Existence of early-onset NIDDM Japanese demonstrating severe diabetic complications.

OBJECTIVE: To identify the clinical characteristics of early-onset NIDDM patients with severe diabetic complications. RESEARCH DESIGN AND METHODS: The clinical cases of a large number of diabetic patients who visited a diabetes center within the period 1970-1990 were reviewed. Of a total of 16,842 diabetic patients, 1,065 (6.3%) had early-onset NIDDM (diabetes diagnosed before 30 years of age). These 1,065 patients were divided into two groups, those who developed proliferative retinopathy before the age of 35 (n = 135) and those who did not (n = 930). Development of proliferative retinopathy, nephropathy, renal failure, blindness, and atherosclerotic vascular disease were compared between the two groups. RESULTS: The subgroup of 135 patients was characterized by poor glycemic control, often requiring insulin therapy and a higher familial prevalence of diabetes, and contained a greater proportion of women than the subgroup of 930 patients. Of the 135 patients, 99 (67%) developed proliferative retinopathy before the first visit. The 135 patients developed severe progressive complications in contrast to the 930 patients. A total of 81 patients (60%) developed diabetic nephropathy at a mean age of 31 years, 31 (23%) developed renal failure requiring dialysis at a mean age of 35 years, 32 (24%) became blind at a mean age of 32 years, and 14 (10%) developed atherosclerotic vascular disease at a mean age of 36 years. CONCLUSIONS: Some Japanese early-onset NIDDM patients develop severe diabetic complications in their youth. Most of them had no symptoms nor regular treatment regarding diabetes until they were noticed to have developed severe diabetic complications. Although the relevant prevalence and the pathogenetic mechanism underlying the rapid onset of the complications remain to be determined, prolonged inadequate treatment of and familial predisposition to diabetes may be contributing factors. Careful diabetes care in the twenties, not only for IDDM but also for NIDDM patients, is warranted.

High incidence of diabetic nephropathy in early-onset Japanese NIDDM patients. Risk analysis.

OBJECTIVE: Because early-onset Japanese NIDDM patients (diagnosed before age 30 years) can develop diabetic end-stage renal failure (ESRF) in their thirties, this study was performed to elucidate the incidence and determinants for the development of diabetic nephropathy. RESEARCH DESIGN AND METHODS: The incidence of diabetic nephropathy and its relationship to baseline characteristics and long-term metabolic control were determined in 426 early-onset Japanese NIDDM patients who were followed for a mean of 6.8 years. RESULTS: Of these 426 patients, 41 developed diabetic nephropathy manifested by persistent proteinuria (incidence rate [95%CI]/1,000 person-years; 14.1 [10.4-19.1]). Among patients whose mean HbA1c (measured by a high-performance chromatography method that is standardized and comparable to the one used in the Diabetes Control and Complications Trial study) was around 7% or less, few developed nephropathy. The incidence of nephropathy increased with increasing mean HbA1c level in a dose-dependent manner (chi 2 trend = 49.9, P < 0.0001). Diastolic blood pressure and duration of diabetes at entry had significant predictive effects independent of metabolic control. CONCLUSIONS: The incidence rate of diabetic nephropathy in early-onset Japanese NIDDM patients is potentially high, similar to or higher than that in Pima Indian NIDDM or Caucasian IDDM patients of comparable age. Diabetic nephropathy in NIDDM patients aged in their thirties or forties is likely to be an early feature that leads to ESRF, and this would contribute to the marked increase in the number of new patients with diabetic ESRF in Japan. NIDDM is a serious disease if near-normal glycemia is not achieved.

Biosynthesis of L-alanine, a major amino acid of fibroin in Samia cynthia ricini.

The derivation of alanine in fibroin was investigated using NMR and selective isotopic labelling. 2H2O infused orally into 5th instar larvae was incorporated into the proton of the methyl group of alanine in fibroin. Proton exchange among alanine, glycine and serine was also found. Incorporation of 13C from [2-(13)C]acetate into alanine C2 and C3 and glycine C2 in fibroin, and also C4 of free glutamine plus glutamate was observed in vivo. Hemolymph contained a peak for C4 of glutamate plus glutamine, and an alanine C3 peak appeared transiently. Thus, it is suggested that the C-skeleton of alanine formed was derived from L-malate via the TCA-cycle, and that this alanine is utilized in part for fibroin synthesis. Spectra of the hemolymph extract of larvae infused orally with [15N2]urea showed no 15N-compounds, whereas those of larvae injected subcutaneously showed only one peak of urea, whose intensity decreased with time, as shown in the in vivo spectra of a living larva infused with [15N2]urea. The solution NMR spectrum of fibroin showed no 15N-labelled compounds. Temporal changes in the peak intensities of six compounds in the spectra of a living larva infused with [15N]ammonium demonstrated a process in which 15N was incorporated into fibroin containing 15N-alanine through the amide group of glutamine and the amino group of glutamate. Thus, alanine biosynthesis from the TCA-cycle originates mainly from water, L-malate and ammonium. The fact that no 15N-urea was detected in the hemolymph extract of larvae infused with [15N]ammonium suggests that 15N-urea found in the above in vivo spectra may be that accumulated in the hindgut. Thus, excess ammonium in the body causes the production of urea by the urea-cycle. In Samia larvae, urea was not reutilized but excreted. The metabolic relationships between the assimilation of ammonium and the function of the urea-cycle are discussed.

Molecular-genetic analysis of ocular adnexal benign lymphoid hyperplasias by a two-step polymerase-chain-reaction.

Twelve biopsied ocular adnexal benign lymphoid hyperplasias (OABLH) satisfying benign histological criteria were investigated for clonal immunoglobulin (Ig) heavy-chain gene rearrangement by means of a two-step polymerase chain reaction (PCR) method using formalin-fixed and paraffin-embedded tissue. Of the 12, 4 (33%) demonstrated clear single bands of the rearranged gene for the Ig heavy-chain, of between 100 and 150 base pairs. The selected cases were all free of malignant lymphoma and all of the lesions were small (2 x 2-22 x 6 mm; median 4.5 x 3 mm). Histopathological and cytological features were not essentially different between monoclonal and non-monoclonal examples. Immunohistochemistry was of little benefit in separating the two. It is concluded that OABLH demonstrating a benign clinical course frequently contain monoclonal B cell populations suggesting a continuous progressive spectrum of lesions in B cell neoplasia. In addition, the significance of molecular-genetic analysis for OABLH and the utility of the two-step PCR method should be emphasized.

Argyrophilic nucleolar organizer regions in human adrenocortical neoplasms.

Argyrophilic nucleolar organizer regions (AgNOR) in human adrenocortical neoplasms, including five carcinomas and ten adenomas, were studied using a semi-automatic image analyzer. Both the number and total area of AgNOR per nucleus in the carcinomas were found to be statistically greater than in adenomas and control tissues. However, there were no statistically significant differences in total AgNOR area per nuclear area or in the mean area of individual AgNOR dots. The AgNOR of neoplastic and normal cells were of four morphological types: type 1 had a few dots at the periphery of the nucleus, type 2 a few dots at the center, type 3 a large round dot along with several small ones at the center, and type 4 numerous diffusely distributed polymorphic dots. Most type 3 and 4 cells were found in carcinoma cases. Type 1 cells decreased in proportion to the severity of biological malignancy. It follows from these findings that careful observation of AgNOR should facilitate the distinction of malignant from benign adrenocortical neoplasms.

A possible role for Epstein-Barr virus in tumorigenesis after immunosuppression in cases of renal transplantation.

Eight secondary malignancies developing after renal transplantation were investigated in terms of a possible role of the Epstein-Barr virus (EBV). In five cases, four gastric cancers and one colonic cancer, the presence of EBV was proven by the polymerase chain reaction (PCR), all four gastric lesions being confirmed to have a massive EBV infection by in situ hybridization. Two cases demonstrated monoclonal infection with EBV, as indicated by a single band of the lymphocyte-defined membrane antigen tandem-repeat gene using PCR, and were immunohistochemically positive for the latent membrane protein 1. Our series suggests that gastrointestinal cancer predominates as a secondary malignancy in states of induced severe immunosuppression, and that EBV may play an important role in tumorigenesis as an oncovirus.

Comparative analysis of p53 and c-myc expression and cell proliferation in human hepatocellular carcinomas--an enhanced immunohistochemical approach.

Expression of p53 and c-myc was investigated and compared with cell proliferative activity in a series of 40 hepatocellular carcinomas (HCC), by means of enhanced immunohistochemistry. p53 expression was demonstrated in 5 out of 40 HCC (12.5%) with the incidence increasing in 5 out of 40 HCC (12.5%) with the incidence increasing in proportion to the histological grading of malignancy: thus, 0% of well-differentiated, 6.9% of moderately differentiated and 33.3% of poorly differentiated lesions were positive. The proliferating-cell nuclear antigen (PCNA) labeling index also showed a statistically significant increase with this grading. Distribution patterns of PCNA-positive cell were divided into four types: scatter, marginal, mosaic and diffuse. Four HCC cases, predominantly of the poorly differentiated type, exhibited the diffuse pattern. Generally, p53 overexpression corresponded well with PCNA positivity. In contrast, there was no correlation between c-myc overexpression, found in 19 out of 40 HCC (47.5%), and histological grading of HCC or PCNA labeling index. The distribution pattern of c-myc-positive HCC cells was also different from that of PCNA and p53. Our results suggest that p53 overexpression closely relates to proliferation of HCC cells. Furthermore, there may be a consistent difference in regulatory mechanisms between p53 and c-myc expression in multistep hepatocarcinogenesis.

Slightly elevated blood pressure as well as poor metabolic control are risk factors for the progression of retinopathy in early-onset Japanese Type 2 diabetes.

Not a few patients in Japan with early-onset type 2 (non-insulin-dependent) diabetes become blind due to proliferative diabetic retinopathy (PDR). However, the risk factors are poorly understood. The aim of this study was to determine the risk factors for background diabetic retinopathy (BDR) and PDR by following 394 Japanese patients with early-onset type 2 diabetes diagnosed before 30 years of age (mean age 27, mean blood pressure at entry 116/73 mm Hg). Of the 322 patients who were free of diabetic retinopathy at entry, 88 developed BDR, giving an incidence of 57.7 (95% CI 55.5-60. 0)/1000 person-years. Cox proportional hazard analysis revealed mean HbA(1c) and duration of diabetes to be significant predictors of development of BDR. Of the 160 patients with BDR, i.e., the 72 patients who had BDR at entry and the 88 who developed BDR during the follow-up, 50 developed PDR, giving an incidence of 17.9 (95% CI 13.6-23.6)/1000 person-years. Cox proportional hazard analysis indicated mean HbA(1c) and diastolic blood pressure to be significant predictors of the progression from BDR to PDR. In conclusion, in early-onset Japanese type 2 diabetic patients, the rates of both development of BDR and of progression from BDR to PDR appear to be potentially high. Not only lifetime exposure to glycemia but also a slightly elevated blood pressure level is an important risk factor for progression to PDR.

Hepatic histopathologic range compared with virological studies of hepatitis viruses among autopsy cases in Tokyo.

Few reports exist comparing virological studies on hepatitis viruses with histopathological studies of autopsy cases other than those of liver clinics. Relations between hepatitis virus-related markers and hepatic histopathology were studied in 1044 autopsy cases (779 men and 265 women) at the Medical Examiner's Office, Tokyo. Heart blood was obtained at the autopsy, and the sera were submitted for virus-marker detection of HBV, HCV, and HGV/GBV-C. Hematoxylin and eosin-stained paraffin sections were used for histological assessment. Histopathologically, 463 cases were determined as so-called normal liver; among them 440 cases (95.0%) were negative for all hepatitis virus-related markers, but HBV-DNA was positive in 13 cases, three cases were positive for HCV-RNA (indicating a healthy carrier rate of HCV-RNA of 4.1%), and seven cases were positive for HGV/GBV-C RNA. The incidence of these three virus-related markers was low in cases with fatty liver and micronodular cirrhosis, but in cases with chronic hepatitis, macronodular cirrhosis and hepatocellular carcinoma, the incidence of HBV-DNA and HCV-RNA increased with advancing disease. A positive rate of anti-HBs or anti-HBc (HBV-Ab) or both was found between 30 and 50% in all histopathological groups, and no noticeable relations between the positive rate and microscopical changes were detected. The presence of HGV/GBV-C RNA seemed to be unrelated to hepatic inflammation or generalized inflammatory changes or both occurring together. The decadal age incidence of the virus-related markers and their incidence in various hepatic diseases are also reported.

Liver weight of adult Japanese, especially recent weight values.

Cases of 445 adult Japanese autopsies of the Tokyo Metropolitan Medical Examiner's Office were used in this study. They were either negative for all hepatitis virus-related markers examined or had little or no histopathological hepatic changes. The maximum liver weight was observed in the fifth decade in both sexes, and after the fifth decade the liver weight decreased markedly with increasing age. The sexual difference in the liver weight was most predominant in the third to fifth decades, but the sexual difference was not marked in the older age groups. The highest liver weight to body weight ratio was observed in the fifth decade of both sexes, and a total decadal pattern of the ratio was similar to a parabola. An interesting finding was that the male liver weights in the third to fifth decades considerably increased in recent years, but the female liver weights in the third decade were almost the same despite the difference in investigation period. We suggest the data of this study may be a standard for Japanese people.

Apoptosis induced by microtubule disrupting drugs in cultured human lymphoma cells. Inhibitory effects of phorbol ester and zinc sulphate.

The effects of the microtubule disrupting drugs (MDD) vinblastine, vincristine and colchicine on a human lymphoma cell line, BM 13674, were investigated. Twelve hours after administration of vinblastine (10(-3) mg/ml), vincristine (10(-2) mg/ml) or colchicine (10(-2) mg/ml), cell death with the characteristic morphology of apoptosis was observed in 71.6%, 82.2% and 76.9% of the cells respectively. The mode of death was confirmed as apoptotic by the occurrence of internucleosomal DNA cleavage, which was demonstrated by agarose gel electrophoresis. For the purpose of casting light on the mechanism involved, inhibition tests were performed on apoptosis induced by one of these drugs, vinblastine, using a phorbol ester (PDBu), zinc sulphate and cycloheximide. PDBu, an activator of protein kinase C, and zinc sulphate, a putative inhibitor of the endonuclease were thought to be responsible for internucleosomal DNA cleavage; both markedly reduced the induction of apoptosis. The protein synthesis inhibitor cycloheximide, on the other hand, had no inhibitory effect. Moreover, cycloheximide treatment per se enhanced apoptosis. This suggests that new protein synthesis is not required for the execution of vinblastine-induced apoptosis. Such a finding is in accord with recent reports suggesting that the "death program" within many cell types may be primed but unable to proceed due to concomitant production of specific "apoptotic inhibitors". It is suggested that phorbol esters prevent vinblastine-induced apoptosis in the BM 13674 cells by activating one or more of these specific "apoptotic inhibitors", possibly by means of PKC-mediated phosphorylation.

An immunohistochemical and molecular biological study of c-erbB-2 amplification and prognostic relevance in gallbladder cancer.

Forty-three cases of gallbladder cancer were investigated for c-erbB-2 gene amplification and c-erbB-2 protein over-expression using a combined polymerase chain reaction (PCR) and immunohistochemical approach. Thirty out of 43 cases (69.6%) demonstrated c-erbB-2 gene amplification, the positive rates being 50% and 77.4% for twelve early cancers and thirty-one advanced cancers, respectively (P < 0.05). However, there was no statistically significant correlation between c-erbB-2 gene amplification and histologic grade of differentiation or lymph node metastasis. Fourteen out of 43 cases (32.6%) showed positive immunoreactivity reflecting c-erbB-2 protein over-expression but again no statistically significant correlation was found with grade of differentiation, invasion or lymph node metastasis. Neither the c-erbB-2 gene nor the protein revealed any close relation to prognosis. In contrast, histopathologic findings for histologic grade of differentiation, invasion grade and lymph node metastasis showed good correlations to prognosis and between themselves. In conclusion, while c-erbB-2 gene and protein in gallbladder cancers might be related to invasiveness, they are not applicable as predictive factors for prognosis.

Clonal Ig-gene rearrangement in some cases of gastric RLH detected by PCR method.

Clonal immunoglobulin (Ig) heavy chain gene rearrangement in gastric reactive lymphoid hyperplasia (RLH) cases was investigated by means of the 'double' polymerase chain reaction (PCR) using formalin-fixed and paraffin-embedded tissue. Rearranged DNA sequences, formed by combinations of variable (VH) and joining (JH) regions, were amplified with oligomeric primers. One microgram of DNA extracted from formalin-fixed and paraffin-embedded tissue was applied as the 'first PCR' template and one ten-thousandth of the first PCR product was used as the 'second PCR' template. As a control study for the double PCR method, DNA isolated from frank B cell gastric malignant lymphomas was assessed. Clear single bands between 100 and 150 base pair markers in length were evident on agarose gel electrophoresis in 10 out of 13 cases (76.9%) of malignant lymphomas while 2 out of 22 cases (9%) of RLHs revealed clear single bands of the same length, suggesting malignant lymphomas; however, no histologic features of malignant lymphomas were present. It is concluded that even gastric RLH cases satisfying histopathologic criteria for benign lymphoid hyperplasia may contain occult monoclonal B cell populations suggesting a continuous and progressive spectrum of lesions contributing to B cell neoplasia.

Effectiveness of a new drug, bredinin, on canine kidney allotransplant survival.

BR has remarkable effectiveness on canine kidney allograft survival. Its effectiveness in suppression of the immune responses is dependent on the dosage. Atrophy of the spleen and the mesenteric lymph nodes is prominent in treated dogs. Septic complications are slight, with no reduction in the number of peripheral white blood cells. Erosive changes of the mucosa in the entire bowel are the major side effects of the drug treatment. Liver functional abnormality has not been observed.

[Long-term toxicological studies with mizoribine (Bredinin) in beagle dogs (the second report). Pathological studies on visceral organs].

Four Beagle dogs were administrated with mizoribine 2.5 mg/kg/day for 36 months. Clinical and laboratory findings were fully described in the first report, in which no remarkable abnormalities were detected. Visceral organs of the experimental group and non-treated control group were submitted for pathological investigations. No conspicuous macroscopical and microscopical drug-induced changes were disclosed in the experimental group.

Is macronodular liver cirrhosis an irreversible lesion? Changes in the intrahepatic micro-vasculature treated with estradiol benzoate in rats.

Estradiol benzoate (EB), a synthetic estrogen, was injected to macronodular cirrhotic rats, and the reversibility of the lesion was investigated. Macronodular liver cirrhosis (MNLC) was produced by use of carbon tetrachloride plus 3'-Me-DAB. In animals with no treatment, regenerative nodules remained unchanged during an eighteen-week observation period. Microangiograms showed markedly deranged intrahepatic microvasculatures, including an irregularly contoured vascular tree and rarefaction in the peripheral area. On the other hand, EB treatment exerted conspicuous efficacy on the above-mentioned intrahepatic microcirculatory disorder and on sinusoidal dilatation, which led the livers with MNLC to revert to almost normal appearance. It is concluded that EB provides a salutary effect on the intrahepatic microcirculatory system, by means of which MNLC can be shown to be a reversible lesion.

Arterial infusion of combination therapy using dibutyryl adenosine 3',5'-monophosphate and mitomycin C for hepatocellular carcinoma occluding main portal vein: case studies.

Intraarterial infusion therapy combining dibutyryl adenosine 3',5'-monophosphate and mitomycin C was administered to 8 patients with hepatocellular carcinoma occluding the main portal vein. Three patients survived more than nine months, including 1 who has been well for forty-eight months with technical imaging showing complete regression. A decreased level of serum alpha-fetoprotein of more than 90% was obtained in 38% of patients. Causes of death included hepatic failure, rupture of esophageal varices, acute gastric ulcer, and cerebral hemorrhage. It is concluded that the combination therapy may be useful for HCC.

Chronic inhalation toxicity and carcinogenicity studies of 3-chloro-2-methylpropene in BDF1 mice.

Chronic toxicity and carcinogenicity studies of 3-chloro-2-methylpropene (CMP), which has been widely used as an insecticide and chemical intermediate, were carried out in BDF1 mice. CMP was administered to mice in groups of 50 male and 50 female mice by the inhalation route 5 days per week for 104 weeks at doses of 0, 50, 100 or 200 ppm. Male and female mice in the CMP-exposed groups had decreased body weight but no noticeable clinical signs when compared with the control group. Dose-related increases in the incidences of gastric mucosal hyperplasia and squamous cell papilloma were observed in both sexes, and squamous cell carcinoma was observed in only one male mouse in the 100 ppm group. An increased incidence of Harderian gland adenoma in female mice was also recognized. In the nasal cavity, eosinophilic exudate associated with atrophy of olfactory epithelia, respiratory metaplasia of olfactory epithelia and olfactory gland, and eosinophilic changes in respiratory and olfactory epithelia were increased in both sexes.

[The ultrastructural study on the relationship between Chlamydia trachomatis and the host cell in adhesion and during and after invasion].

Chlamydia trachomatis (C. trachomatis) is 0.4-0.5 microns in diameter in the EB stage, with the nucleoid located eccentrically within it. We studied the relationship between the eccentric nucleoid and the host cell surface under electron microscope, when C. trachomatis was in adhesion to the host cell and during and after invasion into the host cell. When C. trachomatis adheres to and invades a host cell, the eccentric nucleoid is located on the side distant from the host cell surface (44 of 66, 66%). The 1 x 3 chi-square test indicates that the difference is significant at p less than 0.001. After C. trachomatis invades a host cell, the eccentric nucleoid is located on the side close to the host cell surface (61 of 138, 44%). The 1 x 3 chi-square test shows the difference to be significant at p less than 0.001. However, since the latter (44%) is less than the former (66%). C. trachomatis revolves between adhesion and the completion of the invasion. The 2 x 3 chi-square test shows a significant difference at p less than 0.001.

[A case of Trichosporon beigelii peritonitis in CAPD].

Continuous ambulatory peritoneal dialysis (CAPD) was introduced to Japan ten years ago and was established as the treatment for end-stage renal disease along with HD. Although the incidence of peritonitis in CAPD has decreased by educating the patients and parents and the improvement of various devises of CAPD, peritonitis is still one of the major complications of CAPD. Fungus is a rare pathogen for peritonitis in CAPD, but it must be considered as a causative agent in cases of intractable peritonitis. This report describes the first case of Trichosporon beigelii (T. beigelii) peritonitis in CAPD in Japan. A nine year old boy with chronic renal failure due to bilateral vesicoureteral reflux was given CAPD treatment four years prior to admission. This patient had been admitted to our hospital frequently because of recurrent bacterial peritonitis. The peritonitis in CAPD was usually treated by changing the peritoneal fluid and antibiotic treatment. In this case T. beigelii was proved to be a pathogen of peritonitis by culture of CAPD fluid and also serum antibody titers. T. beigelii infection was successfully eradicated from the peritoneal cavity by administration of MCZ and by the removal of peritoneal catheter. The patient was switched from CAPD to HD. In the case of intractable peritonitis in CAPD, rare fungal pathogens such as T. beigelii must be considered as a causative agent.