High production of a class III lantipeptide AmfS in Streptomyces griseus.

AmfS, a class III lantipeptide serves as a morphogen in Streptomyces griseus. Here, we constructed a high production system of AmfS in S. griseus. We isolated S. griseus Grd1 strain defective in glucose repression of aerial mycelium formation and found it suitable for the overproduction of AmfS. Two expression vectors carrying the strong and constitutive ermE2 promoter were constructed using a multicopy number plasmid, pIJ702. The use of the Grd1 strain combined with the expression vectors enabled high production of AmfS by S. griseus into its culture broth. The expression system was also effective for the generation of abundant AmfS derived from Streptomyces avermitilis. In addition, site-directed mutagenesis revealed the amino acid residues essential for the morphogen activity of AmfS. These results indicate that the constructed system enables efficient production of class III lantipeptides by Streptomyces.

[Bacteria isolated from surgical infections and its susceptibilities to antimicrobial agents--special references to bacteria isolated between April 2009 and March 2010].

Bacteria isolated from surgical infections during the period from April 2009 to March 2010 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 671 strains including 16 strains of Candida spp. were isolated from 174 (79.1%) of 220 patients with surgical infections. Four hundred and eleven strains were isolated from primary infections, and 244 strains were isolated from surgical site infection. From primary infections, anaerobic Gram-negative bacteria were predominant, followed by aerobic Gram-negative bacteria, while from surgical site infection aerobic Gram-positive bacteria were predominant, followed by anaerobic Gram-negative bacteria. Among aerobic Gram-positive bacteria, the isolation rate of Enterococcus spp. was highest, followed by Streptococcus spp., and Staphylococcus spp. in this order, from primary infections, while Enterococcus spp. was highest, followed by Staphylococcus spp. from surgical site infection. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae, Enterobacter cloacae and Pseudomonas aeruginosa, in this order, and from surgical site infection, E. coli was most predominantly isolated, followed by P. aeruginosa and E. cloacae. Among anaerobic Gram-positive bacteria, the isolation rate of Eggerthella lenta was the highest from primary infections, followed by Parvimonas micra, Streptococcus constellatus and Finegoldia magna, and from surgical site infection, E. lenta was most predominantly isolated. Among anaerobic Gram-negative bacteria, the isolation rate of Bilophila wadsworthia was the highest from primary infections, followed by Bacteroides fragilis, Bacteroides ovatus and Bacteroides thetaiotaomicron, and from surgical site infection, B. fragilis was most predominantly isolated, followed by B. ovatus, B. wadsworthia and B. thetaiotaomicron, in this order. In this series, we noticed no vancomycin-resistant Gram-positive cocci, nor multidrug-resistant P. aeruginosa. We should carefully follow up B. wadsworthia which was resistant to various antibiotics, and also Bacteroides spp. which was resistant to many beta-lactam antibiotics.

[Bacteria isolated from surgical infections and their susceptibilities to antimicrobial agents --special references to bacteria isolated between April 2008 and March 2009].

Bacteria isolated from infections in abdominal surgery during the period from April 2008 to March 2009 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 712 strains including 18 strains of Candida spp. were isolated from 173 (80.5%) of 215 patients with surgical infections. Three hundred and sixty-six strains were isolated from primary infections, and 346 strains were isolated from postoperative infections. From primary infections, anaerobic Gram-negative bacteria were predominant, followed by aerobic Gram-negative bacteria, while from postoperative infections aerobic Gram-positive bacteria were predominant, followed by anaerobic Gram-negative bacteria. Among aerobic Gram-positive bacteria, the isolation rate of Enterococcus spp. was highest, followed by Streptococcus spp., and Staphylococcus spp. in this order, from primary infections, while Enterococcus spp. was highest, followed by Staphylococcus spp. from postoperative infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae and Pseudomonas aeruginosa, in this order, and from postoperative infections, P aeruginosa was most predominantly isolated, followed by E. coli, Enterobacter cloacae, and K. pneumoniae. Among anaerobic Gram-positive bacteria, the isolation rate of Eggerthella lenta was the highest from primary infections, followed by Parvimonas micra, Streptococcus constellatus and Gemella morbillorum, and from postoperative infections, E. lenta was most predominantly isolated. Among anaerobic Gram-negative bacteria, the isolation rate of Bacteroides fragilis was the highest from primary infections, followed by Bacteroides thetaiotaomicron, Bacteroides ovatus and Bilophila wadsworthia, and from postoperative infections, B. fragilis was most predominantly isolated, followed by B. thetaiotaomicron, B. wadsworthia and B. ovatus, in this order. In this series, we noticed no vancomycin-resistant methicillin-resistant S. aureus, and Enterococcus spp., nor multidrug-resistant P aeruginosa. We should carefully follow up B. wadsworthia which was resistant to various antibiotics, and also Bacteroides spp. which was resistant to many beta-lactam antibiotics.

[Bacteria isolated from surgical infections and their susceptibility to antimicrobial agents--special reference to bacteria isolated between April 2007 and March 2008].

Bacteria isolated from infections in abdominal surgery during the period from April 2007 to March 2008 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 707 strains including 24 strains of Candida spp. were isolated from 181 (79.0%) of 229 patients with surgical infections. Three hundred and ninety-five strains were isolated from primary infections, and 288 strains were isolated from postoperative infections. From primary infections, anaerobic Gram-negative bacteria were predominant, followed by aerobic Gram-negative bacteria, while from postoperative infections aerobic Gram-positive bacteria were predominant, followed by anaerobic Gram-negative bacteria. Among aerobic Gram-positive bacteria, the isolation rate of Enterococcus spp. was highest, followed by Streptococcus spp., and Staphylococcus spp. in this order, from primary infections, while Enterococcus spp. was highest, followed by Staphylococcus spp. from postoperative infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterobacter cloacae, in this order, and from postoperative infections, P. aeruginosa was most predominantly isolated, followed by E. cloacae, E. coli and K. pneumoniae. Among anaerobic Gram-positive bacteria, the isolation rate of Parvimonas micra was the highest from primary infections, followed by Streptococcus constellatus and Gemella morbillorum, and from postoperative infections, Anaerococcus prevotii was most predominantly isolated. Among anaerobic Gram-negative bacteria, the isolation rate of both Bacteroides fragilis and Bilophila wadsworthia were the highest from primary infections, followed by Bacteroides thetaiotaomicron and Campylobacter gracilis, and from postoperative infections, B. thetaiotaomicron was most predominately isolated, followed by B. fragilis, Bacteroides caccae and B. wadsworthia in this order. In this series, we noticed no vancomycin-resistant Gram-positive cocci, nor multidrug-resistant P aeruginosa. There were nine strains of coagulase-negative Staphylococci which show higher MIC against teicoplanin more than 4 gg/mL, but all of them had good susceptibilities against various anti-MRSA antibiotics. We should carefully follow up B. wadsworthia which was resistant to various antibiotics, and also Bacteroides spp. which was resistant to many beta-lactam antibiotics.

[Bacteria isolated from surgical infections and its susceptibilities to antimicrobial agents--special references to bacteria isolated between April 2006 and March 2007].

Tendency of isolated bacteria from infections in abdominal surgery during the period from April 2006 to March 2007 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 474 strains including 23 strains of Candida spp. were isolated from 170 (75.2%) of 226 patients with surgical infections. Two hundred and twenty-six strains were isolated from primary infections, and 224 strains were isolated from postoperative infections. From primary infections, anaerobic Gram-negative bacteria were predominant, followed by aerobic Gram-negative bacteria, while from postoperative infections aerobic Gram-positive bacteria were predominant, followed by anaerobic Gram-negative bacteria. Among aerobic Gram-positive bacteria, the isolation rate of Staphylococcus spp. was higher from postoperative infections, while Enterococcus spp. was higher from primary infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae, Enterobacter cloacae and Pseudomonas aeruginosa, in this order, and from postoperative infections, P. aeruginosa was the most predominantly isolated, followed by E. coli and E. cloacae. Among anaerobic Gram-negative bacteria, the isolation rate of Bilophila wadsworthia was the highest from primary infections, followed by Bacteroides fragilis and from postoperative infections, B. fragilis was most predominately isolated, followed by Bacteroides caccae, Bacteroides thetaiotaomicron and B. wadsworthia in this order. In this series, we noticed no methicillin-resistant Staphylococcus aureus, nor multidrug-resistant P. aeruginosa. There were three strains of methicillin-resistant coagulase-negative Staphylococcus aureus, but all of them had good susceptibilities against various anti-MRSA antibiotics. We should carefully follow up B. wadsworthia.

[Bacteria isolated from surgical infections and its susceptibilities to antimicrobial agents--special references to bacteria isolated between april 2003 and march 2004].

Tendency of isolated bacteria from infections in abdominal surgery during the period from April 2005 to March 2006 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 384 strains including 18 strains of Candida spp. were isolated from 161 (70.3%) of 229 patients with surgical infections. One hundred and ninty-five strains were isolated from primary infections, and 171 strains were isolated from postoperative infections. From primary infections, aerobic Gram-negative bacteria and aerobic Gram-positive bacteria were predominant, while aerobic Gram-positive bacteria were predominant from postoperative infections. The isolation rate of aerobic Gram-positive bacteria, such as Enterococcus spp. and Staphylococcus aureus were higher from both types of infections. Among anaerobic Gram-positive bacteria, the isolation rate of Peptostreptococcus spp. was the highest from both types of infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Pseudomonas aeruginosa, Klebsiella spp. in this order, and from postoperative infections, E. coli was the most predominantly isolated, followed by Klebsiella pneumoniae and P. aeruginosa. Among anaerobic Gram-negative bacteria, the isolation rate of Bacteroides fragilis group was the highest from both primary and postoperative infections. In this series, we noticed no vancomycin-resistant Gram-positive cocci, nor multidrug-resistant P. aeruginosa. But cefazolin-resistant E. coli producing extended spectrum fl-lactamase was seen in 5.0 per cents. We should be carefully followed up the facts that the increasing isolation rates of B. fragilis group and Bilophila wadsworthia which were resistant to both penicillins and cephems.

Treatment of peritoneal carcinomatosis using carboplatin-loaded hydroxyapatite particles.

UNLABELLED: The anti-tumor effects and tissue distribution of carboplatin (CBDCA) incorporated into hydroxyapatite (HAP) particles was studied. MATERIALS AND METHODS: Seven days after the intraperitoneal (i.p.) implantation of AH130 tumor cells into Donryu rats, the animals were randomized into four groups: group I was treated with saline i.p.; Group II, with CBDCA i.v.; group III with CBDCA i.p.; and group IV with HAP-CBDCA i.p. RESULTS: The survival rate of group IV was better than that of the other groups (p < 0.05). The area under the ascitic platinum concentration-time curve and tissue concentrations of platinum in the omentum after 24 hours of treatment were higher in group IV than in groups II or III (p < 0.05). The platinum concentrations in the kidneys of group IV were lower than in group III (p < 0.05). CONCLUSION: The HAP-CBDCA combination enhances the anti-tumor effects of the drug and reduces the nephrotoxicity in rats with peritoneal carcinomatosis.

[Prophylactic effect of fosfomycin on postoperative infection in gastroenterological surgery].

Fosfomycin (FOM), a drug with unique mechanism of action against bacteria, was prospectively compared with cefmetazole (CMZ) and flomoxef (FMOX) to assess its effectiveness in preventing postoperative infections after upper gastroenterological and hepatobiliary surgery. FOM (2g/dose), CMZ (1 g/dose), or FMOX (lg/dose) was infused intravenously 2-3 times daily for basically 4 days including the day of surgery in order to examine whether or not infections would develop after surgery. According to the analysis results of 162 cases, efficacy in preventing infection was 86.8% for FOM, 73.1% for CMZ and 83.3% for FMOX, showing no significant differences among the 3 groups. Thus FOM was considered as efficacious as CMZ or FMOX in prophylaxis against postoperative infections. There were no adverse reactions in the FOM or CMZ group, but 3 cases of adverse reactions occurred in the FMOX group. Based on the above results, the usefulness of FOM for prophylaxis against postoperative infections has been confirmed. At present, the first- and second-generation cephems are most frequently used for prophylaxis against postoperative infections. In order to prevent an increase of resistant strains due to inappropriate use of these drugs, FOM, having no cross resistance with currently used drugs, is a promising candidate for prophylaxis against postoperative infections.

[Successfully resected advanced gastric carcinoma after combined chemotherapy of TS-1 plus CDDP--a case report].

A 66-year-old man complained of epigastralgia and a close examination was done. A type 3 gastric carcinoma was found. The tumor invading the pancreas and swelling lymphnodes around the stomach were confirmed according to abdominal CT. We considered it difficult to resect the tumor completely, and so we used combined chemotherapy of TS-1 plus CDDP. TS-1 was taken at 120 mg/day for 28 days followed by 14 days rest. CDDP was injected at 140 mg on day 8. Because the tumor in the stomach changed to scar and swelling lymphnodes diminished after two courses of therapy, total gastrectomy was done. After operation, combined chemotherapy of LV and 5-FU resulted in no recurrence to date. Although combined chemotherapy of TS-1 plus CDDP is effective for advanced gastric carcinoma, the number of days TS-1 is taken and the duration of the preoperative chemotherapy must be investigated in forthcoming study.

Retrospective study on thymidylate synthase as a predictor of outcome and sensitivity to adjuvant chemotherapy in patients with curatively resected colorectal cancer.

We carried out a retrospective evaluation of thymidylate synthase (TS) expression in tumor tissue, and its relation to outcome and response to treatment. The treatment consisted of chemotherapy with tegafur and uracil (UFT). The study group comprised 245 patients with curatively resected Dukes' stage B or C colorectal cancer who were postoperatively enrolled in a controlled study and assigned to receive UFT or no adjuvant chemotherapy. TS expression in tumor tissue was evaluated immunohistochemically with the use of recombinant human TS-specific antibody. Results were as follows. There was no relation between TS expression and the rate of 5-year disease-free survival. Similar results were obtained in both colonic and rectal tumors. The rate of 5-year disease-free survival was significantly higher in the UFT group than in the group receiving no adjuvant chemotherapy ( =0.0055). The difference in survival became more marked among patients whose tumors had diffuse TS expression ( =0.0027). There was no difference in survival between the treatment groups among patients whose tumors had focal TS expression. We conclude that, although unrelated to outcome, TS activity may be useful in predicting the response to adjuvant chemotherapy with UFT in patients with curatively resected Dukes' stage B or C colorectal cancer.