Serum concentrations of resistin-like molecules beta and gamma are elevated in high-fat-fed and obese db/db mice, with increased production in the intestinal tract and bone marrow.

AIMS/HYPOTHESIS: Resistin and the resistin-like molecules (RELMs) comprise a novel class of cysteine-rich proteins. Among the RELMs, RELMbeta and RELMgamma are produced in non-adipocyte tissues, but the regulation of their expression and their physiological roles are largely unknown. We investigated in mice the tissue distribution and dimer formation of RELMbeta and RELMgamma and then examined whether their serum concentrations and tissue expression levels are related to insulin resistance. METHODS: Specific antibodies against RELMbeta and RELMgamma were generated. Dimer formation was examined using COS cells and the colon. RELMbeta and RELMgamma tissue localisation and expression levels were analysed by an RNase protection assay, immunoblotting and immunohistochemical study. Serum concentrations in high-fat-fed and db/db mice were also measured using the specific antibodies. RESULTS: The intestinal tract produces RELMbeta and RELMgamma, and colonic epithelial cells in particular express both RELMbeta and RELMgamma. In addition, RELMbeta and RELMgamma were shown to form a homodimer and a heterodimer with each other, in an overexpression system using cultured cells, and in mouse colon and serum. Serum RELMbeta and RELMgamma levels in high-fat-fed mice were markedly higher than those in mice fed normal chow. Serum RELMbeta and RELMgamma concentrations were also clearly higher in db/db mice than in lean littermates. Tissue expression levels revealed that elevated serum concentrations of RELMbeta and RELMgamma are attributable to increased production in the colon and bone marrow. CONCLUSIONS/INTERPRETATION: RELMbeta and RELMgamma form homo/heterodimers, which are secreted into the circulation. Serum concentrations of RELMbeta and RELMgamma may be a novel intestinal-tract-mediating regulator of insulin sensitivity, possibly involved in insulin resistance induced by obesity and a high-fat diet.

Dexamethasone stabilizes IGFBP-1 mRNA in primary cultures of rat hepatocytes.

We have shown that dexamethasone stimulates insulin-like growth factor binding protein-1 (IGFBP-1) production due to an increase in IGFBP-1 mRNA content in primary cultures of rat hepatocytes. In the present study, we investigated the effect of dexamethasone on the stabilization of IGFBP-1 mRNA in this system. We found that dexamethasone stabilized IGFBP-1 mRNA from the dot blot analysis with actinomycin D in the culture medium. It is suggested that the stabilization of IGFBP-1 mRNA is one of the mechanisms in which IGFBP-1 mRNA increases during dexamethasone treatment in primary cultures of rat hepatocytes.

Sexual differences in branched chain amino acid metabolism into fatty acids and cholesterol in Harderian gland of golden hamster.

The Harderian gland of golden hamster (Mesocricetus auratus) secretes copious lipids, most of which is 1-alkyl-2,3-diacylglycerol (ADG). We previously reported that the composition of ADG shows marked sexual dimorphism [Seyama et al. (1995) J. Biochem. 117, 661-670]. Male ADG contains only straight chain alkyl and acyl groups, but female ADG contains a lot of branched chain ones too. In this study, we investigated the metabolism of branched chain amino acids (BCAAs) and analyzed the incorporation of the metabolites into lipids in the Harderian gland. Golden hamsters were injected intraperitoneally with [U-14C]BCAAs, and Harderian glands were obtained at 3, 6, 9, and 24 h after injection. Lipids were then extracted from the glands and analyzed. Thin layer chromatography revealed that the ADG was labeled in both sexes, but the profile depended on the sex. The cholesterol fraction was labeled only in the male gland. The alkyl and acyl groups of ADG were subjected to radio-gas liquid chromatography. As for the alkyl groups, radioactivity was detected in straight-C16 and -C18 chains in males, while branched-C17 and -C19 chains were labeled in females. As for the acyl groups, straight-C14, -C15, and -C16 chains were labeled in males, while in females, branched-C17 and -C19 chains were labeled as well as a straight-C16 chain. These results suggest that the BCAA metabolism should be regulated as to the sex at the step of branched chain acyl-CoA degradation in the Harderian gland of golden hamster, which causes the sexual dimorphism in the lipid composition in this gland.

Interaction between cAMP-dependent and insulin-dependent signal pathways in tyrosine phosphorylation in primary cultures of rat hepatocytes.

The present studies were undertaken to determine whether the interaction between cAMP-dependent and insulin-dependent pathways in primary cultures of rat hepatocytes affects biological functions and tyrosine phosphorylation. Quiescent hepatocytes were pretreated with dibutyryl cAMP or cAMP-generating agents such as glucagon, and then treated or not with insulin. Preincubation for 6 h with dibutyryl cAMP or glucagon enhanced the effect of insulin on DNA synthesis, but not the effect of insulin on amino acid transport or glycogen and protein synthesis. Tyrosine phosphorylation of intracellular proteins was determined by immunoblot analysis using an anti-phosphotyrosine antibody. Maximum tyrosine phosphorylation of a 195 kDa protein, which may be a substrate of insulin receptor kinase, of 175-180 kDa proteins, including insulin receptor substrate (IRS)-1, and of 90-95 kDa proteins, including the insulin receptor beta-subunit, was reached within 30 s of incubation with insulin. Pretreatment for about 3 h with dibutyryl cAMP or cAMP-generating agents clearly increased insulin-dependent tyrosine phosphorylation of the 195 kDa protein, but not IRS-1, IRS-2 or the insulin receptor beta-subunit. Because dibutyryl cAMP and cAMP-generating agents did not increase insulin receptor number or its kinase activity, the effect of cAMP on this potentiation of tyrosine phosphorylation is assumed to be exerted at a step distal to insulin receptor kinase activation. The potentiation by cAMP pretreatment of insulin-stimulated tyrosine phosphorylation may in part be secondary to inhibition of phosphotyrosine phosphatase activity, because cAMP pretreatment blunted the effect of Na3VO4 on the net tyrosine phosphorylation of the 195 kDa protein as compared with cells pretreated with no additive. In summary, the interactions between cAMP-dependent and insulin-dependent pathways that lead to augmentation of DNA synthesis appear to parallel the changes in tyrosine phosphorylation. Further studies will be required to determine whether there is a causal relationship between these phenomena.

Production of insulin-like growth factors and their binding proteins in primary cultures of rat liver parenchymal and nonparenchymal cells.

Regulation of the production of insulin-like growth factor (IGF)-I, IGF-II, IGF binding proteins (IGFBPs), and their related proteins by various hormones was investigated in primary cultures of rat liver parenchymal and nonparenchymal cells. Freshly isolated parenchymal cells contained mRNAs of IGF-I, IGF-II, IGFBP-1, IGFBP-4, growth hormone (GH) receptor, and the acid-labile subunit (ALS), which forms a ternary complex with IGF-I and IGFBP-3; however, parenchymal cells did not express the IGFBP-3 gene. In contrast, nonparenchymal cells contained IGFBP-3 mRNA exclusively, as we reported previously [Takenaka et al. Agric. Biol. Chem., 55, 1191-1193 (1991)]. Cultured rat parenchymal cells produced IGF-I, IGFBP-1, and IGFBP-4 prominently. In these cells, secretion of IGF-I and the content of IGF-I mRNA was greatly increased in the presence of GH in the medium. Insulin also increased the production of IGF-I. Secretion of IGFBP-1 into the medium was enhanced by treatment with glucagon, dibutyrylcyclic AMP (Bu2cAMP), and dexamethasone (Dex) and these enhancements with glucagon and Dex reflected the increase in its mRNA content. Insulin depressed the secretion of IGFBP-1. The content of IGFBP-4 in the parenchymal cells was increased by insulin, Bu2cAMP, and triiodothyronine (T3), thereby enhancing the production of IGFBP-4 and secretion into the medium. Cultured liver nonparenchymal cells of rats produced IGFBP-1, IGFBP-3, and IGFBP-4. Secretion of IGFBP-1 was increased by Bu2cAMP in the medium, that of IGFBP-3 by IGF-I, and that of IGFBP-4 by both IGF-I and Bu2cAMP. Regulation of the production of IGFBP-3 by IGF-I was demonstrated in these investigations. These results suggest that GH increases production of IGF-I in the parenchymal cells and this IGF-I, in turn, increases the production of IGFBP-3 in nonparenchymal cells. As we found GH also increases ALS production in parenchymal cells, by these mechanisms, GH increases the formation of the ternary complex of IGF-I, IGFBP-3, and ALS. This study clearly demonstrates the interrelationship between parenchymal and nonparenchymal cells in the production of IGF-I and IGFBPs in the liver.

[Extracorporeal shock wave lithotripsy monotherapy using Lithostar--factors influencing actuarial residual stone rates].

For the evaluation of post treatment course after initial extracorporeal shock wave lithotripsy (ESWL) as a monotherapy using the Lithostar, the actuarial residual stone rate (ARSR) was calculated by the Kaplan-Meier formula in 713 calculi (255 renal and 458 ureteral calculi). ARSRs in all calculi at 3, 6, 12 and 48 months were 41.6, 30.5, 20.6 and 10.3%, respectively. Significantly higher ARSRs were observed in the following former group compared to the latter: large calculi to small or middle sized calculi, renal calculi to ureteral calculi (small or middle sized), and plural renal calculi (middle sized) to single renal calculi (middle sized). There was no significant difference either between large renal and large ureteral calculi, or overall single and plural calculi. These findings show that ARSRs calculated by the Kaplan-Meier formula clearly reveal factors influencing the ESWL effects.

[Extracorporeal shock wave lithotripsy on patients with spinal cord injury with special reference to autonomic hyperreflexia].

We performed treatment of urolithiasis on 7 patients with spinal cord injury (6 males and one female, with a mean age of 41 years old) by extracorporeal shock wave lithotripsy (ESWL) using a Lithostar (Siemens). The level of injury was cervical in 3 patients and lower thoracic in 4. The treated stones were renal in 6 patients, including one staghorn caliculus, ureteral in 2, and bladder stones in 2 patients. ESWL was performed under general anesthesia in 2 patients, and in another patient, epidural anaesthesia was employed in the first several sessions, but thereafter treatment was safely continued with no anesthesia. The other 4 patients were treated without anesthesia. In most patients, the stones (9 stones in 6 cases) were easily disintegrated after treatment of from one to 14 sessions, except one case of staghorn caliculus, and the clearance of fragments was also satisfactory. In one patient, hypertension and bradycardia due to autonomic hyperreflexia were observed during ESWL, and the treatment was discontinued, but the stone was successfully disintegrated and the fragments were voided. In another patient, autonomic hyperreflexia was observed while the fragments passed through the ureter, although no signs of this reflexia were seen during the ESWL procedure.

[Dynamic study of the hormonal levels and tumor markers after the first administration of long-acting LH-RH analogue in patients with prostate cancer].

The dynamics of hormonal levels and tumor markers after the first administration of long-acting luteinizing hormone-releasing hormone (LH-RH) analogue were evaluated in patients with prostate cancer. Eight patients with histopathologically proved prostate cancer who were previously untreated were studied. The surge in plasma testosterone was recognized in 7 patients after the first administration of a long-acting LH-RH analogue, and reached the highest level after the 3rd day in 6 patients and 14th day in 1 patient. The onset of flare-up reaction due to a transient increase in plasma testosterone was recognized in 3 patients, whose clinical symptoms and signs were increased bone pain in 2 patients and acute urinary retention in 1 patient. An abnormal level of serum prostatic acid phosphatase (PAP) and prostate specific antigen (PSA) was observed in 7 of the 8 patients before treatment. The serum PAP and PSA levels slightly increased after treatment in 4 and 3 patients, respectively. These findings suggest that the combination of estrogen or antiandrogen would allow a safer use of long-acting LH-RH analogue to prevent the risk of a flare-up reaction associated with the first administration of long-acting LH-RH analogue.

[Epidemiological study on sexually transmitted diseases in Saitama Prefecture--mainly study of male urethritis].

Epidemiological studies on sexually transmitted diseases (STDs) in Saitama Prefecture, from 1989 through 1991, were performed and the following results were obtained. The number of patients was 1,833 (male: 1,532, female: 30) cases in 1989, 1,294 (male: 1,163, female: 131) cases in 1990, and 1,672 (male: 1,478, female: 149) cases in 1991. Male patients in their twenties or thirties were often affected by STDs. The order of frequency of STDs was roughly as follows; non-gonococcal and non-chlamydial urethritis, gonococcal urethritis, chlamydial rethritis, condyloma acuminatum, genital herpeic infection and syphilis. Concerning the criteria of the diagnosis on chlamydial urethritis, cases diagnosed by only clinical findings decreased markedly from 1989 through 1991, cases diagnosed by the combination of the antibody of C. trachomatis and clinical findings increased, and cases diagnosed by the detection of C. trachomatis were always about 70%. Male patients were mostly infected from prostitutes. Many patients with STDs were often infected in Taiwan, the Philippines, the Kingdom of Thailand and Republic of Korea.

Myofibroblastic pseudotumor mimicking epididymal sarcoma. A clinicopathologic study of three cases.

The clinical and pathologic features of three cases of epididymal pseudotumor are described with emphasis on its differential diagnosis and histopathogenesis. The first lesion was found histologically worrisome in the region of ductus epididymis by a torsioned testis in an 8-month-old boy, although the lesion was clinically indistinctive, and grossly not remarked. Histodiagnostic difficulties are encountered because this lesion forms numerous infiltrative spindle cells in haphazard arrangement. Ultrastructurally, the cell components of this lesion are predominantly myofibroblasts, intermediate cells between fibroblasts and myocytes. Similar epididymal lesions with the same cell components were identified in the two other infantile cases on the review of 36 consecutive orchiectomy specimens all obtained for testicular torsion. These three boys were found to present with a relatively gradual clinical course of the testicular torsion. It is suggested that myofibroblastic proliferation in the epididymal location histologically reminiscent of sarcomas occurs under ischemic circumstances.

[A report of two cases of multilocular cystic renal cell carcinoma: review of 51 cases reported and the results of a prognostic survey].

We report 2 cases of multilocular cystic renal cell carcinoma. One was in a 33-year-old male, presenting with ultrasonic abnormality of the left kidney at an annual employee health care examination. Computerized tomography (CT) demonstrated a 5 cm of multilocular cystic mass adjacent to the lower pole of the left kidney. Another was in a 44-year-old male, presenting with microscopic hematuria at an annual employee health care examination. CT of the abdomen revealed a 6.5 cm of multilocular cystic mass on the upper pole of the right kidney. Both were diagnosed as renal cell carcinoma by the angiography and underwent radical nephrectomy. Gross specimens showed typical multilocular cystic appearance and histopathology showed clear cell carcinoma infiltrating septa and replacing epithelium of the cyst walls. Both patients are alive without evidence of disease at, 21 months and 14 months after operation, respectively. Including our cases, 51 multilocular cystic renal cell carcinoma and multilocular cystic nephroma associated with renal cell carcinoma have been reported. From the review of the literatures and the answer of the questionnaires inquiring about the outcome of the patient to Japanese reporters, the outcome of 38 patients was ascertained. The 10-year survival rates and non-recurrence rate after operation calculated by the Kaphan-Meier formula were 97.3% and 90.3%, respectively. Because of the good prognosis of reported cases, we concluded that we should choose kidney-sparing surgery for the operation of multilocular cystic renal cell carcinoma.

[Efficacy of sodium imipenem/cilastatin on patients of complicated urinary tract infections--following the failure of prior antimicrobial agents].

Clinical efficacy of sodium imipenem/cilastatin was studied on 45 patients with complicated urinary tract infections, on whom prior antimicrobial agents were ineffective. A 0.5g dose of IPM/CS was administered by drop infusion twice a day for 5 days. The clinical efficacy was evaluated according to the criteria of the Japanese UTI committee. Of a total of 65 strains, 29 strains of gram positive bacteria and 36 strains of gram negative bacteria, were detected as causative microbials. Bacteriologically, 23 strains of gram positive bacteria (79%) and 34 strains of gram negative bacteria (94%) were eradicated following the treatment. S. aureus, E. faecalis and Flavobacterium sp. were less sensitive to IPM/CS. Overall clinical effectiveness rate of IPM/CS in the present study was 87%. Adverse drug side effects were observed in six patients, namely they were eruption, headache and slight elevation of serum transaminase. These findings suggest that IPM/CS is an effective agent for the treatment of complicated UTI even when the prior medicine was ineffective.

[A case of emphysematous pyelonephritis with disseminated intravascular coagulation].

A case of emphysematous pyelonephritis with disseminated intravascular coagulation (DIC) is presented. A 54-year-old woman was admitted to our hospital because of unclear consciousness and extremely high blood glucose level. The laboratory data suggested uncontrolled diabetes mellitus (DM) and urinary tract infection with sepsis and DIC. The plain abdominal X-P and abdominal CT revealed the existence of gas in the right renal parenchyma, perinephric tissue and the upper part of the right ureter. Right nephrectomy was performed after the improvement of the patient's condition by the echo-guided drainage of the right kidney and the treatment for infection, DM and DIC. We reviewed 71 cases of emphysematous pyelonephritis in the Japanese literature and the choice of treatment was discussed.

[Significance of prostatic acid phosphatase, gamma-seminoprotein and prostatic specific antigen in the urine. First report: the measurement of PAP, gamma-Sm and PA in the urine of patients with prostatic diseases].

To study the significance of prostatic acid phosphatase (PAP), gamma-seminoprotein (gamma-Sm) and prostatic specific antigen (PA) in urine, we have determined the urinary levels of these proteins in women and infants, in patients without prostatic disease, in patients with benign prostatic hypertrophy, and in patients with prostatic adenocarcinoma. Women and infants were found to excrete little PAP (27.9 +/- 4.8 ng/mg) and undetectable levels of gamma-Sm except one case, and undetectable levels of PA in the urine. The excretion of PAP in patients with prostatic carcinoma who were either castrated, or treated with endocrine therapy was lower than the levels in women and infants, or the levels in patients without prostatic diseases, or the levels in patients with BPH. Urinary excretion levels of gamma-Sm and PA were undetectable in the patients with well-controlled prostatic carcinoma. The present study suggests that the determination of PAP, gamma-Sm and PA in the urine of patients with prostatic carcinoma may become a useful tool for monitoring of the primary locus of the carcinoma, but additional assays of urinary PAP, gamma-Sm and PA should be measured at regular intervals to be concluded.

Purification and properties of gamma-glutamyl transpeptidase from the tissue of human benign prostatic hypertrophy.

The enzyme of gamma-glutamyl transpeptidase was purified to homogeneity from the tissue of human benign prostatic hypertrophy and its enzyme properties were studied. The enzyme activity was detected mainly in the luminal border of the epithelium lining ducts by histochemical staining. The enzyme was purified 759-fold that of the crude extract. The specific activity of the purified enzyme was 79,900 mU/mg protein. The following enzyme properties were obtained: Michaelis constant of the enzyme was 0.83 mmol/l. The molecular weight was 72 kDa, consisting of two subunits, 45 kDa and 27 kDa. The isoelectric point of the enzyme was 8.5. The optimum pH ranged from 8.2 to 8.5. By Concanavalin A-sepharose affinity chromatography, more than 60% of the enzyme activity was eluted in the weakly bound fraction, suggesting biantennary complex sugar chain was the major type among the asparagine-linked sugar-chains of the enzyme.

Possible metastatic routes via portacaval shunts in renal adenocarcinoma with liver metastasis.

Renal adenocarcinoma findings from autopsies on patients with and without liver metastasis (635 and 936 patients, respectively) were investigated concerning the mode of metastasis. The patients with liver metastasis showed a significantly higher frequency of metastases to the lungs, lymph nodes, contralateral kidney, adrenals, pancreas, spleen, peritoneum, and intestines; in the female patients, those with liver metastasis had a higher frequency of metastasis to the ovary and uterus than patients without liver metastasis. Some of the metastases to those organs are explainable by venous spread via portacaval shunts as well as ordinary hematogenous or lymphogenous spread, especially, those to the contralateral kidney, adrenals, spleen, intestines, or ovary.

[A renal cell carcinoma mimicking a multilocular renal cyst].

A 38-year old man was admitted to the hospital for an investigation of abnormal ultrasonographic findings of the left kidney. Ultrasonography demonstrated multiple, fluid-filled masses separated by a highly echogenic septa and CT revealed a large cystic mass and a thick, irregular wall in the upper and middle poles. Selective, left renal arteriography showed a hypovascular mass. Thus, a presumptive diagnosis was made of multilocular renal cysts. Taking into accounted the possibility of a renal cell carcinoma. Surgical exploration and a left nephrectomy was performed. Microscopically, a multiloculated renal cell carcinoma thus was diagnosed, A definite, preoperative diagnosis is very difficulty to make in most cases, and surgical exploration is considered to be the only way to achieve an accurate diagnosis. Treatment for a multiloculated cystic renal mass containing multilocular renal cysts is a total or partial nephrectomy.

Prostate mucinous adenocarcinoma with signet ring cell.

Carcinoma exhibiting signet ring cell appearance in the prostatic gland is rare. An example of this rare tumor is presented herein.

Two different lymph node metastatic patterns of a prostatic cancer.

Among 753 autopsy prostatic cancer cases with a metastasis, 476 (63%) had a lymph node metastasis, whereas 277 (37%) did not. Two different lymph node metastatic patterns were observed: Type 1, combined metastasis involving the pelvic and paraaortic lymph nodes; and Type 2, metastasis to the paraaortic lymph nodes, but not to the pelvic lymph nodes. Type 1 metastasis cases showed a significantly more frequent metastasis to the bladder and rectum, and a less frequent metastasis to the lungs and liver. Hydronephrosis occurred more frequently (P less than 0.01) in the Type 1. Furthermore, in the Type 1 cases the lymph node metastasis appeared to be continuously invasive, but in the Type 2 cases, metastasis appeared to be the skip type or some metastases may have spread via the vertebral vein bypass route and may have been associated with a hematogenous metastasis.