Comparative proteome analysis of two antagonist Bacillus subtilis strains.

Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB- 1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-1, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

Clinical epidemiology of ciprofloxacin-resistant Proteus mirabilis isolated from urine samples of hospitalised patients.

This study investigated the clinical characteristics of ciprofloxacin-resistant Proteus mirabilis isolates from urine samples associated with nosocomial infection or colonisation, and identified the risk-factors for ciprofloxacin resistance. Data for patients with ciprofloxacin-resistant P. mirabilis isolates (n=13) were compared with those for randomly selected patients with ciprofloxacin-susceptible P. mirabilis isolates (n=40) who were matched by temporal occurrence as control patients. The majority of ciprofloxacin-resistant P. mirabilis isolates were multiresistant, and ciprofloxacin resistance was associated significantly with previous use of fluoroquinolones and production of extended-spectrum beta-lactamases.

cRGD peptide-installed epirubicin-loaded polymeric micelles for effective targeted therapy against brain tumors.

Current therapeutic strategies against glioblastoma multiforme (GBM) are futile mainly because of the poor access of drugs into malignant tissues, which is hindered by the tight blood-brain tumor barrier in the GBM vasculature. Nanomedicines have shown potential for circumventing the vascular barriers of GBM, particularly by targeting markers on the luminal side of endothelial cells in the blood vessels of GBM for achieving effective and selective translocation into the tumor. Thus, as the αvß3 and αvß5 integrins overexpressed on the endothelial cells of GBM can be targeted by cyclic-Arg-Gly-Asp (cRGD) peptide, herein, we developed cRGD-installed micellar nanomedicines loading epirubicin, the potent antiglioblastoma agent, through a pH-sensitive hydrazone-bond for effective treatment of GBM. These cRGD-installed epirubicin-loaded polymeric micelles (cRGD-Epi/m) achieved faster and higher penetration into U87MG cell-derived 3D-spheroids than the micelles without cRGD, conceivably through a cRGD-integrin mediated pathway. In vivo, the cRGD-installed micelles effectively suppressed the growth of an orthotopic GBM model by delivering high levels of epirubicin throughout the tumor tissue. These results indicate significant prospects for cRGD-Epi/m as an effective and translationable treatment against GBM.

Interference of Long-Distance Movement of Grapevine berry inner necrosis virus in Transgenic Plants Expressing a Defective Movement Protein of Apple chlorotic leaf spot virus.

ABSTRACT Transgenic Nicotiana occidentalis plants expressing a movement protein (P50) and partially functional deletion mutants (DeltaA and DeltaC) of the Apple chlorotic leaf spot virus (ACLSV) showed resistance to Grapevine berry inner necrosis virus (GINV). The resistance is highly effective and GINV was below the level of detection in both inoculated and uninoculated upper leaves. In contrast, GINV accumulated in inoculated and uninoculated leaves of nontransgenic (NT) plants and transgenic plants expressing a dysfunctional mutant (DeltaG). On the other hand, in some plants of a transgenic plant line expressing a deletion mutant (DeltaA', deletion of the C-terminal 42 amino acids), GINV could spread in inoculated leaves, but not move into uninoculated leaves. In a tissue blot hybridization analysis of DeltaA'-plants inoculated with GINV, virus could be detected in leaf blade, midribs, and petiole of inoculated leaves, but neither in stems immediately above inoculated leaves nor in any tissues of uninoculated leaves. Immunohistochemical analysis of GINV-inoculated leaves of DeltaA'-plants showed that GINV could invade into phloem parenchyma cells through bundle sheath of minor veins, suggesting that the long-distance transport of GINV might be inhibited between the phloem cells and sieve element (and/or within sieve element) rather than bundle sheath-phloem interfaces. Immunogold electron microscopy using an anti-P50 antiserum showed that P50 accumulated on the parietal layer of sieve elements and on sieve plates. The results suggested that resistance in P50-transgenic plants to GINV is due to the interference of both long-distance and cell-to-cell movement of the virus.

Enhancement of the specific growth rate of Thiobacillus ferrooxidans by diatomaceous earth.

Diatomaceous earth is an effective carrier with a large surface area on which Thiobacillus ferrooxidans adsorbs, and enhances the oxidation rate of ferrous ions by the bacterium. The adsorption of T. ferrooxidans on diatomaceous earth was expressed by the Henry isotherm, and reached equilibrium within five minutes. The enhancement was due to an increase in the growth rate of the adsorbed cells, which were so active that their specific growth rate calculated using a Monod-type equation was higher than that of the free cells in the liquid phase.

Recurrent death among early gastric cancer patients: 20-years' experience.

Six hundred and ninety-six patients, from among a total of 1,868 cases, were diagnosed as having gastric cancer, and underwent surgical treatment at this Medical Institution, between 1972 and 1992. Eighteen patients (2.6%) died of recurrent disease, four (0.6%) tumors being mucosal, and fourteen (2.0%) submucosal, in accordance with depth of invasion. The clinico-pathological findings were analysed to clarify the risk factors of recurrence in patients with early gastric cancer, as follows: (a) submucosal invasion, (b) mixed type IIa+IIc in gross appearance, (c) tumor size more than 40 mm in diameter. In mucosal cancers, recurrence occurred in patients with lymph-vessel invasion and lymph-node metastasis. In terms of the site of the recurrent lesion, the patients had predominantly hepatic recurrence. We conclude that there are specific factors related to the increased risk of recurrence, that can be evaluated pre-operatively. By identifying the risk factors of recurrence, we can optimize the therapeutic approach to these patients.

Occurrence of toxicity and cell proliferation after a single gavage administration of chloroform to male F344 rats.

Chloroform, an industrial solvent and one of the most common environmental contaminants which produces carcinogenic effects in the liver and kidney of rodents, is not genotoxic in most traditional bacterial and mammalian test systems. Its carcinogenic potential appears attributable to the sustained cell turnover (regenerative hyperplasia) which results from chronic chloroform toxicity. In this present study, cell proliferation (replicative DNA synthesis, RDS) and histopathological changes in hepatocytes and renal tubular epithelial cells were assessed in male F344 rats following a single gavage chloroform exposure (50, 150 or 500 mg/kg). In addition, biochemical parameters (BUN, GOT, LDH and NAG) were examined using plasma and urine samples. Cell proliferation and histopathological changes (e.g. hypertrophy, necrosis, vacuolation) were only seen at the dose of 500 mg/kg in the liver and kidney. At the same dose, all biochemical markers were increased at the 24 to 48 hr time points. These results obtained are thus in line with earlier findings pointing to epigenetic carcinogenicity.

Mucinous cystic tumor of the retroperitoneum. A report of two cases.

Two cases of retroperitoneal mucinous cystic tumors and some diagnostic problems on aspiration cytology are presented. The first was a mixed benign, borderline and malignant mucinous tumor, and the second was a mixed benign and borderline mucinous tumor. Our knowledge of the cytology of ovarian mucinous cystic tumors was not helpful in making a correct diagnosis in these cases. Based on our experience, we emphasize the following. First, consider methods of collecting enough cells to make a diagnosis. Second, consider the possibility of mixed histologic features. Direct aspiration, if possible, from papillary lesions inside the cyst will probably lead to an accurate diagnosis. Measurement of carcinoembryonic antigen levels in the cystic fluid is useful in checking for underdiagnosis of such tumors.

Primary retroperitoneal mucinous cystadenocarcinoma: report of a case.

A case of primary retroperitoneal mucinous cystadenocarcinoma is presented. Ultrasonography (US), computed tomography (CT), and magnetic resonance imaging (MRI) examinations demonstrated a large cystic tumor with solid excrescences. Angiographic examination demonstrated an avascular area with small areas of faint blush. It must be borne in mind that mucinous cystadenocarcinoma originating from the retroperitoneum can occur, although it is very rare.

Molecular typing of methicillin-resistant Staphylococcus aureus in a university teaching hospital.

Plasmid analysis and pulsed-field gel electrophoresis (PFGE) were used to study the epidemiologic relationship among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated at Tokyo Medical and Dental University Hospital. We found that 263 of 276 MRSA isolates had plasmids, which could be classified into 30 different patterns according to the number and plasmid molecular weight. Strains which harboured a single plasmid of approximately 13.4 Mds in molecular weight were the most numerous (55.7% of the isolates). These strains were isolated from 14 of 17 hospital wards. The largest number of strains with this plasmid pattern (33 strains) were isolated from a single ward. PFGE typing was then performed to further confirm the relationships among these 33 strains. The PFGE banding patterns of these strains were highly similar. The antibiogram profiles of these strains were also correlated with the PFGE pattern. Thus, the results suggest that these strains are epidemiologically related and spread throughout the ward. Combined plasmid analysis and PFGE were effective for discriminating the various MRSA isolates.

[In vitro antimicrobial activity of rokitamycin, a macrolide antibacterial agent, against clinically isolated strains of Campylobacter and other enteritis-causing bacteria].

We determined the minimum inhibitory concentrations (MICs) of rokitamycin (TMS-19-Q, RKM), a macrolide antimicrobial agent, against strains of various bacterial species isolated from enteritis patients, and compared them with those of josamycin (JM), erythromycin (EM) and ofloxacin (OFLX). MIC90 of RKM against 147 strains of Campylobacter jejuni, and each 25 strains of Shigella spp., Salmonella spp. and diarrheagenic Escherichia coli were 1.56, 200, 800 and 200 micrograms/ml, respectively. There was only one RKM resistant (MIC greater than 100 micrograms/ml) C. jejuni strain, while most of the strains of the other species were resistant to RKM. MIC values of the other drugs were all similar to those of RKM. MIC90 of OFLX against 147 strains of C. jejuni was 0.78 micrograms/ml, lower than other drugs.

[Characteristics of the Shigella flexneri strains isolated from Japanese monkeys (Macaca fuscata)].

Shigella flexneri was isolated from 11 of 95 Japanese monkeys (Macaca fuscata) purchased by the Animal Research Center, Tokyo Medical and Dental University, during the period from 1988 through 1990. The serotypes of the isolates were 4a (5 isolates), 2a (4 isolates), 2b (1 isolate), and varY (1 isolate). Although some of the strains in the same serotypes were similar, others were different in their antibiotic susceptibility patterns or the plasmid profiles. Our results suggested that various types of Shigella flexneri had infected the Japanese monkeys. However, the source(s) and the routes of infection were unclear. Dysentery in the Japanese monkey has not yet been reported. So, to our knowledge, this may be the first report of dysentery among Japanese monkeys.

[A study on stressful life events of workers in Japan].

The experience rates of eighteen life events of Japanese workers were surveyed and the strength of each of the events was evaluated by a self-reported questionnaire. The fifteen professions surveyed included construction, transport, mailing, chemical production, banking, newspaper, TV services, commercial publishing, advertising, teaching and civil service. The number of workers who answered was 18,657 males and 4,443 females, aged 20 to 59 years. The male workers were divided and analysed in six job groups: clerical workers (n = 5,866), professionals (n = 3,696), blue collar day workers (n = 1,623), blue collar night or shift workers (n = 3,191), drivers (n = 1,663) and construction workers (n = 2,466). They were divided into groups and compared according to job, sex and five different age groups. The highest experience rate in each of the life events such as family trouble for both sexes, death of a family member, financial trouble and anxiety, death of a close friend, dissatisfactory transport to workplace or job, respectively. Comparing the experience rates between males and females we found no significant difference for five items. However, other items had higher experience rates for males than for females with the exception of family trouble. In regard to the age characteristics of each of the life events, as the age increased the experience rates of health-related life events such as the death of a spouse, child, family member or close friend and one's own illness or injury became higher. In contrast, the experience rates of items such as moving to a worse residence and failure in a school or training program became lower as the age increased for both sexes. Among job groups, construction workers had the highest experience rates of most life events except for the item of dissatisfactory transport to the workplace or job. Among other job groups, drivers had higher experience rates in the following four items: re-employment, death of spouse, divorce and financial trouble and anxiety. Blue collar day workers had higher rates for failure in school or training program and family trouble. The blue collar night or shift workers had higher rates for failure in school or training program and dissatisfactory transport to the workplace or job. The mean values of strength in six life events: re-employment, death of a family member, death of a close friend, serious physical illness, dissatisfactory transport to the workplace or job and family trouble were stronger in females than in males.(ABSTRACT TRUNCATED AT 400 WORDS)

[Effect of DQ-2556, a new cephalosporin derivative, on fecal microflora].

A new cephalosporin derivative antibacterial agent, DQ-2556, was administered intravenously to 4 healthy adult male volunteers at 2 grams per dosage 2 times a day for 5 days, and degrees of effects of drug concentrations on the fecal microflora were investigated. 1. The total viable count remained unchanged during the study period due to the minimal change in the number of members of the family Bacteroidaceae, which were the most dominant organisms. In one of the volunteers, however, the total count was remarkably reduced immediately after the end of administration of the drug. 2. Among the obligate anaerobes, most of the organisms decreased except members of the family Bacteroidaceae. A marked increase of lecithinase-positive clostridia was noted in 2 volunteers. 3. The number of aerobes, including facultative anaerobes, members of the family Enterobacteriaceae were suppressed but bacilli and yeasts showed slight increases. In many cases, increases in enterococci were also observed. 4. DQ-2556 was not detected in the feces from any of the volunteers during the study period. 5. Neither Clostridium difficile nor its toxin D-1 was detected in any of the volunteers. 6. Diarrhoea or other side-effects were not noted in any of the volunteers.

The effect of levofloxacin, an optically-active isomer of ofloxacin, on fecal microflora in human volunteers.

Following oral administration of levofloxacin (LVFX, (S)-(-)-Ofloxacin; formerly designated as DR-3355) at 200 mg per dose 3 times a day for 7 days to 6 healthy male volunteers, degrees of disturbance of the fecal microflora and fecal drug concentrations were examined. The total viable count remained unchanged during the study period due to the minimal change in the number of members of the family Bacteroidaceae, the most predominant organisms. Most of the aerobes including facultative anaerobes were suppressed by LVFX with only a slight increase in yeasts. In particular, the members of the family Enterobacteriaceae were reduced to below the detection limit on and after day 3 through the time of discontinuation of the drug in all subjects but one. Among the obligate anaerobes, peptostreptococci and bifidobacteria decreased or disappeared in some volunteers, but no significant changes were observed in other anaerobes. Neither Clostridium difficile nor its toxin D-1 was detected in any of the volunteers. No side effects attributable to the drug were observed. During administration, LVFX was detected in the feces at high concentrations which correlated well with the decrease of susceptible members of flora as well as to their detection rate.

[The effect of new broad-spectrum cephamycin, cefotetan administered intravenously on human fecal flora (author's transl)].

The changes in the fecal flora and their correlation with the fecal drug concentrations were studied, employing 4 volunteers (healthy adult males) to whom 1 g of cefotetan was intravenously administered daily for 6 days. 1. In all of the subjects, the total counts of fecal organisms were slightly decreased 1 day after administration of cefotetan, then showed gradual increase, and recovered to the normal level on 8 days after the withdrawal of the drug. 2. Cefotetan caused drastic suppression of normal aerobic and anaerobic flora during the administration period, except streptococci that remained unchanged at all. The bacterial groups suppressed were bacteroidaceae, eubacteria, bifidobacteria, peptococcaceae, lactobacilli, enterobacteriaceae, lecithinase-positive clostridia, and staphylococci. Yeasts were found to increase during the administration period. 3. The changes in flora well correlated with the fecal level of the drug concentrations. This is in accordance with the in vitro antibacterial activity of the drug. 4. A period of more than 15 days was required after the withdrawal of cefotetan to recover the normal fecal flora. 5. Loss of body weight and diarrhoea were not observed in any of the subjects.

The effect of DU-6859a, a new potent fluoroquinolone, on fecal microflora in human volunteers.

Following the oral administration of DU-6859a to six healthy male volunteers at 100 mg per dose three times a day for seven consecutive days, the degree of disturbances of fecal microflora and fecal drug concentrations were examined. The total viable count decreased transiently during the administration due to changes in the number of members of the family Bacteroidaceae, the most predominant organisms. The other obligate anaerobes and most aerobes including facultative anaerobes were suppressed markedly by DU-6859a, with an exception of yeasts. Members of the family Enterobacteriaceae and most of anaerobic bacteria were reduced to below the detection limit in all subjects from the 3rd day of administration period to one day after the discontinuation of the drug. Clostridium difficile was detected in one of the volunteers at the end of administration. Side effects, such as mild transient soft stool and mild transient diarrhoea, were observed in five and one volunteers, respectively. During the administration, DU-6859a was detected in feces at high concentrations which correlated well with the decrease of susceptible members of fecal flora as well as their detection rate.

[Studies on the method of susceptibility test of Campylobacter jejuni to fosfomycin].

Conditions for the susceptibility test of Campylobacter jejuni to fosfomycin (FOM) were studied and it was concluded that the following procedure is most appropriate for routine tests. The test organisms were subcultured in 0.5 ml of heart infusion broth (Difco) in test tubes for a microplanter (Sakuma Seisakusho) and incubated in a GasPak jar with a CampyPak and catalyst (BBL) at 37 degrees C for 44--48 hours. The cultures were then diluted 100-fold in the same broth and inoculated with a microplanter on test plates. The test plates were nutrient agar (Difco) supplemented with 0.2% MgCl2 and 5% defibrinated horse blood. The plates were then incubated under the condition as described above and the growth of the cells was examined. It should be noted that drying of plates for more than 1 hour caused significant loss of viability of the cells.

[Electromyographic analysis on juvenile muscular atrophy of unilateral upper extremity (author's transl)].

In 1959, this disorder was first reported by one of authors (K.H.) (6). Since then, many Japanese cases have been reported and recently some cases are also reported in other countries. Its cause and pathology are still unknown, because no autopsy case was obtained. In this report, some electromyographic findings are described and discussed. This series contain 26 cases (male 23, female 3). Age of onset: 13-33 years old (mean 19.4 years old). Clinical duration: 2-32 years old (mean 9.0 years). Only 6 cases (23%) showed trauma as the past history (Table 2). We classified 26 cases in three groups (UL: unilateral, BP: bilateral, one-side predominant, BS: bilateral symmetrical) based on the laterality of physical findings (amyotrophy, weakness, finger tremor, etc.) (Table 3). Conventional needle EMG were done in all cases and these findings were judged by the criteria which we defined as Fig. 2. In some cases, conduction velocity (MCV & SCV) were measured. Needle EMG showed more selective and localized distribution of NGP (neurogenic pattern) in the hand muscles, the forearm muscles except for M. brachioradialis, and M. triceps brachii on the atrophied sides of the arm in all clinical groups (UL, BP, BS) (Table 4.5). On the normal side of bilaterally examined cases of UL group, NGP was shown in 9/11 cases, buts its distribution seemed to be slightly restricted in the more distal part. Thus, in this disorder, 22/26 cases (77%) were electromyographically affected on the bilateral sides, although the clinical feature was mostly unilateral. The clinical features and pathological process seemed to be established in a few years after the onset without the apparent tendency of further progression. Afterwards, no apparent extension of NGP distribution and increment of grading were observed in the Clinical course. No delay was observed on MCV and SCV. The pathological lesions responsible for these electromyographic changes are discussed, either on spinal segmental origin or on brachial plexus origin, but some ambiguous portions are still remained.

[The usefulness of nucleolar organizer regions in diagnosis of colonic epithelial neoplasia].

The Ag-NORs technique, staining nucleolar organizer regions, was performed on 23 areas of colonic normal mucosa, 17 adenomas with low grade atypical and 8 adenocarcinoma invading submucosa, to examine its possibility of application to the pathological diagnosis of colonic epithelial neoplasia. In normal colonic gland, the average number of Ag-NORs per 1 nucleus in cells of lower two-thirds showed significantly higher value (p less than 0.01) than higher one-third, and the mean cross-section area per 1 Ag-NOR in lower two-thirds showed a tendency to be larger than higher one-third. This results suggested that there was a positive correlation between the proliferative activity of cells and the number and size of Ag-NORs. A comparison of normal gland, adenoma and adenocarcinoma showed that the mean cross-section area of Ag-NORs correlated positively to their histological grade of atypia, but the average number did not. However using these two parameters, these three lesions had a tendency to be discriminated respectively at the average Ag-NORs number of 2.8 dots and the area of 2 microns. These results suggested that the Ag-NORs technique was expected to be a useful method for histopathological diagnosis of colonic epithelial neoplasia.