expected competitive interactions and the expected time left for further plastic modifications. Concurrent competition over external and internal resources and morphogenetic signals may enable some plants to increase their efficiency and external competitive performance by discriminately allocating limited resources to their more promising organs at the expense of failing or less successful organs.


Blast from the past.
A recent death from diphtheria was a rare event in the UK. Vaccination has drastically reduced prevalence of this highly contagious infection.


Effects of VM26, a specific inhibitor of type II DNA topoisomerase, on SV40 chromatin replication in vitro.
We have examined the influence of VM26 (teniposide), a specific inhibitor of mammalian type II DNA topoisomerase, on the replication of SV40 minichromosomes in vitro. The replication system we used consists of replicative intermediate SV40 chromatin as substrate which is converted to mature SV40 chromatin in the presence of ATP, deoxynucleotides and a protein extract from uninfected cells. The addition of 100 microM VM26 to this system reduces DNA synthesis to 70 to 80 percent of the control and leads to an accumulation of 'late replicative intermediates'. The VM26 induced block of replication was not released by the addition of large quantities of type I DNA topoisomerase. We conclude, that type II DNA topoisomerase is essential for the final replication steps leading from late Cairns structures of replicative intermediates to monomeric minichromosomes. It appears that type I DNA topoisomerase can function as a swivelase during most of the replicative elongation phase, but must later be replaced by type II DNA topoisomerase.


The birth and early growth of a comprehensive multidisciplinary diabetes center in a community hospital: the Bildirici Center for Diabetes Care and Research at Laniado Hospital.
Diabetes is a multisystem disorder. Diabetes care is a multidisciplinary process. The Bildirici Center for Diabetes Care and Research opened its doors to its first patients in August 2007, as the first center in Israel to offer comprehensive integrated multidisciplinary care for people with diabetes. In addition to direct patient care, the Center has organized courses, lectures and workshops for people with diabetes, as well as for professionals caring for people with diabetes and for the general community. The ability of individuals within different departments to develop and implement such a center may have been facilitated by the smallness of our institution and the familiarity they share as a result. We have shown how a community hospital, without ties to a major medical center or academic institution, can establish a multidisciplinary inpatient and outpatient diabetes center, the first of its kind in a country.


Characterization of patients with bacteremia by methicillin-resistant Staphylococcus aureus in a high-complexity military hospital
Introduction: In Colombia, there are about 500,000 members in the national armed forces who consult military health institutions. In this population, methicillin-resistant Staphylococcus aureus (MRSA) has become a high-incidence pathogen. Objective: The aim of this study was to characterize patients with MRSA bacteremia in the Hospital Militar Central between 2012 and 2015. Materials and methods: This was an observational descriptive study with a retrospective review of clinical histories of hospitalized patients older than 18 years of age with positive blood cultures for methicillin-resistant S. aureus. The identification of the patients was made using the Whonet system, version 5.6. Results: From cultures positive for S. aureus, 24.8% were methicillin-resistant strains, with a higher prevalence in active military personnel. A similar frequency was observed for community-acquired MRSA bacteremias and those acquired at the hospital, with the community phenotype being the most frequent in both groups. The main infectious focus related to the development of bacteremia was soft tissue, followed by pulmonary tissue. There were higher complication rates in nosocomial bacteremias; 34.9% of the patients had prolonged stays attributable to complications triggered by the bacteremia. Conclusions: Active military personnel was the most affected population by MRSA, with a similar frequency in community-acquired and nosocomial bacteremias. The main infectious focus was soft tissue. Taking into account these data, studies that establish the prevalence of skin infections by MRSA should be carried out.


Heterogeneity of defensive responses after exposure to trauma: blunted autonomic reactivity in response to startling sounds.
Research on threat responses, particularly among trauma-exposed individuals, has traditionally focused on increased autonomic arousal and reactivity. However, clinical features associated with trauma exposure, such as dissociation (e.g., shutting down or "spacing out") manifest as the opposite pattern: non-reactivity and blunted arousal. These clinical features suggest that the possibility of threat responses other than fight/flight, namely, immobilization may be undergirded by hyper- or hypo-arousal. The goal of this paper is to examine autonomic responses to a stressful stimulus (acoustic startle) using analytic approaches which have been previously used to examine defensive responses before: heart rate acceleration, heart rate deceleration, and skin conductance response. We examined these responses in relation to symptoms (Posttraumatic Stress Disorder, or PTSD, and dissociation) and trauma exposure (cumulative exposure, age of onset) in a sample of trauma-exposed college students. We found evidence of blunted reactivity, with decreased acceleration and skin conductance, but with increased deceleration, particularly among individuals who had significant symptoms and early exposure to multiple types of trauma. However, individuals with sub-clinical symptoms and more attenuated exposure had large heart rate acceleration and skin conductance responses during the task. Taken together, these findings suggest that moderate symptoms and trauma exposure are related to exaggerated autonomic responses, while extreme symptoms and trauma exposure are related to blunted autonomic responses. These findings further suggest heterogeneity of stress responses within individuals with PTSD and with trauma exposure.


Unusual features of pomoviral RNA movement.
Potato mop-top pomovirus (PMTV) is one of a few viruses that can move systemically in plants in the absence of the capsid protein (CP). Pomoviruses encode the triple gene block genetic module of movement proteins (TGB 1, 2, and 3) and recent research suggests that PMTV RNA is transported either as ribonucleoprotein (RNP) complexes containing TGB1 or encapsidated in virions containing TGB1. Furthermore, there are different requirements for local or systemic (long-distance) movement. Research suggests that nucleolar passage of TGB1 may be important for the long-distance movement of both RNP and virions. Moreover, and uniquely, the long-distance movement of the CP-encoding RNA requires expression of both major and minor CP subunits and is inhibited when only the major CP sub unit is expressed. This paper reviews pomovirus research and presents a current model for RNA movement.


[Pterional orbital decompression in traumatic orbital hematomas].
The results are presented for pterional orbital decompression in 12 patients with symptomatic traumatic retrobulbar hematoma after various traumatic mechanisms. Pre- and postoperative course, neuroradiological findings, additional brain or facial injuries as well as outcome of eye function are analyzed in detail. Mean time delay between trauma and decompression was 56 h (2.4 days), with a wide range from 2 h to 15 days. Preoperative exophthalmos and pupillary disturbances as well as restrictions of extraocular movements decreased in all patients after orbital decompression and removal of the retrobulbar hematoma if the bleeding was localized. Visual acuity remained normal or showed significant improvement in seven patients, four of whom experienced complete recovery. In three patients the eye remained amaurotic. No complications related to the operation were seen. The pterional orbital decompression described here represents an effective alternative approach for patients with sight-threatening retrobulbar hematoma, especially in cases where it is necessary to gain space for the orbit in addition to evacuating space-occupying blood or bone clots and treating neighbouring lesions. Immediate detection and adequate treatment of orbital hematomas is mandatory to achieve an acceptable outcome of eye function.


Dammarane-type Saponins from Panax japonicus and their neurite outgrowth activity in SK-N-SH cells.
Four new dammarane-type saponins (1-4), named yesanchinosides G-J, together with nine ginsenosides (5-13) were isolated from Ye-Sanchi, the underground part of Panax japonicus collected in the south of Yunnan Province, China. Their structures were elucidated on the basis of spectroscopic and chemical methods. Ginsenosides Rb(1) and Rb(3) and notoginsenosides R(4) (6) and Fa (7) showed significant neurite outgrowth enhancing activities in human neuroblastoma SK-N-SH cells.


Self-Treatment Practices for Perceived Symptoms of Malaria in Ethiopia.
One of the most significant public health issues faced in Ethiopia is malaria. The most influential problem of this public health issue is overcoming barriers of having proper access to professional care and treatment. This study aims to elucidate the self-interventions undertaken by individuals when they perceive symptoms of malaria in a family-member prior to, or instead of, seeking care at a healthcare facility. Our study found that the prevalence of self-medication for malaria in the Wirtu Yedi kebele is 37.3%. Almost all individuals eventually sought treatment for malaria at the healthcare facility. More than half did so in less than one day. When self-treatment was used, there was a wide array of self-medication practices used, including modern medications not prescribed by a healthcare professional, herbs, and non-pharmacological measures. The more commonly used medications were chloroquine and Coartem® (artemether and lumefantrine). Most reported obtaining these medications at a drug outlet store without a prescription and prior to seeking care at a health center. Of the various herbs reported that were used to self treat malaria the most commonly used were garlic, ginger, and harmaguse. The use of herbs was found to be less common than the use of modern medication. Of the non-pharmacological interventions, rituals were the most prevalent.


Assessing meaningful change in quality of life over time: a users' guide for clinicians.
The objective of this article is to help clinicians interpret trial-based quality of life (QOL) changes over time. We address a series of questions and provide guidelines that are fundamental to assessing and interpreting change. The issues addressed are as follows: (1) What are the characteristics of the population for whom changes in QOL are reported? (2) Is the QOL questionnaire reliable, valid, and responsive to change? (3) Are the timing and frequency of assessments adequate? (4) Is the study adequately powered? (5) How are multiple QOL outcomes addressed in analyses? (6) How are multiple time points handled? (7) Can alternative explanations account for the observed change or lack of observed change (eg, handling of missing data, survival differences, and changes in patient's QOL perspective over time)? and (8) How is statistical significance translated into meaningful change? These guidelines will support clinicians in reviewing the clinical trial literature, which in turn can help them use the data in the treatment decision process.


Binding of spermidine and putrescine to energized liver mitochondria.
The binding of spermidine and putrescine to mitochondrial membranes was studied by applying a thermodynamic model of ligand-receptor interactions developed both for equilibrium and far-from-equilibrium binding processes (V. Di Noto, L. Dalla Via, A. Toninello, and M. Vidali Macromol. Theory Simul. 5, 165-181, 1996). Results demonstrate the presence of two monocoordinated binding sites (S1 and S2) for spermidine and one monocoordinated binding site (S2) for putrescine, all exhibiting high capacity and low affinity. It is proposed that differences in the polyamines' flexibility and hydrophilicity perhaps contributes to the observed variations in their interactions with the two sites. A comparison of the binding parameters of these polyamines with those of spermine reveals differences in the specific function of the S1 and S2 sites, identified in studies of spermine binding (L. Dalla Via, V. Di Noto, D. Siliprandi, and A. Toninello Biochim. Biophys. Acta 1284, 247-252, 1996).


Complex sampling: implications for data analysis.
Investigators in dental public health often use strategies other than simple random sampling to identify potential subjects; however, their statistical analyses do not always take into account the complex sampling mechanism. Often it is not clear whether a given strategy requires adjustment for stratification and/or cluster sampling of observations. We propose that the need for such adjustment depends on the primary study objective. As a general rule, we recommend that if the study goal is to estimate the magnitude of either a population value of interest (e.g., prevalence), or an established exposure-outcome association, adjustment of variances to reflect complex sampling is essential because obtaining appropriate variance estimates is a priority. However, if the study goal is to establish the presence of an association, especially in a preliminary investigation of novel conditions or understudied populations, obtaining appropriate variance estimates may not be of primary importance; hence, adjustment of variances for complex sampling is not always required, but often is recommended. This paper describes several types of complex sampling designs, methods of adjusting for complex sampling strategies, examples illustrating the effect of adjustment, and alternative approaches for analysis of complex samples.


Serum of normal subjects contains IgG with antibody-like specificity for phospholipids.
Using counterimmunoelectrophoresis, sera from normal subjects display precipitin activity against liposomes made by phosphatidic acid, phosphatidyl ethanolamine, phosphatidyl serine and cardiolipin. No activity is detected by using liposomes made with phosphatidyl choline, sphingomyelin and phosphatidyl inositol. The same pattern of precipitation is obtained when pure IgG from normal subjects is tested against phospholipids. No precipitin activity is found by using pure IgM. The precipitin reaction is prevented by preincubating liposomes with Fab obtained from normal IgG.


Evidence for autoinhibition of stimulation-induced noradrenaline release from vasa deferentia of the guinea-pig and rat.
Both phenoxybenzamine and idazoxan increased the efflux of radioactivity elicited by a train of stimulation (4 pulses at 5 Hz) in vasa deferentia preincubated with [3H]-noradrenaline. Phenoxybenzamine increased the release of radioactivity from vasa stimulated with a single pulse, whereas idazoxan did not. The contractile response in both guinea-pig and rat vasa was biphasic: phenoxybenzamine enhanced the initial twitch component and reduced the second component in guinea-pig vasa stimulated with a single pulse or a train of pulses. Idazoxan enhanced both phases of the response of guinea-pig vasa stimulated with a train of pulses but did not affect the response to stimulation with a single pulse. The effect of phenoxybenzamine in increasing the efflux of radioactivity produced by a single pulse of stimulation was abolished by cocaine, indicating that the increase in efflux was due to blockade of noradrenaline uptake. Contractile responses of guinea-pig vasa stimulated with a single pulse in the presence of cocaine were unaltered by phenoxybenzamine, whereas with a train of stimulation the twitch component was enhanced and the second phase was reduced. The effects of phenoxybenzamine or idazoxan on the efflux of radioactivity from rat vasa portions were qualitatively the same as were observed in whole vasa. The contractile response of the prostatic portion consisted of a rapid twitch with a single pulse of stimulation, but was biphasic with a train of stimulation; the response of the epididymal portion was biphasic with either a single pulse or a train of pulses. These results suggest that there is no inhibitory feedback modulation of noradrenaline release with a single pulse of stimulation in guinea-pig and rat vasa deferentia whereas, with a train of stimulation, there is autoinhibition of noradrenaline release.


The Sensitivity of Diagnostic Criteria of Marais S, et al. in Confirmed Childhood Tuberculous Meningitis.
To establish the sensitivity of the diagnostic criteria published by Marais and co-workers in 2010 for childhood tuberculous meningitis (TBM), a retrospective study on children with confirmed TBM was conducted. Between January 2006 and December 2019, children consecutively diagnosed with TBM were recruited retrospectively at our center. TBM was defined in cases where any of the following criteria were met: the presence of acid-fast bacilli (AFB) in cerebrospinal fluid (CSF) microscopy, CSF nucleic acid amplification test (NAAT, +), or M.tuberculosis cultured from CSF. The demographic and clinical features of all enrolled patients were recorded including clinical characteristics, CSF findings, cerebral imaging features, and other evidence of TB. A total of 30 children with confirmed diagnosis of TBM over an 14-year period were recruited. The mean age of patients was 7.2 ± 5.1 years and 16 (53.3%) were male. The estimated mean diagnostic score was 12.7 ± 2.4. Twenty-three (76.7%; 95% CI: 59.1-88.2%) patients were classified as "probable TBM" according to the Marais criteria and 7 (23.3%; 95% CI: 11.8-40.9%) as "possible TBM." Further statistical analysis revealed significant differences in CSF scores between probable and possible TBM groups. Other variables reported at a relatively low frequency, such as symptoms and imaging features, made little contribution to TBM diagnosis according to the Marais criteria. Childhood TBM could be effectively identified by the criteria defined by Marais et al. However, further revision is required to ensure that the system is more sensitive and easier to perform in practice.


Prediabetes in a nursing facility patient with renal insufficiency.
Prediabetes encompasses a variety of abnormalities, including impaired fasting glucose, impaired glucose tolerance, and metabolic syndrome. Prediabetes also increases the risk of developing type 2 diabetes mellitus (T2DM) by 3- to 10-fold, but the complications associated with hyperglycemia begin early in the patient's progression from normal glucose levels to diabetes. Early identification and treatment of prediabetes has the potential to reduce or delay progression to overt diabetes, to preempt related cardiovascular and microvascular disease, and to significantly improve morbidity and mortality. This clinical review provides a vehicle to examine management of prediabetes in patients 65 years of age and older, including diagnostic criteria and recommendations for management. Live symposium presentation based on clinical practice and research, medical literature, and studies published between May 1999 and March 2010 on managing diabetes in older adults, government statistics, and medical society guidelines. Effective prediabetes management is important to reducing the risk of progression to T2DM. Recommended first-line therapy is lifestyle modification that may include exercise, nutritional therapy, and weight loss. Pharmacological therapies, when indicated, can aid in improving glucose, blood pressure, and lipid parameters in this patient subgroup.


Mountain Cycling Ultramarathon Effects on Inflammatory and Hemoglobin Responses.
This study aimed to analyze the cumulative physiological burden of repetitive, strenuous exercise held during mountain cycling ultramarathon on regulatory mechanisms of hemoglobin degradation. Fifty-five nonprofessional athletes (mean age, 44.8 ± 7.1 yr) participating in a 9-consecutive-day mountain cycling ultramarathon (TransPortugal) underwent anthropometric, hematological, and biochemical assessments before and immediately after the race. Participants were further stratified as completers (nine courses) or noncompleters and were divided according to the time they took to complete the race. The heme oxygenase-1 (HMOX1) functional genetic polymorphism and haptoglobin (HP) phenotypic variants were also analyzed. Total leukocytes, neutrophil count, and monocyte count increased, whereas decreases in erythrocyte counts and hemoglobin were found between pre- and postultramarathon. Circulating haptoglobin (Hp) was increased, whereas its soluble receptor (sCD163) decreased. Athletes who completed all nine courses presented with increased leukocyte, neutrophil, and erythrocyte counts, as well as hemoglobin, red cell distribution width, total bilirubin, and total cholesterol levels. High-sensitivity C-reactive protein and Hp decreased in comparison with noncompleters. HMOX1 and HP genetic polymorphisms were associated with biochemical profile, notably with Hp levels. Analysis of covariance showed a significant effect of HP phenotype in Hp circulating levels at the end of race and on the magnitude of variation from pre- to postrace. Present findings support a comodulatory influence of genetic- and exercise-associated factors on resulting inflammatory and hemoglobin catabolic marker Hp after highly demanding endurance exercise.


Reversible "closing" of an electrode interface functionalized with a polymer brush by an electrochemical signal.
The poly(4-vinyl pyridine) (P4VP)-brush-modified indium tin oxide (ITO) electrode was used to switch reversibly the interfacial activity by the electrochemical signal. The application of an external potential (-0.85 V vs Ag|AgCl|KCl, 3M) that electrochemically reduced O(2) resulted in the concomitant consumption of hydrogen ions at the electrode interface, thus yielding a higher pH value and triggering the restructuring of the P4VP brush on the electrode surface. The initial swollen state of the protonated P4VP brush (pH 4.4) was permeable to the anionic [Fe(CN)(6)](4-) redox species, but the electrochemically produced local pH of 9.1 resulted in the deprotonation of the polymer brush. The produced hydrophobic shrunken state of the polymer brush was impermeable to the anionic redox species, thus fully inhibiting its redox process at the electrode surface. The interface's return to the electrochemically active state was achieved by disconnecting the applied potential, followed by stirring the electrolyte solution or by slow diffusional exchange of the electrode-adjacent thin layer with the bulk solution. The developed approach allowed the electrochemically triggered inhibition ("closing") of the electrode interface. The application of this approach to different interfacial systems will allow the use of various switchable electrodes that are useful for biosensors and biofuel cells with externally controlled activity. Further use of this concept was suggested for electrochemically controlled chemical actuators (e.g. operating as electroswitchable drug releasers).


Adverse events during use of intranasal desmopressin acetate for haemophilia A and von Willebrand disease: a case report and review of 40 patients.
We report our experience with the incidence of adverse events during the use of Stimate brand intranasal desmopressin acetate (IN DDAVP) for patients with haemophilia A (HA) or von Willebrand disease (vWD) after noting two severe adverse events in one adult patient. All patients with documented vWD (type 1 or 2 A) or haemophilia A (mild, moderate or symptomatic carrier) from the Emory Comprehensive Hemophilia Center who had IN DDAVP challenge testing or were using Stimate for treatment of bleeding were evaluated for adverse events by patient report or nursing observation of clinical signs and symptoms. Forty patients were studied. Sixty-eight per cent (27/40) experienced clinical signs and/or symptoms. The majority of these symptoms were mild, however several patients reported moderate to severe side-effects and one adult patient required medical intervention for symptomatic hyponatraemia. In our experience, two-thirds of patients tested experienced adverse signs and/or symptoms with the use of Stimate; considerably higher than that reported from preliminary results in the literature. Young age did not correlate positively with adverse reactions. Severe adverse events requiring medical intervention were rare, however symptoms such as moderate to severe headache, nausea, vomiting and weakness may necessitate evaluation for hyponatraemia. This is the first report of symptomatic hyponatraemia in an adult patient with recommended dosing of Stimate. Side-effects may be minimized if patients adhere to instructions regarding fluid intake and composition while using IN DDAVP.


Multi-shot echo-planar diffusion tensor imaging in cervical spondylotic myelopathy.
The aim of this study was to use diffusion tensor imaging (DTI) to investigate changes in diffusion metrics in patients with cervical spondylotic myelopathy (CSM) up to five years after decompressive surgery. We correlated these changes with clinical outcomes as scored by the Modified Japanese Orthopedic Association (mJOA) method, Neck Disability Index (NDI), and Visual Analogue Scale (VAS). We used multi-shot, high-resolution, diffusion tensor imaging (ms-DTI) in patients with cervical spondylotic myelopathy (CSM) to investigate the change in diffusion metrics and clinical outcomes up to five years after anterior cervical interbody discectomy and fusion (ACDF). High signal intensity was identified on T2-weighted imaging, along with DTI metrics such as fractional anisotropy (FA). MJOA, NDI, and VAS scores were also collected and compared at each follow-up point. Spearman correlations identified correspondence between FA and clinical outcome scores. Significant differences in mJOA scores and FA values were found between preoperative and postoperative timepoints up to two years after surgery. FA at the level of maximum cord compression (MCL) preoperatively was significantly correlated with the preoperative mJOA score. FA postoperatively was also significantly correlated with the postoperative mJOA score. There was no statistical relationship between NDI and mJOA or VAS. ms-DTI can detect microstructural changes in affected cord segments and reflect functional improvement. Both FA values and mJOA scores showed maximum recovery two years after surgery. The DTI metrics are significantly associated with pre- and postoperative mJOA scores. DTI metrics are a more sensitive, timely, and quantifiable surrogate for evaluating patients with CSM and a potential quantifiable biomarker for spinal cord dysfunction. Cite this article: Bone Joint J 2020;102-B(9):1210-1218.


Analysis of farnesyl transferase activity during hormone-induced maturation of Xenopus laevis oocytes.
Preincubation of Xenopus laevis oocytes with insulin or insulin-like growth factor 1 (IGF-1) resulted in inhibition of farnesyl transferase (FTase) activity measured both in vivo (after microinjection of tritiated farnesyl pyrophosphate and Ras-CVIM into oocytes) and in extracts using a filtration assay. FTase activity measured in oocyte extracts was inhibited 55% after a 20 min treatment of oocytes with 1 microM insulin or 10 nM IGF-1. The apparent IC(50) for inhibition of oocyte FTase by IGF-1 is 0.3 nM. The observed decrease in FTase activity was apparently not due to translocation of enzyme from cytosol to membrane, since activities measured both in soluble extracts and resuspended crude pellets displayed comparable levels of inhibition following hormone treatment. Using a hexapeptide (TKCVIM) as substrate, FTase activity was also inhibited 65% when oocytes were pretreated with 10 nM IGF-1. Two FTase inhibitors [(alpha-hydroxyfarnesyl) phosphonic acid (HFPA) and chaetomellic acid A (CA)] effectively inhibited Xenopus oocyte FTase by 80-90% when added to assay mixtures (IC(50) values of 338 +/- 96 nM HFPA and 232 +/- 80 nM CA) or after incubation of oocytes in drug before preparation of soluble extracts for assay (IC(50) values of 7 +/- 6 nM HFPA and 328 +/- 128 nM CA). The farnesyl transferase inhibitors were observed to slow the time course of oocyte maturation but did not block the IGF-1-induced maturation response. J. Exp. Zool. 286:193-203, 2000.


No neocentric activity on Aegilops markgrafii chromosome E.
Classical neocentromeres, proven mainly in Poaceae species, cause meiotic drive in higher plants. Here I present data indicating that the morphological stretching of a chromatin domain at the long arm of Aegilops markgrafii chromosome E during meiosis is not due to a supposed neocentric activity. The stretching is visible at the alien univalent in metaphase I and anaphase I pollen mother cells of Triticum aestivum--Ae. markgrafii monosomic addition line E and at one univalent of the 49-chromosome-containing plants containing the haploid Ae. markgrafii genome added to wheat. The absence of both centromeric histone variant CENH3 and attachment of spindle fibers, proven by immunostaining with antibodies against CENH3 and tubulin, respectively, revealed that the univalent elongation of chromosome E is not caused by a classical plant neocentromere.


Evaluation of photodynamic activity of C60/2-hydroxypropyl-β-cyclodextrin nanoparticles.
The objective of this study is to evaluate the ability of C(60)/2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) naonparticles to generate reactive oxygen species (ROS) and to induce cell toxicity by the photoirradiation. C(60) nanoparticles were prepared by cogrinding with HP-β-CyD for 3 h at 4°C under reduced pressure. The photodynamic activity of C(60)/HP-β-CyD nanoparticles was evaluated by spectroscopic methods, including the electron spin resonance spin-trapping method, and by the cell viability test using Hela cells. C(60)/HP-β-CyD nanoparticles efficiently generated not only superoxide anion radical (O(2)(·-)) and hydroxyl radical (·OH), but also singlet oxygen ((1)O(2)) through photoirradiation. The ROS generation was enhanced by decreasing the mean particle diameter of C(60) nanoparticles, and the particle size smaller than 90 nm showed a high generation of ·OH and (1)O(2). In addition, HP-β-CyD enhanced the generation of (1)O(2), compared with polyvinylpyrrolidone (an effective solubillizer for C(60)), due to partial disposition of C(60) in the hydrophobic CyD cavity. Furthermore, C(60) /HP-β-CyD nanoparticles showed cell toxicity after the light irradiation, but no toxicity was observed without the light irradiation. Therefore, HP-β-CyD is useful for the preparation of stable C(60) nanoparticles with high ROS generation ability, and C(60)/HP-β-CyD nanoparticles are a promising photosensitizer for photodynamic therapy.


Prognostic value of Bcl-2 in invasive breast cancer receiving chemotherapy and endocrine therapy.
Apoptosis induced by anticancer agents is a mechanism of treatment activity, overexpression of antiapoptotic genes could produce drug resistant tumors. We have demonstrated that the susceptibility of Bcl-2-negative tumors to a series of anticancer drugs was significantly higher than that of Bcl-2-positive tumors. The purpose of this study was to examine if negative Bcl-2 expression has treatment benefit in breast cancer patients received chemotherapy and endocrine treatment. A cohort of 147 Japanese women with invasive ductal carcinoma was studied. All the patients received postoperative adjuvant therapy consisting of combination chemotherapy with cyclophosphamide, epirubicin, and fluorouracil, and tamoxifen therapy. The expression of Bcl-2, estrogen receptor (ER) and MIB-1 was evaluated in breast cancer surgical specimens. Bcl-2 immunostaining was inversely correlated with increasing histologic grade (p=0.0095) and MIB-1 (p=0.0128). Furthermore, a positive correlation between Bcl-2 and ER was observed (p<0.0001). Unexpectedly, survival curves determined by the Kaplan-Meier method and univariate analysis demonstrated that Bcl-2-positivity was associated with favorable prognosis (p=0.0430). Cox proportional hazard model demonstrated that positive Bcl-2 expression still has favorable survival (p=0.0242) after consideration of other prognostic factors. Our results imply that Bcl-2 is a significant favorable prognostic factor for breast cancer with chemotherapy and endocrine therapy.


Employer-sponsored health insurance and prescription drug coverage for new retirees: dramatic declines in five years.
Employer-sponsored health insurance is often described as the most reliable private source of Medicare supplementation, particularly for prescription drug benefits. This study's findings show that employer coverage is becoming an increasingly less dependable source of coverage for new retirees, and the problem is likely to get worse. We found that the proportion of Medicare beneficiaries ages 65-69 with employer coverage declined from 46 percent in 1996 to 39 percent in 2000. The proportion with drug coverage from an employer declined from 40 percent in 1996 to 35 percent in 2000. Losses among males, the group most affected, would have been even greater had it not been for a slight increase in benefits from spouses' policies.


The Use of NaOH Solutions for Fouling Control in a Membrane Bioreactor: A Feasibility Study.
Nowadays, the microbial production of 1,3-propanediol (1,3-PD) is recognized as preferable to the chemical synthesis. However, finding a technological approach allowing the production of 1,3-PD in the membrane bioreactor (MBR) is a great challenge. In the present study, a ceramic ultrafiltration (UF) membrane (8 kDa) for treatment of 1,3-PD broths was used. It has been demonstrated that the membrane used provides the stable permeate flux that is necessary to ensure the stability of the fermentation process in MBR technology. It was noticed that the broth pH has a significant impact on both the final 1,3-PD concentration and permeate flux. Moreover, the feasibility of using NaOH for fouling control in the MBR was evaluated. It has been shown that 1% NaOH solution is effective in restoring the initial membrane performance. To the best of our knowledge, this study is the first to shed light onto the possibility of reducing the amount of the alkaline solutions generated during the MBR operation. Indeed, it has been found that 1% NaOH solution can be successfully used several times for both membrane cleaning and to stabilize the broth pH. Finally, based on the results obtained, the technological conceptions of the MBR technology were designed.