Molecular role of ascorbate in enhancement of NO production in activated macrophage-like cell line, J774.1

Ascorbate-enhanced nitric oxide (NO) production in lipopolysaccharide (LPS)- and interferon-gamma (IFN-gamma)-activated macrophage J774.1 cells through the inducible nitric oxide synthase (iNOS) pathway. The iNOS gene was synergistically induced by LPS and IFN-gamma. The inductive mechanism of ascorbate on the iNOS gene was studied by examining the degradation of I kappa B alpha by Western blotting, activation of the nuclear factor kappa B (NF-kappa B) by gel shift assays, and protein levels of interferon regulatory factor 1 (IRF-1) in LPS- and IFN-gamma-activated cells. Ascorbate had no effect on the onset of either I kappa B alpha degradation or the nuclear translocation of NF-kappa B, but it delayed the recovery of I kappa B alpha. The prolonged degradation of I kappa B alpha caused by ascorbate in LPS- and IFN-gamma-activated cells paralleled elevated NF-kappa B binding to DNA, which led to an increase in the iNOS protein level. Ascorbate alone did not induce I kappa B alpha degradation or NF-kappa B activation. Furthermore, ascorbate exerted no effect on the expression of I kappa B alpha and ubiquitin genes in the activated cells. Ascorbate could modulate NF-kappa B DNA binding activity in response to combined LPS and IFN-gamma activation, which increases NO production in activated macrophages.     

Metabolism-dependent hepatotoxicity of methimazole in mice depleted of glutathione.

Methimazole (MMI) (>0.1 mmol kg(-1), p.o.) given in combination with DL-buthionine sulphoximine (BSO) (3 mmol kg(-1), i.p., 1 h before MMI administration), an inhibitor of glutathione (GSH) synthesis, caused liver injury in mice. The injury was characterized by centrilobular necrosis of hepatocytes and an increase in serum alanine transaminase (ALT) activity. Methionazole (2 mmol kg(-1)) alone resulted in only a marginal increase in serum ALT activity, but produced no histopathological changes in the liver. Pretreatment with hepatic cytochrome P-450 monooxygenase inhibitors--cobalt chloride, isosafrole, methoxsalen, metyrapone and piperonyl butoxide-prevented or tended to suppress the hepatotoxicity induced by MMI in combination with BSO. Treatment with N,N-dimethylaniline and ethyl methyl sulphide, competitive substrates of flavin-containing monooxygenases (FMO), also resulted in remarkable suppression of the hepatotoxicity caused by MMI in combination with BSO. These results suggest that MMI is activated by reactions mediated by both cytochrome P-450 monooxygenases and FMO, and that the inadequate rates of detoxification of the resulting metabolite are responsible for the hepatotoxicity in GSH-depleted mice.     

β,β′-Iminodipropionitrile toxicity in normal and congenitally neurofilament-deficient Japanese quails

Morphological effects of a neurotoxin, ,-iminodipropionitrile (IDPN) were analyzed in normal and cogenitally neurofilament (NF)-deficient Japanese quails. These quails (6 weeks old) were injected intraperitoneally with IDPN (0.2 g/kg body weight) three times every 3 days. They were necropsied at 10 to 12 days after the first injection. In normal quails, axonal swellings were observed histologically in the ventral motoneurons, ventral root, commissura grisea and spinal ganglion in the cervical and synsacral spinal cord. Electron microscopically, the changes consisted of increased NFs, with scattered mitochondria, smooth endoplasmic reticulum and microtubules. The myelin sheaths of the involved nerves were thinner than those of the normal axons. These lesions were similar to those induced by IDPN intoxication in mammalian experimental animals. In NF-deficient quails injected with IDPN, no axonal changes were detected. These findings suggested that IDPN selectively attacked the NFs.     

A case of urinary retention secondary to aseptic meningitis

A 46-year-old male was admitted to our hospital with headache, high fever and subsequent consciousness disturbance. Spinal fluid examination intimated aseptic meningitis. Not only these symptoms but bladder dysfunction was presented. The patient was treated with conservative therapy and bladder dysfunction was resolved a month and a half later. We report a case of urinary retention secondary to aseptic meningitis and review the clinical presentation and treatment.     

Significant antitumoral activity of cationic multilamellar liposomes containing human IFN-beta gene against human renal cell carcinoma.

PURPOSE: Immunotherapy is the most effective treatment against metastatic renal cell carcinoma (RCC). However, the response rate is approximately 15%. More effective therapy is, therefore, needed for patients with metastatic RCC. We then examined the antitumor effect of cationic multilamellar liposome containing human IFN-beta (huIFN-beta) gene (IAB-1) against RCC. EXPERIMENTAL DESIGN: Concentrations of huIFN-beta protein were measured by ELISA. The cytotoxicity of IAB-1 against human RCC (NC65, ACHN, and freshly isolated RCC cells), prostate and bladder cancer cell lines, and renal proximal tubule endothelial cells (RPTEC5899) was examined by the colorimetric method using tetrazolium salt. Apoptosis was assessed by the acridine-orange staining. For in vivo study, we used NC65 cells inoculated into severe combined immunodeficiency mouse. RESULTS: The RCC cells treated with IAB-1 secreted significant amounts of huIFN-beta protein continuously. Drastic in vitro cytotoxic effect of IAB-1 against RCC was observed. In contrast, treatment with 1000 IU/ml recombinant huIFN-beta protein resulted in weak cytotoxicity. The cytotoxic effect against prostate and bladder cancer cell lines was less than that against RCC. Furthermore, no significant cytotoxicity was observed in RPTEC5899 cells. Apoptosis was observed in the cells treated with IAB-1, but recombinant huIFN-beta failed to induce apoptosis. The size of NC65 tumors transfected with IAB-1 in mice was significantly smaller than that receiving injection of empty liposome or recombinant huIFN-beta protein. CONCLUSION: These findings indicate that IAB-1 may have an antitumor activity against human RCC by inducing apoptosis, suggesting its potential clinical application for gene therapy against RCC.     

Stage IV ovarian clear cell adenocarcinoma treated effectively by chemotherapy with etoposide and cisplatin (EP)

We report a case of stage IV ovarian clear cell adenocarcinoma (OCCA) in a 72-year-old woman who was treated postoperatively with etoposide combined with cisplatin (EP). The patient exhibited bulky intrapelvic and para-aortic lymph nodes with metastases to the cervical lymph nodes. The primary lesion was resected and, postoperatively, she received one course of combination chemotherapy consisting of cyclophosphamide, 500 mg/m²; doxorubicin, 50 mg/m²; and cisplatin, 70mg/m² (CAP), followed by six courses of combination chemotherapy consisting of etoposide, 80mg/m² days 1 through 5 and cisplatin, 70 mg/m² on day 5 every 4 weeks. After five courses of EP, the lymph node metastases had virtually disappeared. The patient is now disease-free 21 months after the initial surgery. These findings suggest that EP may be useful in treating OCCA.     

Primary transitional cell carcinoma of prostate: a case report

A 67-year-old male was admitted with a three-month history of voiding difficulty. Prostate specific antigen remained within the normal limit. Under the diagnosis of benign prostatic hypertrophy, transurethral resection of prostate was performed. Pathological examination of the resected specimens of the prostate revealed transitional cell carcinoma. Two courses of systemic M-VAC (methotrexate, vinblastine, doxorubicin, cisplatin) chemotherapy were performed, followed by cystoprostatourethrectomy, pelvic lymphadenectomy, and ileal conduit construction. Now one year has elapsed, with no clinical signs of recurrence.     

Stress and Coping Styles are Associated with Severe Fatigue in Medical Students

Fatigue is a common complaint among medical students and researchers consider it to be related to poor academic outcomes. The authors' goal in the present study was to determine whether stress and coping strategies were associated with fatigue in medical students. The study group consisted of 73 second-year healthy students attending the Osaka City University Graduate School of Medicine. Participants completed a questionnaire about fatigue (Japanese version of Chalder Fatigue Scale), stress, stress coping (Japanese version of the Coping Inventory for Stressful Situations), overwork, and nocturnal sleeping hours. On univariate and multivariate logistic regression analyses adjusted for age and gender, stress was positively associated with fatigue. In addition, after adjustment for age, gender, and emotion- and task-oriented stress coping activities, avoidance-oriented stress coping activity was associated with fatigue. The results suggest that stress and the coping style are correlated with fatigue in medical students.