[Expression of recombinant human doppel protein and analysis of its cytotoxic activities].
Doppel (Dpl) is a newly identified PrP-associated protein. In this study, specific primers for human PRND gene were designed based on the human PRND cDNA sequence encoding human Dpl protein reported in the GenBank. The full-length PRND gene sequence with 531 bp long was obtained from DNA of human peripheral leucocytes as the template by polymerase chain reaction (PCR). After verified by sequence analysis, the PCR product was inserted into a prokaryotic-expressing vector and then transformed into E. coli JM109. The recombinant human Doppel protein (rhDpl) was expressed as inclusion bodies after IPTG induction, with the yield of more than 60% of total bacterial proteins. The rhDpl protein was purified by Ni2+ affinity chromatography and cleaved by hydroxylamine. SDS-PAGE revealed the molecular weight of the purified rhDpl protein was about 15 kD. Trypan blue and MTT assays identified that rhDpl in vitro inhibited the growth of human neuroblastoma cell line SH-SY5Y at concentrations > or =50 microg/mL and human cervical cancer cell line HeLa at concentrations > or =100 microg/mL, showing remarkably dose-and time-dependant manners. Hoechst33342-staining of SH-SY5Y cells treated with rhDpl showed massive apoptosis under fluorescent microscope. These results indicate that Dpl protein possesses cytotoxic activity in vitro, with obvious tissue-specific characteristics. This study provides the foundation for further study of Dpl biological functions in vitro and in vivo.


Objectively measured physical activity in Finnish employees: a cross-sectional study.
To objectively measure the amount of intensity-specific physical activity by gender and age with respect to body mass index (BMI) during workdays and days off among Finnish employees. A cross-sectional study. Primary care occupational healthcare units. A sample of 9554 Finnish employees (4221 men and 5333 women; age range 18-65 years; BMI range 18.5-40 kg/m(2)) who participated in health assessments related to occupational health promotion. The amount of moderate-to-vigorous (MVPA) and vigorous (VPA) physical activity (≥3 and ≥6 metabolic equivalents, respectively) was assessed by estimating the minute-to-minute oxygen consumption from the recorded beat-to-beat R-R interval data. The estimation method used heart rate, respiration rate and on/off response information from R-R interval data calibrated by age, gender, height, weight and self-reported physical activity class. The proportion of participants fulfilling the aerobic physical activity recommendation of ≥150 min/week was calculated on the basis of ≥10 min bouts, by multiplying the VPA minutes by 2. Both MVPA and VPA were higher among men and during days off, and decreased with increasing age and BMI (p<0.001 for all). Similar results were observed when the probability of having a bout of MVPA or VPA lasting continuously for ≥10 min per measurement day was studied. The total amount of VPA was low among overweight (mean ≤2.6 min/day), obese (mean ≤0.6 min/day) and all women in the age group 51-65 years (mean ≤2.5 min/day) during both types of days. The proportion of participants fulfilling the aerobic physical activity recommendation was highest for normal weight men (65%; 95% CI 62% to 67%) and lowest for obese women (10%; 95% CI 8% to 12%). Objectively measured physical activity is higher among men and during days off, and decreases with increasing age and BMI. The amount of VPA is very low among obese, overweight and older women.


Expanding functional repertoires of fungal peroxisomes: contribution to growth and survival processes.
It has long been regarded that the primary function of fungal peroxisomes is limited to the β-oxidation of fatty acids, as mutants lacking peroxisomal function fail to grow in minimal medium containing fatty acids as the sole carbon source. However, studies in filamentous fungi have revealed that peroxisomes have diverse functional repertoires. This review describes the essential roles of peroxisomes in the growth and survival processes of filamentous fungi. One such survival mechanism involves the Woronin body, a Pezizomycotina-specific organelle that plugs the septal pore upon hyphal lysis to prevent excessive cytoplasmic loss. A number of reports have demonstrated that Woronin bodies are derived from peroxisomes. Specifically, the Woronin body protein Hex1 is targeted to peroxisomes by peroxisomal targeting sequence 1 (PTS1) and forms a self-assembled structure that buds from peroxisomes to form the Woronin body. Peroxisomal deficiency reduces the ability of filamentous fungi to prevent excessive cytoplasmic loss upon hyphal lysis, indicating that peroxisomes contribute to the survival of these multicellular organisms. Peroxisomes were also recently found to play a vital role in the biosynthesis of biotin, which is an essential cofactor for various carboxylation and decarboxylation reactions. In biotin-prototrophic fungi, peroxisome-deficient mutants exhibit growth defects when grown on glucose as a carbon source due to biotin auxotrophy. The biotin biosynthetic enzyme BioF (7-keto-8-aminopelargonic acid synthase) contains a PTS1 motif that is required for both peroxisomal targeting and biotin biosynthesis. In plants, the BioF protein contains a conserved PTS1 motif and is also localized in peroxisomes. These findings indicate that the involvement of peroxisomes in biotin biosynthesis is evolutionarily conserved between fungi and plants, and that peroxisomes play a key role in fungal growth.


Blood-retina barrier dysfunction in experimental autoimmune uveitis: the pathogenesis and therapeutic targets.
Experimental autoimmune uveitis (EAU), an animal model of human uveitis, is characterized by infiltration of autoimmune T cells in the uvea as well as in the retina of susceptible animals. EAU is induced by the immunization of uveitogenic antigens, including either retinal soluble-antigen or interphotoreceptor retinoid-binding proteins, in Lewis rats. The pathogenesis of EAU in rats involves the proliferation of autoimmune T cells in peripheral lymphoid tissues and breakdown of the blood-retinal barrier, primarily in the uvea and retina, finally inducing visual dysfunction. In this review, we describe recent EAU studies to facilitate the design of a therapeutic strategy through the interruption of uveitogenic factors during the course of EAU, which will be helpful for controlling human uveitis.


Degenerative joint disease.
We have focused on several features somewhat unique to OA which require special attention relative to clinical trials. These include disease classification, for which the schema in Table 1 was developed, patient selection and admission criteria, and the choice of appropriate clinical outcome parameters. We suggest the following: That future trials be limited to patients with idiopathic OA with specified joint groups involved. That pain, assessed with a visual analogue scale, and both the patient's and investigator's global assessment be the primary clinical outcome variables to be analysed. That concomitant analgesic agents not be used during anti-inflammatory drug trials to prevent confounding pain relief as an outcome variable. That future, long-term studies use validated health status measures to assess functional outcomes.


Tryptophan and the innate intestinal immunity: Crosstalk between metabolites, host innate immune cells, and microbiota.
The intestinal mucosal barrier is critical for the absorption of nutrients and the health of both humans and animals. Recent publications from clinical and experimental studies have shown the importance of the nutrients-bacteria-host interaction for the intestinal homeostasis. Dysfunction of these interactions has been reported to be associated with metabolic disorders and development of intestinal diseases such as the irritable bowel syndrome and inflammatory bowel diseases. Tryptophan and its metabolites, including kynurenine, kynurenic acid, and 5-hydroxytrptamine, can influence the proliferation of enterocytes, intestinal integrity, and immune response, as well as intestinal microbiota, therefore, regulating and contributing to the intestinal health. In this review, we highlight recent findings on the effect of tryptophan and its metabolites on the mucosal barrier and intestinal homeostasis and its regulation of innate immune response. Moreover, we present the signaling pathways related to Trp metabolism, such as mammalian target of rapamycin, aryl hydrocarbon receptor, and pregnane X receptor, which contribute to the intestinal homeostasis and discuss future perspectives on spontaneous interference in host tryptophan metabolism as potential clinical strategies of intestinal diseases.


Effect of mental health collaborative care models on primary care provider outcomes: an integrative review.
Collaborative care models (CCMs) have robust research evidence in improving mental health outcomes for diverse patient populations with complex health care needs. However, the impact of CCMs on primary care provider (PCP) outcomes are not well described. This integrative review synthesizes the evidence regarding the effect of mental health CCMs on PCP outcomes. PubMed, CINAHL, Web of Science, and PsycInfo were systematically searched using key terms, with inclusion criteria of English language, peer-reviewed literature, primary care setting, PCP outcomes, and mental health CCM. This resulted in 1,481 total records, with an additional 14 records identified by review of reference lists. After removal of duplicates, 1,319 articles were reviewed based on title and abstract, 190 full-text articles were assessed, and a final selection of 15 articles were critically appraised and synthesized. The articles included a wide variety of sample sizes, designs, settings, and patient populations, with most studies demonstrating low or moderate quality evidence. Although CCMs had an overwhelmingly positive overall effect on PCP outcomes such as knowledge, satisfaction, and self-efficacy, multiple logistical barriers were also identified that hindered CCM implementation such as time and workflow conflicts. Adaptability of the CCM as well as PCP enthusiasm enhanced positive outcomes. Newer-to-practice PCPs were more likely to participate in CCM initiatives. Accumulating evidence supports CCM expansion, to improve both patient and PCP outcomes. Logistical efforts may enhance CCM adaptability and workflow. Further studies are needed to specifically examine the effect of CCMs on PCP burnout and retention.


[Inhibitive effects of chitosan on proliferation of fibroblasts in vitro].
To investigate the effects of chitosan on the cell cycle of the human fibroblasts and on the Ki-67 antigen expression in vitro and to investigate the mechanism of chitosan preventing the postoperative tissue adhesion. The cultured fibroblasts were treated for 48 hours with 0, 0.01, 0.1, 1.0, 10.0 mg/ml of chitosan, respectively; then, the cell cycle of the fibroblasts was measured by the flow cytometry. The cultured fibroblasts were treated for 24 hours with the chitiosan at the above concentrations; then, the Ki-67 antigen in the cell nucleus was detected with the immunohistochemical staining to observe its expression. The growth of the fibroblasts was obviously suppressed by chitosan, especially in the cell morphology. When the concentrations of chitosan were 1.0 mg/ ml and 10.0 mg/ml, the percentages of the fibroblasts in the proliferation stage were 32.3%+/- 5.2% and 14.7% +/- 2.9%, respectively, which were significantly smaller than the percentage of the fibroblasts when the concentration of chitosan was 0 mg/ml (the control group) (41.9% +/-5.8%, P<005). When the concentrations were 0. 01 mg/ml and 0. 1 mg/ml, the percentages of the fibroblasts in the proliferation stage were 39.0%+/-6.0% and 35.5%+/- 3.4%, respectively, which were smaller than that of the control, but not significantly different from that of the control (P> 0.05). When the concentrations of chitosan were 0.1 mg/ml, 1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts that had the positive Ki-67 antigen were 37.3% +/- 3.4%, 30.5% +/- 6.2% and 17.8% +/- 3.0%, respectively, which were significantly smaller than that of the control (57.6% +/-8.9%, P<0.05). When the concentration was 0. 01 mg/ml, the percentage of the fibroblasts that had the positive Ki-67 antigen was 54.1% +/- 8.0%, which was smaller than that of the control, but not significantly different from that of the control (P>0.05). Chitosan can inhibit the proliferation of the fibroblasts and increase the percentage of the fibroblasts in the quiescent stage, which can be considered as one of the mechanisms that chitosan can prevent the postoperative tissue adhesion.


Nuclear magnetic resonance studies on the role of intrinsic metals in Escherichia coli RNA polymerase. Effect of DNA template on the nucleotide-enzyme interaction.
Nuclear magnetic resonance studies were performed to investigate the effect of DNA template on the interaction of initiating nucleotide ATP with Escherichia coli RNA polymerase (RPase) in which one of the two intrinsic Zn ions was substituted with a Co(II) (Co-Zn RPase) or Mn(II) (Mn-Zn RPase) ion. This intrinsic metal ion is located at the initiation site in the beta subunit of RPase. The paramagnetic effects of Co-Zn and Mn-Zn RPases on the relaxation rates of 1H- and 31P-nuclei of ATP were used to determine the distances from the intrinsic metal to various atoms of ATP bound at the initiation sites in the presence of DNA. The distances from the metal to H2, H8, H1', alpha-P, beta-P, and gamma-P atoms were estimated to be 6.7 +/- 0.9, 4.1 +/- 0.6, 6.0 +/- 1.2, 7.5 +/- 0.8, 9.4 +/- 1.0, and 9.8 +/- 1.0 A, respectively. These distances were compared with those measured in the absence of DNA (Chatterji, D., and Wu, F. Y.-H. (1982) Biochemistry 21, 4657). In both the presence and absence of DNA, the close proximity between the intrinsic metal and the H8 atom strongly indicates that the metal is coordinated directly to the base moiety of ATP. Such a coordination may provide a structural basis for the selection of a purine nucleotide during the initiation process. The presence of DNA causes the H2 atom to move away (greater than 2 A) from the intrinsic metal, whereas all three phosphorus atoms shift closer (greater than 3 A) toward the metal. The possible mechanistic implications of the conformational alteration of ATP at the initiation site induced by the DNA template is discussed.


Clonal chromosomal defects in the molecular pathogenesis of refractory hyperparathyroidism of uremia.
Indirect X chromosome-inactivation analyses have demonstrated that most parathyroid glands from patients with uremic refractory secondary/tertiary hyperparathyroidism are monoclonal neoplasms. However, little is known regarding the specific acquired genetic abnormalities that must underlie such clonal expansion or the molecular pathogenetic features of this disorder, compared with primary parathyroid adenomas. To address these issues in a uniquely powerful manner, both comparative genomic hybridization (CGH) and genome-wide molecular allelotyping were performed with a large group of uremia-associated parathyroid tumors. As indicated by CGH, one or more chromosomal changes were present in 24% of the tumors, which is markedly different from the value for common sporadic adenomas (72%). Two recurrent abnormalities that had not been previously described for sporadic parathyroid adenomas were noted with CGH, i.e., gains on chromosomes 7 (9%) and 12 (11%). Losses on chromosome 11 occurred in only one of the 46 uremia-associated tumors (2%); the tumor also contained a somatic mutation of the remaining MEN1 allele (221del18). A total of 13% of tumors demonstrated recurrent allelic loss on 18q, with 18q21.1-q21.2 being defined as the putative tumor suppressor-containing region. In conclusion, the powerful combination of genome-wide molecular allelotyping and CGH has identified recurrent clonal DNA abnormalities that suggest the existence and locations of genes important in uremic hyperparathyroidism. In addition, genome-wide patterns of somatic DNA alterations, including disparate roles for MEN1 gene inactivation, indicate that markedly different molecular pathogenetic processes exist for clonal outgrowth in severe uremic hyperparathyroidism versus common parathyroid adenomas.


Isolated Ileal Perforation in Infancy: A Lethal Initial Presentation of Hirschsprung's Disease.
Arare case of ileal perforation, as a fatal initial presentation of total colonic aganglionosis (TCA) in infancy is reported. A 10-week-old boy, was brought to the emergency department with symptoms of complicated intestinal obstruction. He looked ill, was lethargic, markedly dehydrated and had a severely distended abdomen. An abdominal X-ray revealed multiple air fluid levels seen in a distended small intestine. During exploratory laparotomy the ileum was massively dilated with distal segment perforation. Ileal perforation repair was performed. A totally collapsed microcolon was identified. Biopsies were taken from the high rectum, sigmoid and hepatic flexure. Appendectomy and ileostomy were performed. All biopsies, as well as the appendix, showed absence of ganglion cells. Despite this procedure the patient progressively deteriorated and later died due to sepsis. Ileal perforation in infants is a rare, but potentially fatal initial presentation of TCA. Early detection is essential to prevent life-threatening complications.


EXPERIENCE WITH MANAGING RETROSTERNAL GOITRES IN IBADAN, NIGERIA.
There is no general consensus on the definition of retrosternal goitre (RSG) however thyroidectomy remains the gold standard of treatment with or without a sternotomy. To review the outcome of surgical management of retrosternal goitres. Retrospective review of records of patients who had thyroidectomy for RSG over a 15-year period. Out of a total of 45 patients, 34(76%) were females and 11(24%) were males with a male/female ratio of 3:1; while their age ranged between 28 and 72years with a mean of 57+15SD. All the patients were euthyroid and a quarter did not have symptoms apart from a neck mass. In all, 15% of the patients had recurrent goitre. CT scan of neck and chest was done in 31 (72%) patients; while 44 (98%) patients had cervical retrosternal goitres, 1(2%) patient had ectopic retrosternal goitre. A cervical incision was sufficient in 28 (62%) patients while 17 (38%) patients required additional sternotomy. Total thyroidectomy was done in all the patients. There were post-operative complications in 19 (42%) patients. Histopathology showed that 3(6.6%) patients had papillary thyroid carcinoma while 42(93.4%) had benign pathology findings. Surgical removal is the treatment of choice. Most retrosternal goitres can be resected through a collar stud incision; however the possibility of a need for a sternotomy should always be planned. The simultaneous occurrence of cervical and ectopic retrosternal goitre should always be ruled out with a CT scan.


Application of remifentanil combined with propofol in the diagnosis of colon cancer with awakening painless digestive endoscopy.
Anesthetic effect of remifentanil combined with propofol in awakening painless endoscopy was analyzed. Retrospective analysis of 120 cases of colon cancer were treated in Dongying People's Hospital from June 2015 to December 2017. All of them were treated by awakening painless digestive endoscopy, divided into 60 cases in observation group (combined with remifentanil and propofol anesthesia), and 60 cases in control group (combined intravenous anesthesia of finanib and propofol). The data were respectively recorded at time-points of oxygen inhalation, intubation for 10 min, awakening time, waking time, and the time-points for each represented as the time-points of T1, T2, T3, T4, T5 and recorded the diastolic blood pressure (DBP), respiratory rate (RR) and heart rate (HR), and compared the awakening effect and the occurrence of adverse reaction. There was no significant difference in the DBP index between the two groups at time-point T1 (P>0.05). The time-points of T2, T3, T4 and T5 were significantly different from the observation group (P<0.05). There was no significant difference in RR index between the two groups and between the same groups (P>0.05). Compared with the control group, the awakening time and consciousness recovering of the observation group is lower (P<0.05). The incidence of adverse reactions after awakening operation between the two groups was statistically significant (P<0.05). The local pain rate in the observation group after the awakening operation was lower than the control group. The combined use of trace remifentanil and small dose propofol in the awakening painless digestive endoscopy can make the patients with colon cancer more stable when they are in the awakening state, so as to improve the safety of awakening painless digestive endoscopy. It is worth promoting in clinical practice.


Characteristics of patients with hemiparetic stroke who yield highly reliable muscle strength measurements.
[Purpose] Accurate measurement of unaffected lower extremity muscle strength on the unaffected side is useful in patients with hemiparetic stroke; however, muscle strength measurement results in patients with hemiparetic stroke vary greatly compared with those in healthy individuals. The objective of the present study was to determine the characteristics of patients with hemiparetic stroke who yield highly reliable muscle strength measurements. [Subjects and Methods] The subjects were 55 incipient patients with hemiparetic stroke. Muscle strength was measured twice. Based on the measured changes and on error ranges in repeated measurements in previous studies, the subjects were divided into two groups: subjects whose measurement results were within the acceptable range, and those whose measurement results were not within the acceptable range. Logistic regression analysis was performed with this separation of groups as the dependent variable, and demographic data, physical functioning, and functional independence measure (FIM) as independent variables. [Results] From the analysis results, the FIM cognitive subscore was selected as a criterion for patient selection; the cutoff score was 19. [Conclusion] The results of the present study indicated that muscle strength measurements were highly reliable in patients with hemiparetic stroke with an FIM cognitive subscore of ≥19.


Characteristics of pesticide pyrotechnic smoke devices.
Pesticide smoke generating products are widely used by amateurs and professionals but there is little published information available about their burn and deposition characteristics to enable the risks associated with using these devices to be assessed. This paper investigates their burn characteristics, deposition patterns, pesticide air concentrations and potential exposure to operators. Thirteen firings were carried out in different spaces with different ventilation conditions. Three types of devices were investigated: dicloran, permethrin and red dye. Pesticide air concentrations increased after firing, reaching a maximum determined by the room volume in approximately 10 min and decreasing exponentially as a result of ventilation and deposition. Ejected pesticide was present in the aerosol phase but there were only occasional traces of vapour. Settlement of pesticide was affected by surface orientation, height, sampling material and the pesticide-to-space volume ratio. The manufacturer's recommended treatment period for dicloran of 4 h followed by half an hour of ventilation may be insufficient to reduce pesticide to safe levels for re-entry under very calm conditions.


Malignant melanoma. Interaction with coagulation and fibrinolysis pathways in situ.
Immunohistochemical techniques applied to fresh frozen sections of metastatic malignant melanoma tissue revealed abundant fibrinogen (or fibrin I) in perivascular areas throughout the tumor connective tissue stroma. Fibrin was readily detected in a focal distribution in the connective tissue around nodules of viable tumor. Staining for D-dimer of cross-linked fibrin (using an antibody that cross-reacted with fragment D of fibrinogen) coincided with staining for fibrin. Diffuse staining of tumor cell bodies was observed for Factor X, and Factor XIII ("a" subunit) was detected in scattered areas of connective tissue throughout the tumors. Factor VII was not detected, and only rare tumor cells stained for tissue factor. These results support the concept that a tumor cell-associated, thrombin-generating pathway exists in situ in malignant melanoma tissue that includes Factor X but neither tissue factor nor Factor VII. By contrast, tumor cell staining was observed rarely for urokinase and to a variable extent for tissue plasminogen activator.


Fibrin glue sandwich prevents pancreatic fistula following distal pancreatectomy.
Pancreatic fistula is a major form of morbidity following pancreatic resection. We conducted a nonrandomized clinical trial comparing the sealing and sandwich techniques of spraying fibrin glue to prevent pancreatic fistula following distal pancreatectomy. The pancreas was transected with a scalpel to identify and suture the main pancreatic duct and its small branches. In the sealing group, fibrin glue was sprayed over the closed pancreatic stump and sutures. Alternatively, in the sandwich group fibrin glue was sprayed so as to cover and join the cut surface of the pancreatic remnant, which was then held closed with sutures. Altogether 111 patients were included in the study (90 with gastric cancer, 10 with esophageal cancer, and 11 with pancreatic cancer). Patients were nonrandomly assigned to the sandwich or the sealing group. Morbidity was 21.8% for the patients in the sandwich group versus 33.9% in the sealing group. Pancreatic fistulas occurred in 9.0% of the sandwich group versus 26.8% of the sealing group. The incidence of fistula was thus significantly lower in the sandwich group. The incidence of fistula was also significantly lower in the sandwich group for gastric malignancy patients undergoing extended radical lymphadenectomy down to the paraaortic lymph nodes combined with left adrenalectomy. Of the patients with gastric malignancy, pancreatic fistulas occurred in 9.3% of the sandwich group versus 25.5% of the sealing group. The fibrin glue sandwich technique is simple and reliable and should be valuable for complementing other prophylactic methods of preventing pancreatic fistula.


Characterization of focal liver lesions with half-fourier acquisition single-shot turbo-spin-echo (HASTE) and inversion recovery (IR)-HASTE sequences.
The half-Fourier acquisition single-shot turbo-spin-echo (HASTE) sequence allows for heavily T2-weighted images, and the inversion recovery (IR)-HASTE sequence represents the T1 value of the tissue in a very short time. This study was undertaken to determine whether characterizing focal liver lesions can be made by combination with these very fast sequences. Seventy-four patients (33 cysts, 28 hemangiomas, and 33 malignant solid liver masses [15 metastatic tumors, 14 hepatocellular carcinomas, and 4 cholangiocarcinomas) underwent dynamic CT and breath-hold abdominal MRI using turbo-spin-echo (TSE), HASTE, and IR-HASTE sequences with variable T1 values on a 1.5-T MR unit. The imaging time for each slice was 2 seconds for HASTE imaging and 2 to 4 seconds for IR-HASTE imaging. Lesion detection and qualitative characterization were evaluated. Quantitative analysis was performed by measuring the contrast-to-noise ratios (CNRs) as well as visual analysis. The inversion time (TI) nulling values were also statistically analyzed. All cystic lesions were detected on both TSE and HASTE imagings. For solid lesions, TSE failed to detect one small solid lesion and HASTE sequence failed to detect three lesions. With HASTE sequences, all cysts and hemangiomas were markedly hyperintense in comparison with malignant solid masses. CNRs of hemangiomas or cysts were significantly higher than those of malignant solid masses (P < .01), and there was no overlap. The TI nulling value was 1,100+/-100 msec for hemangiomas, 1,900+/-110 msec for cysts, and 740+/-140 msec for malignant solid masses. There was no overlap between the TI nulling values of hemangiomas and cysts (P < .01). By combining the CNR from the HASTE sequence and the TI nulling value from the IR-HASTE sequence, complete discrimination among malignant solid masses, hemangiomas, and cysts of the liver could be made. Application of HASTE (representing T2 values) and IR-HASTE (representing T1 values) sequences provided a rapid and reliable imaging method for characterizing focal liver lesions without the use of contrast medium.


The problem of cardiac valve prostheses, anticoagulants, and pregnancy.
To assess the maternal and fetal risks in patients with cardiac valve prostheses, a series of 223 pregnancies in 156 women was studied. In 68 pregnancies (group I) treatment with antiplatelet agents was substituted for that with coumarinics from the first trimester on. Group II (128 pregnancies) received coumarinics through the thirty-eighth week of gestation. Three patients in group I died as a result of thrombosis of their caged-ball valves. The incidence of cerebral embolism was 25.0% in group I and 2.3% in group II patients, while that of spontaneous abortion was 10.3% and 28.1%, respectively. Coumarin embryopathy was found in 7.9% of group II infants. Fetal death did not occur in the 12 cases in which subcutaneous heparin was substituted for the coumarin derivative in the first trimester and the last 2 weeks of pregnancy (group III), but maternal cerebral embolism developed in 8.3% of the patients. Finally, uneventful pregnancies and deliveries were observed in 12 of 15 mothers (group IV) with biological prostheses who received no anticoagulants. It is concluded that (1) women with cardiac valve prostheses should be counseled against becoming pregnant, (2) antiplatelet agents do not provide protection against thromboembolism in these patients, (3) because of the high incidence of fetal death and birth defects use of coumarinics is contraindicated in the first trimester and the last weeks of pregnancy, (4) the substitution of heparin at these times may decrease the incidence of these complications, and (5) bioprostheses are indicated in women who wish to bear children.


Molecular analysis of ligand binding to the second cluster of complement-type repeats of the low density lipoprotein receptor-related protein. Evidence for an allosteric component in receptor-associated protein-mediated inhibition of ligand binding.
The low density lipoprotein receptor-related protein (LRP), a member of the low density lipoprotein receptor gene family, mediates the cellular uptake of a diversity of ligands. A folding chaperone, the 39-kDa receptor-associated protein (RAP) that resides in the early compartments of the secretory pathway inhibits the binding of all ligands to the receptor and may serve to prevent premature binding of ligands to the receptor during the trafficking to the cell surface. To elucidate the molecular interactions that underlie the interplay between the receptor, RAP, and the ligands, we have analyzed and delineated the binding sites of plasminogen activator inhibitor-1 (PAI-1), tissue-type plasminogen activator (t-PA).PAI-1 complexes, RAP, and the anti-LRP Fab fragment Fab A8. To that end, we have generated a series of soluble recombinant fragments spanning the second cluster of complement-type repeats (C3-C10) and the amino-terminal flanking epidermal growth factor repeat (E4) of LRP (E4-C10; amino acids 787-1165). All fragments were expressed by stably transfected baby hamster kidney cells and purified by affinity chromatography. A detailed study of ligand binding to the fragments using surface plasmon resonance revealed the presence of three distinct, Ca2+-dependent ligand binding sites in the cluster II domain (Cl-II) of LRP. t-PA.PAI-1 complexes as well as PAI-1 bind to a domain located in the amino-terminal portion of Cl-II, spanning repeats E4-C3-C7. Adjacent to this site and partially overlapping is a high affinity RAP-binding site located on repeats C5-C7. Fab A8, a pseudo-ligand of the receptor, binds to a third Ca2+-dependent binding site on repeats C8-C10 at the carboxyl-terminal end of Cl-II. Next, we studied the RAP-mediated inhibition of ligand binding to LRP and to Cl-II. As expected, we observed a strong inhibition of t-PA.PAI-1 complex and Fab A8 binding to LRP by RAP (IC50 congruent with 0.3 nM), whereas in the reverse experiment, competition of t-PA. PAI-1 complexes and Fab A8 for RAP binding to LRP could only be shown at high concentrations of competitors (>/=1 microM). Interestingly, even though the equilibrium dissociation constants for the binding of RAP to LRP and to Cl-II are similar, the binding of the ligands to Cl-II is only prevented by RAP at concentrations that are at least 2 orders of magnitude higher than those required for inhibition of ligand binding to LRP. Our results favor models that propose RAP-induced allosteric inhibition of ligand binding to LRP that may require LRP moieties that are located outside Cl-II of the receptor.


The affordability of minimally invasive procedures in major lung resection: a prospective study.
Minimally invasive procedures are used for the surgical treatment of lung cancer. Two techniques are proposed: video-assisted thoracic surgery or robotic-assisted thoracic surgery. Our goal was to study the economic impact of our long-standing program for minimally invasive procedures for major lung resection. We conducted a single-centre, 1-year prospective cost study. Patients who underwent lobectomy or segmentectomy were included. Patient characteristics and perioperative outcomes were collected. Medical supply expenses based on the microcosting method and capital depreciation were estimated. Total cost was evaluated using a national French database. One hundred twelve patients were included, 57 with and 55 without robotic assistance. More segmentectomies were performed with robotic assistance. The median length of stay was 5 days for robotic-assisted and 6 days for video-assisted procedures (P = 0.13). The duration of median chest drains (4 days, P = 0.36) and of operating room time (255 min, P = 0.55) was not significantly different between the groups. The overall conversion rate to thoracotomy was 9%, significantly higher in the video-assisted group than in the robotic group (16% vs 2%, P = 0.008). No difference was observed in postoperative complications. The cost of most robotic-assisted procedures ranged from €10 000 to €12 000 (median €10 972) and that of most video-assisted procedures ranged from €8 000 to €10 000 (median €9 637) (P = 0.007); median medical supply expenses were €3 236 and €2 818, respectively (P = 0.004). The overall mean cost of minimally invasive techniques (€11 759) was significantly lower than the mean French cost of lung resection surgical procedures (€13 424) (P = 0.001). The cost at our centre of performing minimally invasive surgical procedures appeared lower than the cost nationwide. Robotic-assisted thoracic surgery demonstrated acceptable additional costs for a long-standing program.


Crosstissue coexpression network of aging.
Aging is characterized by the interlocking decay of biological functions over time. Microarrays have been successful in elucidating some of the genome-wide changes that occur with age. Using the AGEMAP dataset that catalogs changes in gene expression as a function of age in 16 tissues in mice, we identified tissue-specific aging genes. Coordinated aging processes across different tissues then were clarified in crosstissue coexpression networks on both the gene and pathway levels. Our findings provide more concrete information about coordinated aging across different tissues. By bridging gene-level and tissue-level research, this study could help identify targets for attenuation of critical aging-related genes, pathways, or networks for antiaging intervention.


Immunoglobulin M type of autoantibodies to oxidized low-density lipoprotein has an inverse relation to carotid artery atherosclerosis.
Lipoprotein oxidation plays an important part in atherogenesis. Autoantibodies to oxidation-specific epitopes of LDL occur in plasma and atherosclerotic lesions of humans and animals. The potential role of these autoantibodies in atherogenesis still remains unsolved. We studied the relationship between different isotypes of autoantibodies to copper-oxidized LDL and malondialdehyde-modified LDL (MDA-LDL) and carotid artery intima-media thickness (IMT) in a population-based cohort of 1022 middle-aged men and women. In addition, we studied the relation of C-reactive protein (CRP) to IMT. The levels of IgM, IgG, and IgG2 autoantibodies binding to MDA-LDL and copper-oxidized LDL were determined in plasma samples by chemiluminescence-based ELISA. IMT and the number of plaques were measured ultrasonographically. The subjects were divided into tertiles for antibody titers. We found an inverse association between IMT and IgM autoantibody titers to MDA-LDL that remained statistically significant after adjusting for age, gender, LDL cholesterol, systolic blood pressure, CRP, and smoking. CRP was not independently associated with IMT. These results show that IgM autoantibodies to MDA-LDL have an inverse association with carotid atherosclerosis. The possible implications of this finding are discussed.


Bismuth oxide film: a promising room-temperature quantum spin Hall insulator.
Two-dimensional (2D) bismuth films have attracted extensive attention due to their nontrivial band topology and tunable electronic properties for achieving dissipationless transport devices. The experimental observation of quantum transport properties, however, are rather challenging, limiting their potential application in nanodevices. Here, we predict, based on first-principles calculations, an alternative 2D bismuth oxide, BiO, as an excellent topological insulator (TI), whose intrinsic bulk gap reaches up to 0.28 eV. Its nontrivial topology is confirmed by topological invariant Z 2 and time-reversal symmetry protected helical edge states. The appearance of topological phase is robust against mechanical strain and different levels of oxygen coverage in BiO. Since the BiO is naturally stable against surface oxidization and degradation, these results enrich the topological materials and present an alternative way to design topotronics devices at room temperature.


Low iodine diet for preparation for radioactive iodine therapy in differentiated thyroid carcinoma in Korea.
Preparation for radioactive iodine (RAI) therapy includes an increased serum thyroid stimulating hormone level and a low iodine diet (LID). Because of extremely high iodine intake, some physicians have advocated a more stringent LID for greater than 2 weeks in Korean patients with thyroid cancer prior to RAI therapy; however, it is very difficult to maintain a stringent LID for a longer period of time. According to recent reports in Korea, a nonstringent, simple LID for only 1 week might be enough prior to RAI therapy, if the patients can be educated intensively by specially trained staff. The measurement of simple urinary iodine concentration (UIC; µg/L) may underestimate daily iodine excretion in patients with a urinary volume of more than 1 L/day and can also be affected by dilution status. Simple UIC had a weaker correlation than the iodine/creatinine (I/Cr) ratio. Therefore, the urinary I/Cr ratio can replace 24-hour urine iodine excretion instead of simple UIC, although it may overestimate iodine intake in patients with malnutrition or poor muscle mass. The measurement of serum iodine level might be useful as an adjunct parameter for assessing LID preparation, but its sensitivity and specificity were relatively low compared to the urinary I/Cr ratio.


Sr5Ga8O3S14: A Nonlinear Optical Oxysulfide with Melilite-Derived Structure and Wide Band Gap.
Sr5Ga8O3S14, which is a novel nonlinear optical oxysulfide, was discovered via solid-state reaction. The compound crystallizes in noncentrosymmetric space group P21212 (No. 18) of the orthorhombic system. The crystal structure is derived from the melilite structure and composed of anionic [Formula: see text] layers intercalated by Sr2+ cations. The anionic layers are constructed by [GaOS3]5- and [GaS4]5- tetrahedra via either corner sharing or edge sharing, while typical melilite only features edge-sharing mode. Because of the existence of electronegative O atoms, it possesses a large band gap (3.9 eV) with a wide transparent region of 0.3-13.4 μm. It exhibits second-harmonic-generation response, which is 0.8 times that of the reference AgGaS2 (AGS) in the particle size range of 20-41 μm under laser irradiation at a wavelength of 2.09 μm laser. Density functional theory calculations indicate that the band edges are mainly composed of the Ga-S bond orbitals, with little participation of O 2p near the valence band maximum.


[Neurologic manifestations of HIV infections and AIDS].
The paper presents the review of neurological manifestations of HIV infection. The pathogenesis of changes caused by HIV within central nervous system is discussed. The most common neurological syndromes occurring in AIDS patients as well as the division of these changes concerning peripheral and central nervous system are described.


Clinical Course and Management of Vein of Galen Varix of the Neonate: A Case Report and Literature Review.
A 28-year-old pregnant woman underwent an emergency caesarian section after 39 weeks of gestation because of decreased fetal movement and baseline fetal heart rate variability. The neonate was diagnosed with neonatal asphyxia and presented with right cardiac failure due to pulmonary hypertension. The neonate presented convulsion, and plane computed tomography (CT) showed dilation of the vein of Galen and sinuses on day 3. Magnetic resonance imaging and CT with contrast were performed after cardiac failure subsided, and there was no evidence of arteriovenous shunt and normalization of the vein of Galen. The patient was diagnosed with vein of Galen varix (VGV). In the previous literature, only 3 cases of VGV have been reported. VGV is considered to be caused by right cardiac failure without the presence of an arteriovenous shunt and requires treatment only for cardiac failure and its cause. Therefore, it is important to differentiate VGV from vein of Galen aneurysmal malformation.


Psychological factors and perceptions of pain associated with dental treatment.
Although pain during dental treatment has been identified as playing a major role in the onset of dental anxiety and is a major concern of patients when seeking dental care, there have been very few studies of the prevalence of pain during dental treatment and the factors associated with patients' perceptions of pain. This study used data from a longitudinal population-based study to assess the proportion of dental attenders who experienced pain while having dental treatment and the psychological characteristics which predisposed them to experience pain. Of 1422 subjects who completed questionnaires at baseline and five-year follow-up, 96.4% had visited a dentist over the observation period. Two fifths, 42.5%, reported having pain during treatment and one-fifth, 19.1%, had pain that was moderate to severe in intensity. Reports of pain were associated with the types of treatment received, and a number of baseline sociodemographic and psychological factors. In a logistic regression analysis predicting the probability of pain, a variable documenting the number of types of invasive treatment received (restorations, extractions, crowns/bridges, root canal therapy and periodontal treatment/surgery) had the strongest independent effect. Pain was also more likely to be reported by those with previous painful experiences and those who were anxious about dental treatment, expected treatment to be painful and felt that they had little control over the treatment process. Pain was less likely to be reported by those who said they were unwilling to accept or tolerate pain. Younger subjects and those with higher levels of education were more likely to report pain than older subjects and those with a lower educational level. These results indicate that pain is as much a cognitive and emotional construct as a physiological experience. They also have implications for dentists' behaviour when providing dental care.


Novel Osmium-based Coordination Complexes as Photosensitizers for Panchromatic Photodynamic Therapy.
Cancer remains a major global malaise requiring the advent of new, efficient and low-cost treatments. Photodynamic therapy, which combines a photosensitizer and photons to produce cytotoxic reactive oxygen species, has been established as an effective cancer treatment but has yet to become mainstream. One of the main limitations has been the paucity of photosensitizers that are effective over a wide range of wavelengths, can exert their cytotoxic effects in hypoxia, are easily synthesized and produce few if any side effects. To address these shortfalls, three new osmium-based photosensitizers (TLD1822, TLD1824 and TLD1829) were synthesized and their photophysical and photobiological attributes determined. These photosensitizers are panchromatic (i.e. black absorbers), activatable from 200 to 900 nm and have strong resistance to photobleaching. In vitro studies show photodynamic therapy efficacy with both red and near-infrared light in normoxic and hypoxic conditions, which translated to good in vivo efficacy of TLD1829 in a subcutaneous murine colon cancer model.


Analysis of wound types and wound care methods after the 2023 Kahramanmaras earthquake.
This study aims to investigate the types of wounds and wound care in earthquake victims rescued from collapsed buildings after the 2023 Kahramanmaras earthquake. Between February 8th, 2023 and March 1st, 2023, a total of 94 patients (46 males, 48 females; mean age: 40.2±15.5 years; range, 16 to 77 years) with earthquake-related wounds who were trapped under rubble were retrospectively analyzed. Data including age, sex, duration of being trapped under rubble, type and location of the wound, bacterial cultures from deep tissue, and wound care methods used were recorded. The mean duration of being trapped under rubble was 58±38.1 h. Wounds were most commonly located on the lower extremities, followed by the upper extremities. The most common type of wounds were abrasions, followed by necrotic wounds due to crushing. Wound and skin antiseptics, debridement and negative pressure wound therapy (NPWT) were the most common wound care methods used. Various types of injuries and wounds may occur after natural disasters. Chronic wound care is as important as the management of life-threatening acute pathologies. Preparations should be made properly for the long-term treatment of patients after disasters. Methods such as NPWT, debridement creams containing collagenase, wound and skin antiseptics, and hyperbaric oxygen therapy can provide satisfactory short-term results. A broader and more intense application of these treatments is thought to be beneficial, particularly in crush injuries.


Acute ulnar nerve entrapment after closed reduction of a posterior fracture dislocation of the elbow: a case report.
We present the case of a child who had a posterior-lateral elbow dislocation with an intact ulnar nerve documented before an attempted reduction, with subsequent loss of ulnar nerve function after the elbow was reduced.Dislocations of the elbow in children represent only 3% to 6% of all elbow injuries. Posterior elbow dislocations are the most common, with more than one half involving an associated fracture about the elbow. The most common associated fracture is the medial epicondyle. Even though it is known that injury to the ulnar nerve can occur in elbow dislocations, we found only 1 other case report describing intra-articular entrapment of the ulnar nerve after an elbow dislocation.This case demonstrates several important clinical issues. First, it highlights the potential risk of ulnar nerve entrapment after closed reduction of elbow dislocations in children. Second, it confirms the importance of careful prereduction and postreduction examinations in the pediatric patient presenting with a dislocated elbow.


Comparing regional transcript profiles from maize primary roots under well-watered and low water potential conditions.
Regionally distinct elongation responses to water stress in the maize primary root tip have been observed in the past. A genetic basis for such differential responses has been demonstrated. Normalized bar-coded cDNA libraries were generated for four regions of the root tip, 0-3 mm (R1), 3-7 mm (R2), 7-12 mm (R3), and 12-20 mm (R4) from the root apex, and transcript profiles for these regions were sampled. This permitted a correlation between transcript nature and regional location for 15 726 expressed sequence tags (ESTs) that, in approximately equal numbers, derived from three conditions of the root: water stress (water potential: -1.6 MPa) for 5 h and for 48 h, respectively, and well watered (5 h and 48 h combined). These normalized cDNA libraries provided 6553 unigenes. An analysis of the regional representation of transcripts showed that populations were largely unaffected by water stress in R1, correlating with the maintenance of elongation rates under water stress known for R1. In contrast, transcript profiles in regions 2 and 3 diverged in well-watered and water-stressed roots. In R1, transcripts for translation and cell cycle control were prevalent. R2 was characterized by transcripts for cell wall biogenesis and cytoskeleton formation. R3 and R4 shared prevalent groups of transcripts responsible for defence mechanisms, ion transport, and biogenesis of secondary metabolites. Transcripts which were followed for 1, 6, and 48 h of water stress showed distinct region-specific changes in absolute expression and changes in regulated functions.


Generation and characterization of a monoclonal antibody against prM protein of West Nile virus.
West Nile virus (WNV), which is an emerging pathogenic flavivirus with increasing distribution worldwide, is the cause of major human and animal health concerns. The pre-membrane (prM) protein of WNV is cleaved during maturation by the furin protease into the structural protein M and a pr-segment. In this study we generated and characterized a monoclonal antibody (MAb) against the WNV prM protein. Western blot analysis showed that the MAb reacted with WNV prM specifically. Immunohistochemistry assays demonstrated that the MAb recognized native prM protein in transfected BHK-21 cells. Preliminary studies were performed to identify the epitope recognized by the MAb using a set of synthesized overlapping peptides spanning the whole length of the prM protein. The MAb reported here may provide a valuable tool for the further exploration of the biological properties and functions of the prM protein and may also be developed for potential clinical applications.


Extracellular Vesicles Derived from Wharton's Jelly Mesenchymal Stem Cells Prevent and Resolve Programmed Cell Death Mediated by Perinatal Hypoxia-Ischemia in Neuronal Cells.
Hypoxic-ischemic (HI) insult in the perinatal phase harbors a high risk of encephalopathy in the neonate. Brain cells undergo apoptosis, initiating neurodegeneration. So far, therapeutic approaches such as cooling remain limited. Transplantation of mesenchymal stem cells (MSCs) exhibits therapeutic success despite the short-time survival in the host brain, providing strong evidence that their beneficial effects are largely based on secreted factors, including extracellular vesicles (EVs). The aim of this study was to investigate the effects of human Wharton's jelly MSC (hWJ-MSC)-derived EVs on neuroprotection and neuroregeneration, using an in vitro model of oxygen-glucose deprivation/reoxygenation (OGD/R) mimicking HI injury in the mouse neuroblastoma cell line neuro2a (N2a). hWJ-MSC-derived EVs were isolated from cell culture supernatants by multistep centrifugation and identified by endosomal marker expression and electron microscopy. OGD/R significantly increased DNA fragmentation and caspase 3 ( Casp3) transcription in N2a cells relative to undamaged cells. OGD/R-mediated DNA fragmentation and Casp3 expression could be prevented as well as resolved by the addition of hWJ-MSC-derived EV before and after OGD, respectively. hWJ-MSC-derived EV also tended to increase the phosphorylation of the B cell lymphoma 2 (Bcl2) family member Bcl-2-antagonist of cell death (BAD) in N2a cells, when added prior or post OGD, thereby inactivating the proapoptotic function of BAD. Fluorescence confocal microscopy revealed the close localization of hWJ-MSC-derived EVs to the nuclei of N2a cells. Furthermore, EVs released their RNA content into the cells. The expression levels of the microRNAs (miRs) let-7a and let-7e, known regulators of Casp3, were inversely correlated to Casp3. Our data suggest that hWJ-MSC-derived EVs have the potential to prevent and resolve HI-induced apoptosis in neuronal cells in the immature neonatal brain. Their antiapoptotic effect seems to be mediated by the transfer of EV-derived let-7-5p miR.


Insulin and rabbit anti-insulin receptor antibodies stimulate additively the intrinsic receptor kinase activity.
This paper describes the properties of rabbit polyclonal antibodies directed against purified human insulin receptor which strongly stimulate the intrinsic tyrosine kinase activity. The stimulatory effect of the antibodies on the kinase activity was obtained on the insulin receptor autophosphorylation as well as on the kinase activity towards a synthetic substrate. This stimulation is additive to that induced by insulin. Moreover, rabbit antibodies do not impair insulin binding. These data strongly suggest that antibodies and insulin act through separate pathways. This conclusion is reinforced by the differences observed on the phosphopeptide maps of the receptor's beta subunit whose phosphorylation was performed either in the presence of insulin or rabbit antibodies. Interestingly, these polyclonal antibodies can also induce an activation of the receptor autophosphorylation by interacting only with extracellular determinants. The anti-insulin receptor antibodies mimic insulin in their stimulatory effect on amino acid (AIB) uptake, but they have a different effect to that found on the kinase activity; the simultaneous addition of the antiserum and insulin failed to stimulate this amino acid transport over the level induced by a saturating concentration of hormone.


Optimization of dilute acid pretreatment of water hyacinth biomass for enzymatic hydrolysis and ethanol production.
The present study was conducted for the optimization of pretreatment process that was used for enzymatic hydrolysis of lignocellulosic biomass (Water Hyacinth, WH), which is a renewable resource for the production of bioethanol with decentralized availability. Response surface methodology has been employed for the optimization of temperature ((o)C), time (hr) and different concentrations of maleic acid (MA), sulfuric acid (SA) and phosphoric acid (PA) that seemed to be significant variables with P < 0.05. High F and R (2) values and low P-value for hydrolysis yield indicated the model predictability. The pretreated biomass producing 39.96 g/l, 39.86 g/l and 37.9 g/l of reducing sugars during enzymatic hydrolysis with yield 79.93, 78.71 and 75.9 % from PA, MA and SA treated respectively. The order of catalytic effectiveness for hydrolysis yield was found to be phosphoric acid > maleic acid > sulfuric acid. Mixture of sugars was obtained during dilute acid pretreatment with glucose being the most prominent sugar while pure glucose was obtained during enzymatic hydrolysis. The resulting sugars, obtained during enzymatic hydrolysis were finally fermented to ethanol, with yield 0.484 g/g of reducing sugars which is 95 % of theoretical yield (0.51 g/g glucose) by using commercial baker's yeast (Sacchromyces cerveasiae).


[Emergency surgical treatment of complicated acute diverticulitis].
Twenty-five percent of patients undergoing surgery for acute complicated diverticulitis represent emergencies. This condition is currently treated by colonic resection with primary anastomosis with or without colostomy, or by a Hartmann operation. We report on our experience with 52 consecutive patients with generalized peritonitis (8 cases), peri- and paracolonic abscesses (19 cases), severe pelvic abscesses (12 cases) and multiple abscesses with visceral fistulas (13 cases). All patients had emergency surgery. In 50/52 patients (96.2%) we performed a colonic resection with primary anastomosis using a mechanical stapler and in 2/52 a Hartmann operation. The overall mortality rate was 5.8%. The morbidity rate was 22% with 9 anastomotic leakages. A diverting colostomy was constructed in 16 patients and opened in only 8 patients. In 4 cases a parastomal hernia occurred after late closure and reduction of the colostomy. This data suggest that colonic resection with primary anastomosis, even without colostomy, is a safe procedure for the emergency treatment of acute complicated diverticulitis.


[The last outbreak of smallpox in Sweden].
On April 6 1963 a fairly well vaccinated seaman, newly arrived to Sweden from the Far East, developed a mild smallpox. He caused an outbreak of smallpox involving 26 further cases. This article describes how the illness was spread in Stockholm.


Medial Plantar Artery Perforator Flap: Experience with Soft-tissue Coverage of Heel.
Soft-tissue coverage is a challenge to the ankle and foot reconstructive surgeon due to its unique requirement of simultaneously withstanding body weight and to provide sensory feedback. We share our experience of medial plantar artery perforator (MPAP) flap, which provides a robust sensate coverage to heel defects. Three-year retrospective study, which included soft-tissue injury to heel. All patients underwent MPAP flap for the coverage of the defect. Patients' demographic, mode of injury, defect size, flap size and survival, time to start weight bearing, return of protective sensations, and comparative 2-point discrimination with opposite heel were studied. We studied 16 cases with heel soft-tissue injuries. Fifteen had motor vehicle accident, and 1 had chronic diabetic wound. Mean surface area of flap was 4 × 5 cm. Except 1 flap, 15 flaps had complete survival and provided reliable wound coverage with comparable normal sensation as on other foot. We suggest that MPAP flap, when available, is a good local tissue for heel wounds coverage.


Effects of induction of rRNA overproduction on ribosomal protein synthesis and ribosome subunit assembly in Escherichia coli.
Overproduction of rRNA was artificially induced in Escherichia coli cells to test whether the synthesis of ribosomal protein (r-protein) is normally repressed by feedback regulation. When rRNA was overproduced more than twofold from a hybrid plasmid carrying the rrnB operon fused to the lambda pL promoter (pL-rrnB), synthesis of individual r-proteins increased by an average of about 60%. This demonstrates that the synthesis of r-proteins is repressed under normal conditions. The increase of r-protein production, however, for unknown reasons, was not as great as the increase in rRNA synthesis and resulted in an imbalance between the amounts of rRNA and r-protein synthesis. Therefore, only a small (less than 20%) increase in the synthesis of complete 30S and 50S ribosome subunits was detected, and a considerable fraction of the excess rRNA was degraded. Lack of complete cooperativity in the assembly of ribosome subunits in vivo is discussed as a possible explanation for the absence of a large stimulation of ribosome synthesis observed under these conditions. In addition to the induction of intact rRNA overproduction from the pL-rrnB operon, the effects of unbalanced overproduction of each of the two large rRNAs, 16S rRNA and 23S rRNA, on r-protein synthesis were examined using pL-rrnB derivatives carrying a large deletion in either the 23S rRNA gene or the 16S rRNA gene. Operon-specific derepression after 23S or 16S rRNA overproduction correlated with the overproduction of rRNA containing the target site for the operon-specific repressor r-protein. These results are discussed to explain the apparent coupling of the assembly of one ribosomal subunit with that of the other which was observed in earlier studies on conditionally lethal mutants with defects in ribosome assembly.


Relation of lymphoid system and hormones to aging.
Congenital deficiencies of developmental hormones, such as growth hormone and thyroxine, are responsible for the thymus-dependent immunodeficiency disease observed in dwarf mice. Such an immunodeficiency state is associated with early aging phenomena. This fact suggests that lymphoid cells and primarily T-derived cells are involved in the control of aging processes.


Evaluation of the relationship between salivary concentration of anti-heat shock protein immunoglobulin and clinical manifestations of Behçet's disease.
This study investigated the relationship between the concentration of anti-heat shock protein (HSP) 60 antibody in resting saliva and the severity of Behçet's disease (BD). Sixty-five patients diagnosed with BD at Tokyo Medical and Dental University Hospital were enrolled in this study. Based on clinical severity scores, patients were categorized as having mild, moderate, or severe BD. Periodontal status was evaluated with the Community Periodontal Index (CPI), and anti-HSP60 antibody concentrations in resting saliva were measured with an enzyme-linked immunosorbent assay. The mean antibody concentration in patients in the moderate group was significantly higher than concentrations in the mild and severe groups. No significant difference was found between the mild and severe groups. Gingival inflammation, identified with the CPI, was associated with a higher antibody concentration. The antibody concentration in patients who had stomatitis for more than 2 weeks was significantly higher than in those with stomatitis for less than 2 weeks. The antibody concentration in patients who had taken colchicine was significantly lower than that in subjects who had not. These results suggest that the concentration of anti-HSP60 antibody in resting saliva may be effective as a non-invasive indicator for the diagnosis (screening) and prognostication of BD.


Innate Immune Memory: Implications for Microglial Function and Neuroprogression.
Immunostimulatory insults such as stress and infection are risk factors for the development of several neuropsychiatric disorders characterized by neuroprogression. Inflammatory and neurotoxic molecules in the brain can cause disruptions in neurogenesis, neuronal excitability, synaptic transmission, synaptic plasticity, and neuronal survival - changes that characterize neuroprogression. We draw on recent findings in the immunology literature that peripheral innate immune cells are capable of retaining long-term memory of infectious insults and displaying long-lasting upregulated proinflammatory function in response to repeated infectious insults - a concept known as "innate immune memory." In turn, we hypothesize that microglia, the resident innate immune cells of the brain, are also capable of retaining long-term memory of infectious and noninfectious insults, including stress. Microglia are capable of producing a variety of proinflammatory neurotoxic cytokines and chemokines. Persistent upregulation of microglial proinflammatory function as a result of memory for immunostimulatory insults may therefore contribute to persistent and progressive inflammation in neuropsychiatric illnesses and be an important driver of neuroprogression.


The impact on patients of a nurse-led clinical service in gastroenterology.
Higher rates of chronic liver disease have resulted in a significant increase in the number of patients needing regular abdominal paracentesis for ascites. Waiting times for admission at the Royal Wolverhampton NHS Trust had become longer and delays in intervention and treatment became inevitable. In 2014, the Trust developed a nurse-led abdominal paracentesis day-case service. A qualitative evaluation of the service was conducted using in-depth patient interviews and surveys to determine the impact of the service, including how the new nursing role affected the patient experience. The results show that the nurse-led clinical service in gastroenterology positively impacts patients' experiences. Following this innovation, the service was extended to provide intravenous infusions for gastroenterology patients via the day-case unit. With appropriate training and competency assessments, nurses can now perform selected medical procedures safely and effectively, thereby facilitating nursing staff to expand and develop their roles. This development has substantial implications for nursing and is an important contribution to the debate on the future direction of the nursing profession.


Differential reduction in soluble and membrane-bound c-type cytochrome contents in a Paracoccus denitrificans mutant partially deficient in 5-aminolevulinate synthase activity.
A mutant of Paracoccus denitrificans, DP104, unable to grow anaerobically with nitrate as the terminal electron acceptor or aerobically with methanol as the electron donor and staining negatively in the dimethylphenylene diamine oxidation (Nadi) test, was isolated by transposon Tn5::phoA mutagenesis. P. denitrificans DP104 grown aerobically with succinate or choline had very low levels (2 to 3% of the wild-type levels) of spectroscopically detectable soluble c-type cytochromes. In contrast, membrane cytochromes of the a, b, and c types were present at 50% of the levels found in the wild type. The apo form of cytochrome c550, at an approximately 1:1 molar ratio with the holo form, was found in the periplasm of DP104. The TnphoA element was shown to be inserted immediately upstream of the translational start of hemA, the gene coding for 5-aminolevulinate synthase, which was sequenced. Low-level expression of this gene, driven off an incidental promoter provided by TnphoA-cointegrated suicide vector DNA, is the basis of the phenotype which could be complemented by the addition of 5-aminolevulinate to growth media. Disruption of the hemA gene generated a P. denitrificans strain auxotrophic for 5-aminolevulinate, establishing that there is no hemA-independent pathway of heme synthesis in this organism. The differential deficiency in periplasmic c-type cytochromes relative to membrane cytochromes in DP104 is suggested to arise from unequal competition for the restricted supply of heme which results from the effects of the transposon insertion.


Amino acid sequence homology among the major outer membrane proteins of Escherichia coli.
Analysis of amino acid sequences reported for the major outer membrane proteins of Escherichia coli, including the porins (OmpF, OmpC, and PhoE), the phage lambda receptor (LamB), and another protein (OmpA), revealed several regions of local homology that is statistically significant. The implications of this observation are discussed in relation to the evolutionary origins of these proteins, as well as to the mechanism of export of these proteins to the outer membrane.


Long-term follow-up after gentamicin application via the Silverstein MicroWick in the treatment of Ménière's disease.
We conducted a retrospective study to evaluate the long-term effectiveness of gentamicin applied directly to the round window membrane via the Silverstein MicroWick system in patients with definite or probable Ménière's disease. The study population was made up of 69 patients who had received 3 drops of gentamicin 10 mg/ml three times daily; the duration of treatment was guided by objective data obtained from weekly electronystagmography and audiometry. Relief of vertigo was assessed by chart review and telephone interviews. After a minimum follow-up of 24 months, we found that vertiginous symptoms remained controlled in 53 patients (76.8%) after one or more courses of drug therapy. A single course of treatmentwas successful in 41 patients (59.4%). We conclude that long-term control of vertigo can be achieved inpatients with Ménère's disease by direct application of gentamicin to the round window membrane via the Silverstein MicroWick. This minimally invasive self-treatment technique was well tolerated and free of long-term complications.


Fluorescent Labeling of Yeast Cell Wall Components.
Yeast cells stained with a fluorescent dye that specifically binds to one of the cell wall components can be observed under a fluorescent microscope. Visualization of the components 1,3-β-glucan, mannoproteins, and/or chitin not only provides information concerning the cell wall, but also reveals clues about various cellular activities such as cell polarity, vesicular transport, establishment of budding pattern, apical and isotropic bud growth, and replicative cell age. This protocol describes a standard method for visualizing different components of the yeast cell wall.