Detection of beta-lactamase-producing strains isolated from urinary tract and their drug susceptibility

A total of 518 bacterial strains isolated from the urine of patients with various urological diseases in our Urological Department between November, 1987 and February, 1989 were studied for their beta-lactamase production and susceptibility to various antimicrobial agents was determined. beta-lactamase activity was determined by the acidometry disc method. There were 241 gram-positive cocci, 276 of gram-negative rods and 1 Neisseria gonorrhoeae. Thirty-four percent of the gram positive and 76.3% of gram negative rods produced beta-lactamase. S. aureus (81.3%), S. epidermidis (65.1%) in gram-positive cocci, E. cloacae (100%), S. marcescens (100%), C. freundii (100%), P. aeruginosa (97.2%), P. Rettgeri (88.9%), E. gergoviae (85.7%), K. oxytoca (84.6%), M. morganii (81.8%) and E. coli (69.0%) in gram-negative rods produced beta-lactamase at a higher rate. beta-lactamase produced by gram-positive cocci was entirely penicillinase, and that produced by gram-negative rods only penicillinase in 4.0%, only cephalosporinase in 44.2% and both in 25.4%. In S. aureus and S. epidermidis, the isolated rate of strains resistant to ampicillin (p less than 0.01) and piperacillin (p less than 0.05) in the beta-lactamase producing strains was significantly higher than that in the beta-lactamase non-producing strains. In E. coli, the isolation rate of strains resistant to ampicillin and piperacillin in the penicillinase-producing strains was significantly higher than in the penicillinase non-producing strains (p less than 0.01). But both cephalosporinase-producing strains and beta-lactamase non-producing strains showed high susceptibility to cephalothin. These results suggest that the penicillinase might present a clinical problem in the treatment of urinary tract infections by S. aureus, S. epidermidis and E. coli.     

Internalization of Mycobacterium bovis Bacillus Calmette-Guérin into Osteoblast-Like MC3T3-E1 Cells and Bone Resorptive Responses of the Cells against the Infection

Mycobacterium bovis BCG (BCG) is a live vaccine used worldwide against tuberculosis. However, it has unfavourable side effects such as osteitis or osteomyelitis, and these sometimes lead to vertebral caries in some patients as a result of bone resorption. Osteoblasts might play a role in the bone resorption caused by BCG infection, because they are central cells in bone metabolism. Cultured osteoblast-like cell lines (MC3T3-E1) derived from C57BL mice susceptible to BCG infection cells were infected with BCG at several doses. Interestingly, internalization of BCG-enveloped phagosome-like membrane in osteoblast-like cells were observed by transmission electron microscopy (TEM). Owing to infection, the proliferation and alkaline phosphatase activity of the osteoblast-like cells were reduced in a dose-dependent manner. On the other hand, interleukin (IL)-6 production was considerably enhanced by infection. These results suggest that BCG infects osteoblasts, suppressing their proliferation and differentiation and inducing bone resorption, which may be related to osteitis/osteomyelitis and bone caries caused by BCG infection.     

Brain Metastasis from Prostate Cancer: A Case Report

Brain metastases from prostate cancer are rare in postmortem examinations, and even rarer in clinical series. We report an unusual case of brain metastasis from prostate cancer confirmed by antemortem diagnosis in a 72-year-old man. The metastatic brain tumor was surgically resected and the patient was kept stable for more than 19 months after diagnosis of the brain metastasis.     

Measurement of proliferating cell nuclear antigen (PCNA) and its clinical application in oral cancers.

The PCNA score was measured in oral squamous cell carcinoma (SCC), and its relationship to other cell proliferation markers, Ki-67 score, S-phase fraction (SPF), and AgNORs counts was investigated. The PCNA score ranged from 0.4% to 43.5% with an average value of 22.8%, the Ki-67 score ranged from 4.9% to 40% with an average of 24.1%, and the SPF ranged from 0.4% to 32.5% with an average of 12.4%, while AgNORs counts ranged from 2.53/nucleus to 7.03/nucleus with an average of 4.74/nucleus. These four parameters were closely interrelated. There was a significant difference in PCNA score between malignant and nonmalignant lesions, suggesting a difference in growth activity. The mean PCNA score decreased significantly from 20.0% to 8.0% after cancer chemotherapy. The response of cancer cells to anticancer agents may be estimated by consecutive measurement of PCNA, since the PCNA score dropped after treatment in cases showing a favorable prognosis.     

Effects of ethanol feeding on collagen synthesizing and degrading enzymes in rat pancreas

Collagen metabolism in the pancreas was investigated in male Wistar strain rats after 7 weeks of ethanol feeding. Compared with control rats, the ethanol-fed rats had a normal hydroxyproline content in the pancreas. However, prolyl hydroxylase activity and collagenolytic cathepsin activity were increased, though collagenase activity did not change. Both prolyl hydroxylase activity and collagenolytic cathepsin activity were inversely correlated with amylase activities. These findings were also confirmed in ethanol-pyrazole treated rats. These results suggest that the ethanol-induced pancreatic injury, even at an early stage, accelerates the collagen metabolism in the pancreas.     

Menaquinone (vitamin K2)-6 production by mutants of Flavobacterium meningosepticum

Flavobacterium meningosepticum IFO 12535, a menaquinone (MK) producer, was mutagenized to improve productivity. A mutant, which was resistant to 1-hydroxy-2-naphthoate (HNA), was found to produce MK more abundantly: 34 mg/liter of culture broth and 5.5 mg/g of dry cells. The mutant was less sensitive to inhibition by HNA on MK biosynthesis than the wild-type strain. MK was isolated from cells of the mutant and identified as MK-6 based on its physicochemical characteristics. Mutants, which were given each KCN resistance, aromatic amino acid auxotrophy and no carotenoid productivity, did not show further increase of productivity.     

A 6-day subrenal capsule assay for predictive testing of primary esophageal cancers

Subrenal Capsule Assay (SRCA) as a chemosensitivity test was performed on 14 esophageal squamous cell carcinomas in order to select a more effective form of chemotherapy. Of the 14 assays, 12 were evaluable. Mice were treated with anticancer agents (e.g. Cisplatin, Bleomycin, Methotrexate, Vindesine) on days 1 and 3 after transplantation, and on day 6, the sensitivities were determined. Fresh esophageal cancers yielded an evaluable assay rate of 74%. The implant grew progressively for six days in the remaining group of control mice. Histologically, host cell infiltration at the border of the implant was observed from day 3 after transplantation, and cells had degenerated or had been partially replaced by scar tissue by day 6. The results of chemosensitivity tests differed according to the anticancer agent used or from case to case. Clinically, correspondence between the assay results and clinical results was obtained in 5 out of 7 cases. SRCA is a new promising chemosensitivity test which is clinically useful, and the present results indicated the feasibility of its use in developing an effective chemotherapy for esophageal cancer.     

The localization of CD44 and moesin in osteoclasts after calcitonin administration in mouse tibiae

We investigated the immunohistochemical localization of CD44, hyaluronate receptor, and moesin, of the ezrinradixin-moesin (ERM) family, in osteoclasts after calcitonin adminstration using confocal laser scanning microscopy and transmission electron microscopy to clarify the role of CD44 and moesin in their cytoskeletal organization and cell polarity. We also elucidated the localization of osteopontin (OPN) to confirm its possible role in cell-matrix recognition via CD44. In untreated mice, intense immunoreactivities for CD44 and moesin were detected on the basolateral plasma membrane of osteoclasts. Rhodamine-phalloidin reactivity was seen in a bandlike pattern on the region of contact between osteoclasts and bone and was also detected moderately along their basolateral plasma membrane. At 30 min after calcitonin administration, osteoclasts did not show either clear zones or ruffled borders. The bandlike reactivity of rhodamine-phalloidin in the contact region was diminished, although labeling was seen along osteoclasts. CD44 and moesin were colocalized along their plasma membranes, including the region facing the bone surface. Electron microscopic observation revealed that the microvillus processes in the contacting region with bone surface, as well as the basolateral plasma membrane, showed immunoreactivities to CD44 and moesin. At 60 min, some osteoclasts attached to bone and showed a bandlike pattern of rhodamine-phalloidin. On the other hand, OPN was localized under CD44-positive cytoplasmic processes and the clear zone of osteoclasts. These findings suggest that calcitonin effects on the cell polarity of osteoclasts and the CD44-moesin-actin filament system in osteoclasts plays an imporant role in cell polarity and cell-matrix recognition.