Single nucleotide polymorphism of TAG-1 influences IVIg responsiveness of Japanese patients with CIDP.

OBJECTIVE: Chronic inflammatory demyelinating polyneuropathy (CIDP) is characterized by immune-mediated peripheral demyelination. Although corticosteroid, IV immunoglobulin (IVIg) and plasma exchange have been established as the most effective therapeutics, subpopulations of patients show little or no response to either of these therapies. In this study, we examined whether particular genetic factors influence the therapeutic responsiveness of patients with CIDP. METHODS: One hundred Japanese patients categorized as responders or nonresponders to IVIg therapy participated in our study. We performed an association analysis with single nucleotide polymorphisms (SNPs) and haplotype studies between the IVIg responders and nonresponders. RESULTS: Two separate SNPs, corresponding to TAG-1 (transient axonal glycoprotein 1) and CLEC10A (C-type lectin domain family 10, member A), showed strong significant differences between responders and nonresponders. Haplotype analysis of a series of expanded SNPs, from TAG-1 or CLEC10A, showed that only TAG-1 included a significant haplotype within 1 linkage disequilibrium block, which accommodates IVIg responsiveness. Diplotype analysis of TAG-1 also supported this observation. CONCLUSIONS: Transient axonal glycoprotein 1 is a crucial molecule involved in IV immunoglobulin responsiveness in Japanese patients with chronic inflammatory demyelinating polyneuropathy.

Lysine decarboxylase of Vibrio parahaemolyticus: kinetics of transcription and role in acid resistance.

AIM: The aim of this study was to investigate the detailed mechanisms of acid resistance in Vibrio parahaemolyticus. METHODS AND RESULTS: All 11 strains of V. parahaemolyticus survived lethal acidic conditions following acid adaptation, and accumulation of cadaverine was detected. The addition of lysine improved survival, suggesting that lysine decarboxylase plays a role in the adaptive acid tolerance response. Two open reading frames (ORF) in V. parahaemolyticus, which are separated by a noncoding region, were found to be highly homologous to bacterial lysine decarboxylase (cadA) and lysine/cadaverine antiporter (cadB) genes. Transcriptional analyses of this operon revealed acid induction and enhanced induction by external lysine. The relative expression ratio of each transcript was found to follow the trend of cadA mRNA > cadB mRNA > cadBA bi-cistronic mRNA. A mutated strain, with a disrupted cadA gene, showed attenuated acid survival. CONCLUSIONS: We identified the lysine decarboxylase gene operon of V. parahaemolyticus. Expression of this operon was induced under acidic conditions. The cadA-mutated strain constructed in this study showed weaker tolerance to acidic conditions than the wild-type strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Vibrio parahaemolyticus utilizes the lysine decarboxylation pathway for survival in acidic conditions.

[Increasing fluoroquinolone low-sensitivity in enterotoxigenic Escherichia coli isolated from diarrhea of overseas travelers in Tokyo].

Drug resistance trends were investigated for 1,318 enterotoxigenic Escherichia coli (ETEC) isolated from overseas traveler's diarrheal cases in Tokyo during 1988-1999. A total of 1.6% (21 strains) were nalidixic-acid resistant and fluoroquinolones (NFLX, OFLX, CPFX, LVFX, TFLX, SPFX; FQ) low-sensitive (or low-level-resistant). None of the strains were high-level-resistant to FQ. The FQ low-sensitive strains were isolated in 1996 for the first time, and increased from 3.4% in 1996 to 15.8% in 1999. Countries visited by travelers with the FQ low-sensitive ETEC were India (16 cases), Nepal (3 cases), Cambodia (1 case), and Egypt (1 case). Drug resistance-patterns of the FQ low-sensitive strains, including other drugs (CP, TC, SM, KM, ABPC, ST, NA, and FOM) tested, varied among the 6 types. Among those, multidrug resistant strains accounted for 57.1% (12 strains). The enterotoxin producing types of strains were LT (4 strains), ST (10 strains), and both (7 strains). The serotypes of the strains were classified into 16 types. The quinolone resistance determining regions (QRDRs) of the gyrA genes of the FQ low-sensitive strains were sequenced. The mutations of a Ser to a Leu at position 83 (Ser-83-->Leu) was found in 19 strains, and Asp-87-->Tyr was found in 2 strains.

[The trends of Vibrio parahaemolyticus foodborne outbreaks in Tokyo: 1989-2000].

The total number of foodborne outbreaks due to Vibrio parahaemolyticus in Tokyo during the last 12 years between 1989 and 2000 were 710. The number of outbreaks in a year was 55 in 1989, 75 in 1990, and there was a gradual decrease to 24 outbreaks in 1993 which was the smallest number during those 12 years. After 1994, the number of outbreaks increased dramatically year by year until 1998 (107 outbreaks). Then they had decreased slightly to 74 in 1999, 65 in 2000. The monthly incidence of V. parahaemolyticus foodborne outbreaks showed a peak in August (44.2%) each year. In the last 12 years, 88.7% of V. parahaemolyticus foodborne outbreaks occurred during the 3 months between July and September, while 99.9% occurred between June and October. The most prevalent serotype of V. parahaemolyticus also changed, the most prevalent was O4:K4 in 1989, O4:K8 in both 1990 and 1991, O1:K56 in 1992, and O4:K8 from 1993 through 1995. Serotype O3:K6 became the most prevalent in 1996 and has remained so to date. In addition, the new serotype O4:K68 had also appeared in 1998. The number of outbreaks due to serotype O4:K68 followed that of O3:K6. Thus, the trends of V. parahaemolyticus foodborne outbreaks during the last 12 years in Tokyo showed various characteristics and dramatic changes in causal organisms.

A study of quality assessment in clinical microbiology performance of independent laboratories in Tokyo: 18-year participation in the Tokyo Metropolitan Government External Quality Assessment Program.

We evaluated the performance of independent microbiology laboratories in Tokyo over an 18-year period of participation in the external quality assessment (EQA) program, and we estimated the impact of the EQA program. The study design was a longitudinal retrospective analysis of performance, including isolation, identification, and antimicrobial susceptibility testing of bacteria from simulated patients' samples, in "open" surveys compared with "blind" surveys. Independent microbiology laboratories, licensed by the Tokyo Metropolitan Government, have been subject to mandatory evaluation by the EQA program since 1982. Survey reports, correspondence, annual guidance meetings, and inspections are used as quality improvement strategies. The performance for identification in "blind" surveys was significantly worse than that in "open" surveys (P < 0.001). Poorly performing laboratories had common features, including inadequate supervision by physicians and lack of familiarity with the impact of variations on the use of the standards. However, there were improvements in the performance of identification of some pathogens. The performance of susceptibility testing has not yet reached the relatively high level seen for identification. Some of the smaller laboratories have been gradually acquired by commercial chains operating outside Tokyo. The EQA program has established a role both in regard to laboratory improvement and as an educational tool. However, the program lags behind these of other developed countries in regard to the practical sciences. The main problems in regard to laboratory improvements are a shortage of human resources in clinical microbiology, lack of standardization of laboratory methods, and the pressures of financial constraints in the Japanese medical insurance system.

Flavor production from a non-stick oil by moulds.

Natural flavor was accidentally produced from rice cake products in Japan. A non-stick oil had been sprayed on the products during the production process. It was found that a Penicillium corylophilum strain, a contaminant of the oil, produced the flavor from the oil. The ingredients of the flavor were four volatile substances, 2-heptanone, 2-nonanone, 2-heptanol, and 2-nonanol. Challenge tests with the mould strain in a rice cake system were performed under various conditions. The volatile substances were produced in the largest amounts at 25 degrees C, followed by 20 or 30 degrees C then 10 degrees C. 2-Heptanone was produced most remarkably at 25 degrees C, followed by 2-nonanone, 2-heptanol, and 2-nonanol. The growth patterns of the mould were similar between 20-30 degrees C, and the growth at 10 degrees C was delayed. The non-stick oil itself had neither flavor nor volatile substance. The flavor was also produced from coconut oil, which was one of the materials of the non-stick oil. No bacteria or yeasts tested produced any flavor from the non-stick oil, whereas most of the moulds tested produced flavor components.

[Serovar-distribution and drug-resistance of Salmonella strains isolated from domestic and imported cases during 1995-1999 in Tokyo].

A total of 2,277 non-typhoidal Salmonella strains consisting of 1,807 domestic strains and 470 imported strains isolated from sporadic cases during 1995-1999 in Tokyo, were examined regarding their serovar-distibution and their drug-resistance. The serological typing results showed that the domestic strains were classified into 17 O-groups and 99 serovars, and the imported strains were classified into 12 O-groups and 58 serovars. Among the serovars identified, Salmonella serovar Enteritidis (S. Enteritidis), S. Thompson, S. Hadar, S. Infantis, S. Typhimurium, and S. Litchfield were predominant in the domestic strains, whereas S. Enteritidis, S. Anatum, S. Hadar, and S. Weltevreden were predominant in the imported strains. The drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOM, and NFLX) showed that 34.0% of the domestic strains and 33.0% of the imported strains were resistant to any of the drugs examined. The serovars of a high resistant rate during this period were S. Blockley (100%), S. Hadar (96.6%), S. Typhimurium (63.6%), and S. Enteritidis (62.2%) in the domestic strains and S. Blockley (100%), S. Hadar (97.1%), S. Rissen (88.9%), S. Emek (83.3%), S. Panama (83.3%), and S. Typhimurium (77.8%) in the imported strains. Drug-resistance patterns of the resistant isolates varied to 60 types. Prevalent patterns recognized were SM, TC.SM, TC, TC.SM.KM.ST, TC.SM.KM, and CP.TC.SM.ABPC in the domestic strains and TC.SM, TC, NA, TC.SM.KM.NA, and TC.SM.NA in the imported strains.

[Analysis for microbial contamination in production of Japanese-style confectionery "Monaka"].

Food hygiene in Japanese-style confectionery factories is hard to practice because the businesses are small. In a supporting system of voluntary-based hygienic management in this field, we microbiologically investigated the production processes of "Monaka" in a workshop in Tokyo. We microbiologically assessed the processing environments as well as the products in the workshop, then proposed some improvements in the production of the confectionery. After the improvements, microbial contamination of the processing environments was reduced and no microbial contamination was found in the sugared bean, or "An" produced, though the product "Monaka" was still contaminated, especially by molds. It was clarified that the molds came from contaminated baked wheat shells, or "Kawa" and further that the wheat shells were contaminated by molds during storage.

[Species and serovar-distribution, and drug-resistance of Shigella strains isolated from imported and domestic cases during 1995-1999 in Tokyo].

A total of 290 Shigella strains consisting of 180 imported strains and 110 domestic strains isolated during 1995-1999 in Tokyo were examined regarding their species and serovar-distribution and their drug-resistance. In both groups, S. sonnei (70.0% in the imported strains, 80.9% in the domestic strains) was found to be the most prevalent species, followed by S. flexneri (20.0% in the imported strains, 19.1% in the domestic strains). S. dysenteriae and S. boydii were only isolated in the imported cases. Among the S. flexneri serovar, 1b, 2a, 6, 2b, and 3a were predominant in the imported strains, whereas 1b and 2a were predominant in the domestic strains. Provisional new serovar Shigella strains were isolated from 11 imported cases and 2 domestic cases. The drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOM, and NFLX) showed that 92.2% of the imported strains and 94.5% of the domestic strains were resistant to some of the drugs tested. Drug-resistance patterns of the resistant strains varied in 25 types. Among those, a triple drug-resistance type with TC.SM.ST was found as the most frequent pattern in both groups. None of the strains were resistant to NFLX.

Comparison of capillary and test tube procedures for analysis of thermal inactivation kinetics of mold spores.

Characteristics of capillary and test tube procedures for thermal inactivation kinetic analysis of microbial cells were studied for mold spores. During heating, capillaries were submerged in a water bath and test tubes were held with their caps positioned above the level of the heating medium. Thermal inactivation curves of Aspergillus niger spores in capillaries at around 60 degrees C consisted of a shoulder and a fast linear decline, whereas curves in test tubes consisted of a shoulder, a fast linear decline, and a horizontal tail. There were no significant differences in values of the rate and the delay of fast declines in curves between the procedures. Some experiments were done to clarify the cause for tailing with test tubes. There were no tails with test tubes whose inner walls were not contaminated by A. niger spores, suggesting that tails arise from A. niger spores contaminating the inner walls of test tubes. Temperature of the inner wall at the level of a heating medium was lower than that of the medium. Further, there were no tails for test tubes submerged in the heating medium. These results showed that the reason for survival of contaminants on the upper wall of test tubes was that cells were not subjected to sufficient inactivation temperature. Finally, thermal inactivation curves of A. niger spores in capillaries at various constant temperatures were studied. Curves consisted of a shoulder and a fast linear decline at 57 degrees C and above, whereas curves at below 57 degrees C consisted of a shoulder, a fast linear decline, and a sloping tail.

[Increasing nalidixic-acid-resistance in Salmonella strains isolated from sporadic cases in Tokyo].

A total of 118 nalidixic-acid (NA)-resistant Salmonella strains consisting of 68 domestic strains and 50 imported strains isolated during 1988-1998 in Tokyo were examined regarding their annual incidence, serovars, drug-resistance patterns, and minimum inhibitory concentrations(MIC) to fluoroquinolones (NFLX, OFLX, ENX, and CPFX). NA-resistant strains accounted for 1.3% of all Salmonella (5,302 strains) isolated from domestic cases, and 2.5% of all Salmonella (1,981 strains) isolated from imported cases. The incidence of NA-resistant strains has increased since 1995, and it has been remarkable in imported cases. The results of the serotyping showed that the NA-resistant strains were classified into 25 serovars, excluding untypable strains. Among those, S. Enteritidis (21 strains), S. Blockley (13 strains), S. Litchifield (13 strains), S. Typhimurium (13 strains), S. Hadar (9 strains), and S. Virchow (8 strains) were predominant. Drug-resistance patterns of NA-resistant strains, including other drugs (CP, TC, SM, KM, ABPC, ST, FOM, and NFLX) tested varied among the 26 types. Among those, multidrug-resistant strains accounted for 61.9% (73 strains), and one strain among them was high-resistant to NFLX. MIC distribution of NA-resistant strains to fluoroquinolones showed that the ranges of all drugs were 4-128 times higher than NA-sensitive strains used for controls.

[Strains of Shigella sonnei recently isolated in Tokyo].

A total of 341 Shigella sonnei strains consisting of 94 domestic strains and 247 imported strains isolated during 1990-1997 in Tokyo, were examined regarding their colicine-type, drug-resistance and ornithine-utilization. The colicine typing results showed that the domestic strains were classified into 7 types, and the imported strains were classified into 13 types. Among the colicine-types identified, 8-type, 0-type, 6-type and 12-type were predominant in the domestic strains, whereas 6-type, 0-type, 8-type, 9A-type and 12-type were predominant in the imported strains. The drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOM and NFLX) showed that 89.4% of the domestic strains and 85.4% of the imported strains were resistant to some of the drugs except FOM and NFLX. Drugs with a high resistant rate were SM, TC and ST for both groups. Drug-resistance patterns of the resistant strains varied in 22 types. Among those, a triple drug-resistance type with TC.SM.ST was found in the most frequent pattern in both groups. The results of the ornithine-utilization test revealed that 28.7% of the domestic strains and 8.1% of the imported strains were negative. The ornithine-negative strains in the same source had a similar plasmid-profile, but generally there was no correlation between the different sources.

Growth of moulds inoculated into commercial mineral water.

The growth of mould spores of Penicillium sp. and Cladosporium sp. inoculated in a commercial mineral water product was studied. The strains had been isolated as fungal foreign bodies in commercial mineral waters. In product A, which was not originally sterilized and was contaminated with psychrophilic bacteria, the inoculated mould spores of the strains did not grow; no increases in viable colony counts or beta-glucans concentration in the samples were observed during storage. In a sterilized product A, inoculated spores of the strains grew into visible foreign bodies. The viable colony counts and the beta-glucans concentration in the samples increased during storage. These results showed that in a sterilized mineral water product, mould spores could grow into visible foreign bodies.

[Drug-resistance and definitive type 104 of Salmonella serovar typhimurium isolated from sporadic cases in Tokyo, 1980-1998].

A total of 674 Salmonella serovar Typhimurium (S. Typhimurium) strains consisting of 522 domestic strains and 152 imported strains isolated in Tokyo, 1980-1998, were examined regarding their drug-resistance and phage-type. Domestic strains accounted for 6.2% of all Salmonella (8,359 strains) isolated from domestic cases, and imported strains accounted for 3.7% of all Salmonella (4,083 strains) isolated from imported cases. A drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOM, and NFLX) showed that 245 strains (46.9%) of the domestic strains and 109 strains (71.7%) of the imported strains were resistant to some of the drugs, excluding FOM and NFLX. Drugs with a high resistance rate were TC, SM, ABPC, and CP for both groups. Drug-resistance patterns of the resistant strains varied among the 40 types. Among those, prevalent patterns recognized were CP.TC.SM.ABPC, CP.TC.SM.KM.ABPC, TC.SM, SM, and TC.KM in the domestic strains, and TC, CP.TC.SM.ABPC, CP.TC.SM.KM.ABPC, CP.TC.SM.KM.ABPC.ST and TC.KM in the imported strains. The results of the phage-typing test revealed that 31 strains of 52 domestic strains tested, and 13 strains of 46 imported strains tested were definitive type 104 (DT104). Those resistance patterns were CP.TC.SM.ABPC.SU (43 strains) and CP.TC.SM.KM.ABPC.SU (1 strain).

[Incidence of Salmonella serovar Hadar from sporadic cases in Tokyo, and drug resistance of isolates].

A total of 780 Salmonella serovar Hadar (S. Hadar) strains consisting of 601 domestic strains and 179 imported strains isolated in Tokyo, 1980-1998, were examined regarding their incidence and drug-resistance. Domestic strains accounted for 7.2% of all Salmonella (8,359 strains) isolated from domestic cases, and imported strains accounted for 4.4% of all Salmonella (4,083 strains) isolated from imported cases. A drug-resistance test using 9 drugs (CP, TC, SM, KM, ABPC, ST, NA, FOR, and NFLX) showed that 586 strains (97.6%) of the domestic strains and 175 strains (97.8%) of the imported strains were resistant to some of the drugs, excluding NFLX. Drugs with a high resistance rate were TC and SM for both groups. Drug-resistance patterns of the resistant strains varied among the 24 types. Among those, prevalent patterns recognized were TC.SM.KM (231 strains), TC.SM (205 strains), and TC.SM.KM.ABPC (65 strains) in the domestic strains, and TC.SM (135 strains) and TC (13 strains) in the imported strains.

Interconversion of aflatoxin B1 and aflatoxicol by several fungi.

Four fungal strains, namely, Aspergillus niger, Eurotium herbariorum, a Rhizopus sp., and non-aflatoxin (AF)-producing Aspergillus flavus, which could convert AF-B1 to aflatoxicol (AFL), could also reconvert AFL to AF-B1. The interconversion of AF-B1 to AFL and of AFL to AF-B1 was ascertained to occur during proliferation of the fungi. These reactions were distinctly observed in cell-free systems obtained from disrupted mycelia of A. flavus and the Rhizopus sp., but they were not observed in culture filtrates from intact (nondisrupted) mycelia of the same strains. The interconversion activities of AF-B1 and AFL were not observed when the cell-free systems were preheated at 100 degrees C. These findings strongly suggest that the interconversion of AF-B1 and AFL is mediated by intracellular enzymes of A. flavus and the Rhizopus sp. In addition, the isomerization of AFL-A to AFL-B observed in culture medium was also found to occur by the lowering of the culture pH.

Chemically defined medium for high yields of sterigmatocystin.

Isolate of Aspergillus versicolor strain produced 138 micrograms/ml of sterigmatocystin in a complete synthetic medium containing sucrose, salts, 1-phenylalanine, and Ca-pantothenate. The SSP (sucrose salts phenylalanine) medium apparently provided all necessary ingredients for the production of high levels of sterigmatocystin. For optimal sterigmatocystin formation, the amounts of sucrose and 1-phenylalanine were found to be 200 g and 5 g per liter, respectively. When Ca-pantothenate (0.01 g per liter) added, much higher amounts of sterigmatocystin were recovered, whereas CaCl2 addition (0.01%) drastically reduced the yield. The high levels of sterigmatocystin were recovered in the cultures which incubated stationary at 26 to 29 degrees C for over 12 days. Seven strains or isolates tested yielded high levels of sterigmatocystin in the SSP medium, whereas in each other media such as YES medium and rich medium only one isolate yielded highest amount of sterigmatocystin was found.

Fungal contamination and mycotoxin-producing potential of dried beans.

A total of 604 samples of about 7 different types of beans was examined to determine their mycological profiles, and suitability for use as solid substrates for mycotoxin production. All of the samples were collected from bean jam makers in Tokyo by the official food examiners. Genera Penicillium and Aspergillus were predominant, and genus Wallemia was also found commonly in all types of beans. Mycotoxin-producing Aspergillus strains were isolated from 52 samples of beans, approximately 9% of the total. The highest incidence of toxigenic Aspergillus (14.1%) was found in kidney beans. Red beans and peas inoculated with Aspergillus ochraceus were found to produce about 7 to 8 times more toxin than was obtained in a liquid medium. and red beans inoculated with A. versicolor produced more toxin than was obtained in yeast extract sucrose broth. Green peas inoculated with Fusarium graminearum produced about 8 times more T-2 toxin than was obtained in 1% peptone containing Czapek solution under comparable culture conditions.