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.

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.

Molecular analysis of Clostridium difficile at a university teaching hospital in Japan: a shift in the predominant type over a five-year period.

Clostridium difficile isolates recovered from patients admitted to a teaching hospital in Japan over a 5-year period were analyzed. Two molecular typing systems, PCR ribotyping and pulsed-field gel electrophoresis (PFGE) analysis, were used. Twenty-six PCR ribotypes were found among the 148 isolates. The predominant type at our hospital appeared to shift during the study period, from PCR ribotype a in 2000 (15/33, 45%) to PCR ribotype f in 2004 (18/28, 64%). By using PFGE with thiourea added to both the gel and running buffer, all 148 Clostridium difficile isolates were successfully classified into 37 types and 61 subtypes. The PCR ribotype f isolates were further classified into four types and 11 subtypes by PFGE. The PFGE patterns of the 11 subtypes differed from each other by only 1 to 4 bands, suggesting that these differences might reflect genetic changes during patient-to-patient transmission over the 5-year period analyzed, and that PCR ribotype f isolates might be outbreak-related. In addition, the PCR ribotype f was identical to the PCR ribotype designated smz, which is reported to have caused multiple outbreaks in Japan. These results confirmed that PCR ribotype f (type smz) has specific virulence or survival factors that make it more likely to cause nosocomial outbreaks at Japanese hospitals. PCR ribotype 027, which has been reported to have caused recent outbreaks in North America and Europe, was recovered from one patient in this study; however, this strain was community-acquired. Our findings emphasize the importance of monitoring specific strains to control and prevent nosocomial infection.

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.

Pleuropericarditis and disseminated intravascular coagulation in ulcerative colitis.

We report a 30-year-old woman with pleuropericarditis, cardiac tamponade, and disseminated intravascular coagulation complicating active ulcerative colitis (UC). Other autoimmune diseases were not present. She responded to pulsed steroid therapy and anticoagulant with resolution of the complication and UC. We reviewed the literature and found 27 cases of pleuropericarditis associated with idiopathic inflammatory bowel disease (IBD). It has been reported that pleuropericarditis associated with IBD responds well to nonsteroidal antiinflammatory drugs, as well as steroids. The causes of cardiac involvement in IBD remain unclear, but the pleuropericarditis must be recognized as a potential extraintestinal manifestation of IBD.

The complete DNA sequence of the O antigen gene region of Plesiomonas shigelloides serotype O17 which is identical to Shigella sonnei form I antigen.

We cloned and determined the sequence of a DNA region of approximately 15-kb containing the cluster of genes required for O17 antigen expression in the Escherichia coli K-12 strain from the chromosome of Plesiomonas shigelloides serotype O17:H2 strain. The sequencing analysis revealed that the minimum essential region of the P. shigelloides O17 antigen gene cluster had a size of approximately 11.5-kb and contained 9 contiguous open reading frames (ORFs), which were almost identical to the corresponding ORFs of Shigella sonnei form I antigen gene region, except for IS630 sequence, at the DNA as well as amino acid levels. The putative function of most of the ORFs could be determined on the basis of amino acid sequence similarities and characteristics. In addition, the G+C content of the P. shigelloides O17 antigen genes was lower than that of the chromosomal DNA of P. shigelloides and S. sonnei, suggesting that both P. shigelloides O17 and S. sonnei form I antigen genes had been derived from the same origin with a low G+C content.

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.

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.

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.

Light and electron microscopy of stage-specific features of the transdifferentiation of mouse Meckel's cartilage chondrocytes in vitro.

The changes in the morphological characteristics of Meckel's cartilage cells in culture can be divided into five stages during their transdifferentiation to cells with an osteocyte-like phenotype, as demonstrated in this study by light and electron microscopy. Chondrocytes were isolated from day-17 embryonic mice by enzymatic digestion and inoculated at a density of 1 x 10(4) cells. The cultures were incubated at 37 degrees C and cells were harvested every week for 4 weeks and processed for light and electron microscopy, as well as an immunohistochemical analysis of type I and II collagens. The primary cultures were characterized by fibroblastic cells (stage 1) around days 2-4. Polygonal cells (stage 2) expanded on a poorly formed extracellular matrix on day 7, and then they differentiated into large, round cells (stage 3) which began to form cellular nodules. The large, round cells contained many vacuoles, which were rapidly released into the pericellular space with a significant resultant decrease in cell size. Immunostaining showed that the extracellular matrix was occupied predominantly by type II collagen after 2 weeks in culture. However, the type II collagen was gradually replaced by type 1 collagen. Cells that formed multiple layers gradually changed into small, round cells (stage 4) at the tops of nodules and ultimately expressed an osteocytic phenotype (stage 5) after further transdifferentiation.

Early postoperative evaluation of pylorus-preserving gastrectomy for gastric cancer.

Early postoperative evaluation was prospectively performed in 35 gastric cancer patients after pylorus-preserving gastrectomy (PPG) between 1989 and 1991, comparing the results with those of 29 patients who underwent conventional distal gastrectomy (CDG). Surgical stress, including the duration of operation (149.0 +/- 4.3 minutes) and the total volume of bleeding at operation (97.0 +/- 11.2 g), was significantly less in the PPG patients. Early postoperative complications were seen in 31% after PPG and in 35% after CDG. The most frequent complication in PPG patients was remnant gastric stasis (23%). Endoscopy showed redness or erosion (or both) of the gastric remnant in 17% after PPG and in 81% after CDG. Bile regurgitation was demonstrated in 11% after PPG and in 62% after CDG. In PPG patients, the pyloric ring opened and closed during the examination. Gastric pH was 4.2 +/- 0.4 in PPG patients but was significantly lower in CDG patients. The resting gallbladder area, examined by ultrasonography, demonstrated no changes after PPG but was significantly enlarged after CDG (from 11.3 +/- 1.2 cm2 to 15.8 +/- 1.5 cm2 at 2 weeks). The percentage of the original resting gallbladder area at 20 minutes after injection of cerulein increased slightly in PPG patients but recovered thereafter, whereas in CDG patients it increased significantly (from 39.4 +/- 8.3% to 66.7 +/- 9.1% at 2 weeks). No gallstone formation was detected throughout the observation period after PPG, whereas after CDG it was detected in two patients at 1 year.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

[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)

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.

[Mental health and its related factors among middle-aged male workers: an eighteen-month follow-up study].

Based on a longitudinal questionnaire survey of 11,121 subjects (males aged 30-59, engaged in various occupations and firms), this study examined the relationships of the baseline psychosocial work environment and personal health practices to mental disorders which occurred during an 18-month follow-up period. The onset of mental disorders during the follow-up period was assumed to be the time when the subject reported to have received medical treatment for the first time due to mental disorder. The psychosocial factors of the work environment were conceptually constructed from two aspects of working conditions and occupational stress. Working conditions were assessed by actual working hours, frequencies of late night work (10 p.m.-5 a.m.), and role in the organization. Occupational stress was measured by psychological work overload and job characteristics (job demands, decision latitude). Various factors associated with positive health behavior were examined through an index based on six variables, that is, supper time, exercise, smoking, alcohol consumption, sleep and weight. Multiple logistic analysis showed that both psychological work overload and late night work have a significant independent effect on the onset of mental disorders after controlling for other variables.

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.

[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.

Chemical analysis of lipopolysaccharides of Shigella sonnei form II strains expressed by cloned form I antigen genes.

A compositional sugar analysis was carried out on lipopolysaccharide (LPS) from Shigella sonnei form II in which a plasmid with cloned form I antigen genes had been introduced. The recipient form II strains contained galactose, glucose, heptose, glucosamine, and 2-keto-3-deoxyoctonic acid (KDO) (2: 3: 1: 2: 2) in its LPS, while the transformant form I LPS contained, besides these sugars, N-acetyl-L-altrosaminouronic acid as an additional sugar constituent, which is known to be one of the antigenic determinants of form I antigen.