[in Japanese]

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Serotypes and distribution of the virulence genes of Escherichia coli isolates from healthy individuals were investigated. Four-hundred and 38 E. coli strains, from 428 (9.2%) of 4,667 individuals, were serotypable by using of the commercially available serotyping serum, and those E. coli strains were classified into 37 O serotypes, 20 H and 118 OH serotypes. The most frequent O serotype was O1 (32.3%), followed by O18 (15.9%). The virulence genes (it, st, stx, eaeA, invE, aggR, and astA) were detected in 75 (17.1%) out of 438 strains. Only two strains of O1 and O18 serotypes had the virulence genes. In antibiotic sensitivity tests for 12 drugs, 29 diarrheagenic E. coli strains were resistant, especially all 24 strains harboring aggR were resistant to ABPC. It is risky that the screening by serotyping can also cover non-pathogenic E. coli from our results. The precise screening for the virulence factors by PCR after the screening by serotyping against the diarrheagenic E. coli isolates is important. For some virulence factors, eaeA and astA, which are uncertain in the relationship with serotypes, I should make clear their distribution pattern and the relationship with serotypes in the diarrheagenic E. coli isolates in future.

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Background: Steroid preparation exerts its pharmacological effect through the binding of the free unbound fraction to its receptor. However, in patients with nephrotic syndrome, the plasma protein concentration fluctuates, which affects the ratio of bound versus unbound steroid. We investigated the plasma pharmacokinetics of prednisolone administered to patients with nephrotic syndrome through their disease course. Also the suppression of endogenous cortisol during prednisolone therapy was studied. Methods: Fifteen patients (aged 16-75 years, nine minimal change group, four membranous nephropathy, two lupus nephritis) were studied on one or more occasions over a period of one to nine months during prednisolone treatment. Plasma total concentrations of prednisolone were measured by employing gas chromatography/mass spectrometry simultaneously with endogenous cortisol at several time points on each occasion. Individualized estimation of the patient's kinetic parameters was done by a computer curve fitting of the total plasma prednisolone. Results: Blood sampling at several time points on each occasion made it possible to characterize the intra- and inter-individual variability in the pharmacokinetic behavior of prednisolone. There was a negative correlation (r=-0.4676, P=0.0031) between the prednisolone clearance and plasma albumin concentrations. As the plasma albumin level increases, the area under the plasma total prednisolone concentration-time curve (AUC_<total>) increased, whereas the area under the plasma unbound prednisolone concentration-time curve (AUC_<unbound>) decreased. The discrepancy between AUC_<total> and AUC_<unbound> was observed at the plasma albumin concentration below 3.0 g/dl. The endogenous cortisol concentration immediately declined after administering the first prednisolone dose. Adrenal suppression is less in patients in whom prednisolone was administered only in the morning than in patients in whom prednisolone was administered in two or three divided doses. Conclusion: Concentration of free unbound prednisolone decreases as the plasma albumin concentration is recovered. It is recommended that the dosage reduction be started after the plasma albumin concentration is normalized and stabilized (above 3.0 g/dl). In addition administration of prednisolone once daily in the morning is recommended to prevent adrenal suppression.

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To investigate how proportion of each occurrence of the four CHD risk factors (hypertension, high total cholesterol level, low HDL-cholesterol level, and diabetes mellitus) or clustering of these factors can be attributed to obesity (25 or greater of body mass index (BMI)), the cross-sectional health examination data for 582 male workers of a sales company in Tokyo metropolitan was statistically analyzed according to four subgroups crossed by two age categories (45yrs or more and 44yrs or less) and two smoking categories (non-smokers and current smokers). The results showed that the contribution of obesity to each occurrence of the four risk factors was greater in younger group than in older group in non-smokers. However, in smokers, the contribution of obesity to clustering of the four risk factors was remarkably large irrespective of age. It was estimated that about half of the health status with clustering of these factors can be attributed to obesity for current smokers in males.

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The pathogenetic process of the keloid or hypertrophic scar formation is still unknown. To make the mechanism of the scar formation clearer, gene expression levels of fibroblasts derived from hypertrophic scar were estimated and compared with those prepared from healthy skin of the same individual by the gene expression array. Fibroblasts obtained from five post-burn patients under the informed consent were cultured for several passages. The gene expression levels in those cultured cells were analyzed by a Nylon-membrane array that was hybridized with [33P]-labeled cDNA probes prepared from mRNA obtained from those cells. In a total of 1,179 genes on the array 18 genes exhibited a statistically significant difference at a level of p<0.05 and another 13 genes at the level of 0.05<p<0.1 in the expression between the two cultured fibroblasts by the student's t-test The expression levels of several genes related to cell-growth regulation, such as growth factors, G-proteins and cellular kinase networks, were lower in cultured fibroblasts obtained from hypertrophic scar than those in healthy skin cells. Apoptosis-related genes, BNIP3, DAXX, RXRB, APAF1, BMP3B, PRL1, ACE, and HOXA5 changed their expression levels in the scar cells. Genes for cell cycle-regulation associated factors, CDK4I, STAB1, cycline K, and cycline A showed a depressed expression in the scar fibroblasts. Those alterations in the gene expression will influence on the cellular signaling pathways and will be involved in the scar formation.

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In this study the predict values of several factors that may influence on testicular sperm retrieval rate were evaluated in patients with azoospermia undergoing testicular sperm extraction (TESE). Serum levels of FSH, LH, prolactin (PRL), testosterone (T), dehydrotestosterone (DHT), estradiol (E_2), 17 α-hydroxy-progesterone were measured and cross sectional area of both side testes was measured by ultrasonography in 43 patients with azoospermia. Testicular sperm retrieval rate was significantly decreased in the group caused by idiopathic testicular dysfunction compared to those by obstructive, cryptorchism, and ejaculatory dysfunction (p<0.05). Cross sectional area of both side testes had no predictive value for testicular sperm retrieval rate. In patients whose FSH levels were ≦ 15mIU/ml or LH levels were ≦ 2mIU/ml showed 100% sperm retrieval rate. In contrast, no sperm was retrieved in patients whose FSH levels were >60mIU/ml or LH levels were >12mIU/ml. In patients whose levels of FSH and LH were between above two group, testicular sperm retrieval rate was 33% in which sperm retrieval rate was 58% and 0 % in patients with DHT levels ≦ 0.5ng/ml and >0.5ng/ml, respectively. These results indicate that measurement of FSH, LH and DHT is useful to predict the testicular sperm retrieval rate by which harmful and invasive TESE can be avoided.

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The aim of this study is to evaluate the effects of chronic administration of scopolamine or diazepam on spatial learning in rats. Animals were trained at 28 days (1 trial per day), and scopolamine or diazepam was administered 30 min before each trial. At the results, control group increased win-shift behavior, and decreased reference error. But, scopolamine and diazepam groups did not induce to decrease reference error compared with control group. Working error was increased by scopolamine administration, but not diazepam. In withdrawal trials, diazepam withdrawal group significantly decreased win-shift behavior compare with diazepam group. These results suggest the differential mechanism between scopolamine and diazepam on spatial learning in rats and indicate that acetylcholine and benzodiazepine receptor plays the important role of spatial learning formation.

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The aim of this study is to evaluate the role of NMDA receptor/nNOS pathway on passive avoidance learning and memory processes in rats, and the effect of DHEAS, as a neurosteroid, on learning and memory. Eight weeks old Wistar-Imamichi rat were used, and passive avoidance learning and memory was evaluated with the Step-through type passive avoidance apparatus. In results, MK-801, NMDA receptor antagonist, significantly impaired acquisition and retrieval in dose-dependent manner. Selective nNOS inhibitor 7-NI impaired the acquisition performance, and mildly impaired retrieval performance. MK-801-induced acquisition impairment was reversed by pre-treatment of DHEAS 20mg/kg administration, significantly, but same doses of DHEAS administration did not prevent MK-801-induced retrieval impairment and 7-NI-induced acquisition deficit. Furthermore, All drugs did not influence retention performance. These results suggest that NMDA receptor/nNOS pathway may play an important role in the acquisition phase of passive avoidance learning and memory in rats. Moreover, the effects of DHEAS on learning deficits were appeared through NMDA receptors, and nNOS was not related to the effects of DHEAS.

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Acellular allogeneic dermal matrix (ADM) has been used for simultaneous transplantation with autologous skin graft overlay and a scaffold of cultured composite skin. The effectiveness of ADM transplantation for the dermal regeneration has been widely accepted. However, possibility of disease transmission by ADM remains as a potential risk. So it is important to reduce the risk by ADM transplantation. The purpose of this study was to evaluate whether ADM could be disinfected and sterilized without losing its transplantation properties. Four different methods of disinfection and sterilization, such as 98% glycerol, pasteurization, gamma irradiation and ethylene oxide gas (EOG) were used in this study. After disinfecting and sterilizing, histological, immunohistochemical (anti-type IV collagen and laminin) and electrophoretic (SDS- PAGE) properties were evaluated. The property of simultaneous transplantation with skin graft overlay was studied using a rat model. Finally, the property as a scaffold of cultured tissue engineered skin (cultured composite skin) was evaluated using normal human cultured keratinocytes. All the disinfection and sterilization methods did not induce apparent collagen degradation of ADM on SDS-PAGE. However histological damages of basement membrane structure (type IV collagen and laminin) were observed by ADMs treated with Pasteurization, Gamma irradiation, and EOG. The survival rate of overlaid skin grafts was significantly reduced when Pasteurization, Gamma irradiation, and EOG treated ADM was simultaneously transplanted. These three methods also resulted in poor regeneration of epidermal layer in cultured composite skin. Through all of the evaluation, glycerol treatment was found to be the best way of ADM sterilization as far as its transplantation property was concerned.

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Concentrations of VEGF in EMF was significantly higher in patients with endometrial carcinoma compared to those with myomas, adenomyosis, endometrial hyperplasia, and normal control. In patients with endometrial carcinoma, concentrations of VEGF in EMF increased from the stage 0 to Ib, reaching a plateau at stage Ib.Levels of VEGF in EMF from patients with endometrial carcioma did not differ between the grades G1 and G2. VEGF levels in EMF are higher in patients with muscularinvasion, compared to those without invasion. However, neither the presence of vascular invasion nor metastasis of lymphnode influenced on levels of VEGF in EMF. Multiple logistic regression showed that among VEGF in EMF, serum CA125 and MRI staging, VEGF in EMF alone was a significant predictor for the surgical stage Ib or more. Discriminant analysis using 3 parameters of VEGF in EMF, serum CA125 and age predicted the surgical stage Ib or more with 0.94 of sensitivity and 0.86 of specificity.

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Surgery for patients with chronic liver dysfunction carries a high risk of postoperative complications including liver failure. In these patients, indications for surgery should depend on assessment of hepatic functional reserve. Furthermore, careful perioperative management is required. We herein report a case which died of postoperative liver failure despite the indication and management based on such strategy. A 65-year-old man with alcoholic liver disease was found to have intraductal papillary-mucinous tumor of the pancreas on imaging diagnosis. Preoperatively, he had good hepatic functional reserve (Child A). He underwent pylorus-preserving pancreatoduodenectomy. Despite careful perioperative management, he developed liver failure and died three days later. Autopsy revealed acute necrosis superimposed on the enlarged cirrhotic liver. A safer strategy should be established for surgical treatment in patients with chronic liver dysfunction.

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