bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on Group Antigens of Lactobacillus casei
Authors: Shimohashi Hirotaka , Kamiyama Kuniyoshi , Arai Hiroshi
Published: November 2013

Group antigens of Lactobacillus casei were analyzed by immunodiffusion in agar. Antigens were extracted with cold trichloroacetic acid (TCA) from whole cells or cell‐wall preparations. Using cell‐wall extracts as antigens, group B strains were clearly distinguished from group C strains. The antigen specific for group B was designated as antigen 1, and for group C as antigen 2. Group B cells contained either antigen 4 or antigen 5, in addition to antigen 1, in their cell walls, and were divided into two subgroups. TCA extracts of the whole‐cell preparations contained an antigen common to groups B and C. This antigen was called antigen 3. Antigen 3 was not found in the cell‐wall extracts. All species of lactobacilli and some species of other genera were found to contain this antigen in their whole‐cell extracts. Inhibition tests with various sugars showed that L‐rhamnose was the most effective inhibitor for antigens 1 and 5, α‐glucoside for antigen 4, β‐glucoside for antigen 2, and hexosamines for antigen 3.
Slime Production by Pseudomonas aeruginosa
Authors: Murakawa Takeo
Published: November 2013

Chemical properties and compositions of slimes produced by two Pseudomonas aeruginosa strains of different colonial types were investigated. The main component of the slime from strain IFO 3445 was found to be DNA, contaminated with small amounts of protein. On the other hand, the slime from a mucoid‐type strain No. 24 was an alginate‐like substance consisting of mannuronic and glucuronic acids, and contained traces of protein and nucleic acid. Slimes from twenty clinical isolates of P. aeruginosa were investigated for their chemical compositions. Slimes from eighteen strains consisted of DNA, while, two strains of a mucoid‐type produced slimes composed of polyuronic acid.
Adjuvant Activity of Mycobacterial Fractions
Authors: Taniyama Tadayoshi , Watanabe Takeshi , Azuma Ichiro , Yamamura Yuichi
Published: November 2013

The addition of adjuvant (Bacillus Calmette–Guérin‐cell wall skeleton; BCG‐CWS) to the culture medium substantially increased the immune response of mouse spleen cells to immunization with heterologous erythrocytes and hapten–protein conjugate in vitro. Cell walls of mycobacteria, nocardia and corynebacteria and their cell wall constituents were used as adjuvants. In the present case, it was found that cell walls of mycobacteria, nocardia and corynebacteria markedly facilitated primary humoral response of mouse spleen cells to heterologous erythrocytes in vitro. The adjuvant effect of BCG‐CWS was present only in the formation of 19 S antibody in both primary and secondary responses, but not in that of 7 S antibody in vitro. Primary antihapten response of mouse spleen cells against dinitrophenylated keyhole limpet hemocyanin (DNP‐KLH) also succeeded when BCG‐CWS was added to the culture medium, and it was found that BCG‐CWS increased the helper activity of carrier specific helper T cells in vitro where a double chamber system, separated by a cell‐impermeable nucleopore membrane, was used. This result suggested that BCG‐CWS acts on T cells, resulting in the release of soluble factor(s) from T cells capable of exerting an adjuvant effect. Furthermore, mucopeptide moiety of BCG‐CWS retained some adjuvant activity, but other cell wall constituents, such as mycolic acid, arabinose mycolate, and arabinogalactan, did not show any adjuvant effect in vitro. These results strongly imply that mucopeptide moiety of BCG‐CWS plays an important role in the development of adjuvanticity.
Typing of Herpes Simplex Virus Strains of Genital and Nongenital Origins
Authors: Kawana Takashi , Shinkai Kenkichi , Yoshino Kamesaburo
Published: November 2013

Herpes simplex virus strains isolated from genital and nongenital sites were classified into type 1 (HSV‐1) and type 2 (HSV‐2) by endpoint neutralization tests using IgM of rabbits hyperimmunized with either HF (HSV‐1) or UW‐268 (HSV‐2) strain. It was found that about one‐third of the genital isolates belonged to type 1, in contrast to the general concept that HSV‐2 represents genital herpes strains. These HSV‐1 strains, differing from HSV‐2, were mostly isolates from acute herpetic lesions of female patients with constitutional symptoms. On the other hand, all nongenital isolates except one were determined to be HSV‐1. There was no intermediate type equally neutralizable by both types of IgM. A majority of the HSV‐2 strains produced large plaques in chick embryo (CE) cells before passage through avian cells. In contrast, all HSV‐1 strains failed to produce such large CE plaques even after serial passages through avian hosts.
Observations on Nonconverting Phage, c‐n71, Obtained from a Nontoxigenic Strain of Clostridium botulinum Type C
Authors: Oguma Keiji , Iida Hiroo , Inoue Katsuhiro
Published: November 2013

A nontoxigenic mutant (C‐N71) obtained from a toxigenic strain of Clostridium botulinum type C, Stockholm, with nitrosoguanidine treatment was found to be lysogenic by the lysis test. Although the filtrate of a passaged lysate of this nontoxigenic but lysogenic strain, C‐N71, lysed cells of the nontoxigenic strain C‐AO2 equally as well as the converting phage c‐st obtained from the strain C‐Stockholm, it did not convert C‐AO2 to the toxigenic state. The lysis spectrum of this filtrate was the same as that of the c‐st phage. The ability of the filtrate to lyse the indicator cells, C‐AO2, was destroyed neither by trypsin nor DNase but was inactivated by heat treatment at 80 C for 10 min. This suggested that the agent which caused lysis was not boticin but probably a phage. An electron micrograph of the complete phage, c‐n71, which was similar in morphology to that of the c‐st phage was obtained from the filtrate of strain C‐N71. Anti‐c‐n71 phage rabbit serum neutralized both the lytic and the converting activities of the c‐st phage. These findings strongly suggest that the c‐n71 phage is a mutant of the c‐st phage which lacks the gene controlling production of botulinum type C toxin.
Seven‐Year Follow‐Up Study of Rubella Syndrome in Ryukyu with Special Reference to Persistence of Rubella Hemagglutination Inhibition Antibodies
Authors: Ueda Kohji , Nishida Yukiaki , Oshima Kenji , Yoshikawa Hiromi , Ohashi Komei , Nonaka Saneo
Published: November 2013

Rubella hemagglutination inhibition (HI) antibodies in 266 children with rubella syndrome born in 1965 in the Ryukyu Islands and their mothers were followed for seven years. Titers of rubella HI antibody in the mothers declined slowly, while those in the children declined rapidly up to 40 months of age. Thereafter decline of titers became extremely slow and only seven cases (three per cent) became seronegative for rubella HI antibody. Rubella HI antibody titers seemed to have no particular correlation to the severity of clinical manifestations.
Ecology of Enteroviruses in Tropics
Authors: Otatume Sinroku , Addy Patrick A‐K.
Published: November 2013

A continuing surveillance on enterovirus infection in healthy infants was conducted from October 1971 through February 1973 in urban areas in Ghana, West Africa. About 40 infants were visited in every two months for collection of faecal specimens and examined for infection. Enteroviruses were recovered in tissue culture. The overall isolation rate of enteroviruses was approximately 44%, and there was no seasonal difference between rainy and dry seasons. The rate of virus isolation in urban areas was significantly higher than in rural areas. Within the urban areas, however, no difference in the rate of virus isolation was detected between densely populated and sparsely populated areas. The results of virus identification revealed that all three types of Poliovirus, many types of Echovirus and a few Coxsackieviruses were isolated during the course of the study. It was observed that improvement of sanitary facilities decreased the frequency of virus infection among infants, but the condition of water supply did not influence the virus infection rate. Neither the age of infants nor the size of siblings showed any relation to the virus isolation rate. It was suggested from the results that many types of enteroviruses have been circulating continuously in the tropical urban community throughout the year.
Amoxycillin and Ampicillin
Authors: Miller M.A. , Kuemmerle N.B. , Gentile G.
Published: November 2013

In vitro antibacterial activities of ampicillin and amoxycillin were compared against pigmented and non‐pigmented strains of Serratia marcescens. Ampicillin appeared more effective than amoxycillin; three‐fourths of all strains consistently exhibited an ampicillin minimum inhibitory concentration (MIC) of at least one tube less than that recorded for amoxycillin. Complete cross resistance was not observed as has previously been inferred. Further, greater bactericidal activity was demonstrated with ampicillin; minimum bactericidal concentrations (MBC) were either the same as or one tube greater than the MIC. MBCs for amoxycillin, however, were significantly higher; often four to five times greater than the MIC. Ampicillin exhibited greater bactericidal activity as inferred from differences observed in the biological lesions induced, as recorded through observations by scanning electron microscopy (SEM). Spheroplasts were the predominant morphological alteration induced by ampicillin. In contrast, only filament formation, which demonstrated a degree of reversibility, was induced by amoxycillin.
Isolation and Properties of Escherichia coli Mutants Which Are Nonpermissive for the Growth of Phage Lambda
Authors: Takahashi Seishi , Hidaka Saburo , Matsubara Kenichi
Published: November 2013

By selecting survivors of λ phage infection, mutants of Escherichia coli K12 that block reproduction cycle of the phage have been isolated. Fourteen of these phage‐tolerant mutants (lam mutants) were chosen and characterized biochemically and genetically. It was shown that these mutants were tolerant to infection by all the lambdoid phages, except for few cases, but they were susceptible to infection by a non‐lambdoid temperate phage (φ299), P1 or T phages. The mutants can be divided into at least three groups: (1) A mutant (lam 16) strain that seems to block normal penetration of phage DNA: (2) Three mutant (lam 64, lam 67 and lam 71) strains that block an “early” step(s) of phage growth, including phage DNA synthesis: (3) Six mutant (lam 24, lam 25, lam 26, lam 27, lam 646 and lam 6) strains that block normal functioning of the gene E products and produce unusual head structures. Some lambdoid phages and λ mutants that overcome the interference by the lam mutations have been obtained, and were used as tools for characterizing the host mutations. Two (lam 12 and lam 13) mutant strains and one (lam 1) mutant were inferred as affecting the expression of “late” genes, and early gene, respectively, by this test.
Mouse Hepatitis Virus (MHV‐2)
Authors: Hirano Norio , Fujiwara Kôsaku , Matumoto Minoru
Published: November 2013

Various factor sinfluencing the plaque formation of mouse hepatitis virus (MHV‐2) in DBT cell monolayers were studied and a practical method for plaque assay was developed. Infected DBT cells yielded high‐titered virus and were a satisfactory source of complement‐fixing viral antigen. The predominant cytopathic effect of MHV‐2 in DBT cells was cell rounding and detachment, but no syncytial formation was observed. Fluorescent antibody staining revealed specific fluorescence only in the cytoplasm of infected DBT cells. In one‐step growth experiment, newly formed virus was first recognized within 4‐hr postinfection and showed subsequently a rapid exponential increase. Release of newly formed virus from the cell was rapid, and a continuous release lasted for a certain period of time. The average per‐cell yield of active virus was estimated to be about 6–7 × 102 plaque‐forming units.
Difference in Response to Mouse Hepatitis Virus among Susceptible Mouse Strains
Authors: Taguchi Fumihiro , Hirano Norio , Kiuchi Yoshihiro , Fujiwara Kôsaku
Published: November 2013

After intraperitoneal inoculation with a high‐virulent mouse hepatitis virus (MHV) a significant difference was seen in survival time between DDD and CDF1 (BALB/c × DDD) mice, while 50% lethal doses were not significantly different. With 3 × 103 PFU of the virus CDF1 and DDD mice died in 45 and 120 hr, respectively, on the average. This difference of susceptibility between DDD and CDF1 mice was first demonstrable at the age of 1 week and was more pronounced at the age of 4 weeks but showed no dependence on the sex. Virus titers ran 2 to 3 log higher in the liver and blood of CDF1 than in those of DDD mice, while being only 1 log higher in the spleen. At an early stage of infection viral antigen was demonstrable by immunofluorescence in sinusoidal lining cells of the liver more prominently in CDF1 than in DDD mice. Interferon production occurring in parallel with virus growth was significantly higher in CDF1 than in DDD mice. In DDD mice, liver lesions were rather focal with some accumulation of round cells, while they were confluent with poor cellular response in CDF1 mice. Viral growth in cultured peritoneal macrophages from CDF1 mice was 1 log higher than in those from DDD mice. The results suggest that the divergence in response to MHV among susceptible mice greatly depends upon the susceptibility of macrophages and reticuloendothelial cells which constitute primary targets of the virus.
Temperate Coliphage HK022: Clear Plaque Mutants and Preliminary Vegetative Map
Authors: Dhillon T.S. , Dhillon Elvera K.S.
Published: November 2013

Wild type phage HK022 was mutagenized by N‐methyl‐N′‐nitro‐N‐nitrosoguanidine to induce clear plaque mutants. A total of 225 clear plaque mutants were isolated and 198 of these were assignable to one or the other of the two complementation groups of the corresponding cistrons which have been designated as cI and cII, respectively. Approximately 25% of the c mutants were found to be temperature‐sensitive (cts); producing turbid plaques at 32 C and clear plaques at 38 C and above. From complementation tests involving cI and cII mutants, bacteria lysogenic for cII prophage were frequently obtained. Double lysogens harboring a cI and a cII prophage were infrequently found and single lysogens harboring only a cI prophage have not been recovered. Bacterial lysogens harboring a prophage carrying a cts mutation in the cI cistron were readily obtainable. However, such lysogens show a lethal phenotype at 40 C and above, although they appear to be fully viable at 32 C. It is shown that by incubation of lysogens harboring a cts mutant of the cI cistron at 42 C, it is possible to isolate cryptic lysogens which are non‐immune but harbor at least one of the phage sus+ alleles. Genetic data involving cI, cII, and two complementing sus mutants of essential genes are presented. From these data the following vegetative map is deduced: sus4–cII‐cI‐sus3.
Distribution of Indigenous Lactobacilli in the Digestive Tract of Conventional and Gnotobiotic Rats
Authors: Watanabe Tsugio , Morotomi Masami , Suegara Nobuo , Kawai Yasuo , Mutai Masahiko
Published: November 2013

Distribution of indigenous lactobacilli in the gastrointestinal tracts of rats was investigated at the species level. The indigenous lactobacilli isolated from conventional rats were divided into three groups, Lactobacillus acidophilus and its related strains, L. fermentum, and L. murini. Localization of the Lactobacillus groups in the gastrointestinal tracts could be distinguished clearly based on arabinose and glucose fermentation reaction of isolates from each part of the gastrointestinal tract. Group I (L. acidophilus and the related strains) and Group II (L. fermentum) were the major populations of lactobacilli on the walls of the non‐glandular part and in the contents of the stomachs of both conventional and gnotobiotic rats. Group I predominated in all the parts of the digestive tract of conventional rats, whereas Group II was in the minority in the lower part of the gastrointestinal tracts of both groups of rats. Group III (L. murini) was the predominant population of Lactobacillus in the lower part of small intestine of conventional rats and in all parts of the gastrointestinal tracts of gnotobiotic rats except for the wall of the non‐glandular part of the stomach.
Thermolabile Repression of Cephalosporinase Synthesis in Citrobacter freundii
Authors: Sawai Tetsuo , Nakajima Sachio , Morohoshi Toshiro , Yamagishi Saburo
Published: November 2013

An unusual regulatory system of cephalosporinase synthesis in Citrobacter freundii has been found. When the bacteria are grown at 20 C, the cephalosporinase is synthesized as a typical inducible enzyme and benzylpenicillin acts as an effective inducer. The enzyme, however, is synthesized in the absence of the inducer at growth temperatures above 25 C. When the growth temperature is shifted from 20 C to 37 C, the induction of enzyme synthesis is observed after about one half of the organism doubling time, but it does not occur in the presence of chloramphenicol. The reverse control mutants, the enzyme constitutive synthesis of which is markedly depressed by benzylpenicillin, were isolated from the C. freundii wild strain. The possibility that the enzyme synthesis is governed by a regulatory system analogous to the its mutant of the lac operon in Escherichia coli was suggested.
Dental Caries Induction in Experimental Animals by Clinical Strains of Streptococcus mutans Isolated from Japanese Children
Authors: Hamada Shigeyuki , Ooshima Takashi , Torii Mitsuo , Imanishi Hideaki , Masuda Norio , Sobue Shizuo , Kotani Shozo
Published: November 2013

Oral implantation and the cariogenic activity of clinical strains of Streptococcus mutans which had been isolated from Japanese children and labeled with streptomycin‐resistance were examined in specific pathogen‐free Sprague‐Dawley rats. All the seven strains tested were easily implanted and persisted during the experimental period. Extensive carious lesions were produced in rats inoculated with clinical strains of S. mutans belonging to serotypes c, d, e, and f, and maintained on caries‐inducing diet #2000. Noninfected rats did not develop dental caries when fed diet #2000. Type d S. mutans preferentially induced smooth surface caries in the rats. Strains of other serotypes primarily developed caries of pit and fissure origin. Caries also developed in rats inoculated with reference S. mutans strains BH‐TR and FAIR (type b) that had been maintained in the laboratories for many years. However, the cariogenicity of the laboratory strains was found to have decreased markedly. All three S. sanguis strains could be implanted, but only one strain induced definite fissure caries. Two S. salivarius strains could not be implanted well in the rats and therefore they were not cariogenic. Four different species of lactobacilli also failed to induce dental caries in rats subjected to similar caries test regimen on diet #2000. S. mutans strain MT6R (type c) also induce caries in golden hamsters and ICR mice, but of variable degrees.
In Vitro Immune Response of Human Peripheral Lymphocytes
Authors: Kuritani Taro , Hirano Toshio , Kishimoto Tadamitsu , Yamamura Yuichi
Published: November 2013

Con A‐stimulation of human peripheral T lymphocytes induced both suppressor and helper T cells. Con A‐generated suppressor T cells inhibited PWM‐indueed IgG and IgM production in PBL. Lower concentrations of Con A (0.5 μg/ml) or shorter incubation periods (6 to 24 hr) induced mainly helper T cells, while higher concentrations of Con A (10 μg/ml) or longer incubation periods (at least 48 hr) induced suppressor T cells. Con A‐generated suppressor T cells were sensitive to mitomycin treatment and exerted their suppressor function on the early phase of differentiation and/or proliferation of B cells but not on the final differentiation of B cells to Ig‐producing cells. The identity of the MHC was not required for the expression of suppressor function. Suppressor T cells competed with helper T cells in PWM‐induced Ig‐production of PBL. This experimental system can be applied to estimate the regulatory function of T cells in several disease states.
Polyclonal B Cell Activation by Cell Wall Preparations of Gram‐Positive Bacteria
Authors: Saito‐Taki Tatsuo , Tanabe Masao J. , Mochizuki Hitoshi , Matsumoto Takao , Nakano Masayasu , Takada Haruhiko , Tsujimoto Masachika , Kotani Shozo , Kusumoto Shoichi , Shiba Tetsuo , Yokogawa Kanae , Kawata Shigeo
Published: November 2013

The effects of polyclonal B cell activation (PBA) of cell walls and their cell wall fractions obtained from several kinds of gram‐positive bacteria were studied using the anti‐sheep red blood cell (SRBC) or anti‐trinitrophenylated (TNP) SRBC plaque forming cell (PFC) responses of cultured spleen cells from Balb/c, athymic nu/nu, their littermates (nu/+), C3H/He (LPS‐responder), C3H/HeJ (LPS‐non‐responder), (CBA/N × Balb/c) F1 male with an X‐linked defect in B cell function and the F1 female mice. The cell walls of Staphylococcus epidermidis (ATCC 155), Lactobacillus plantarum (ATCC 8014), Micrococcus lysodeikticus (NCTC 2665), Mycobacterium rhodochrous (ATCC 184), Streptomyces gardneri (ATCC 23911) and Nocardia corynebacteriodes (ATCC 14898) had the ability to induce polyclonal B cell responses in the spleen cells of Balb/c, nu/nu, nu/+, C3H/He and C3H/HeJ mice. The cell wall fractions prepared by enzymatic digestion from the cell walls of S. epidermidis, S. gardneri or N. corynebacteriodes were also capable of inducing polyclonal B cell responses. The responses of spleen cells from (CBA/N × Balb/c) F1 male mice to these active preparations, except the cell walls of M. rhodochrous, were much lower than those of the F1 female mice. These findings indicate that the majority of the cell wall preparations lacks PBA ability for spleen cells with the CBA/N defect, except for the cell walls of M. rhodochrous which possess this ability. The PBA‐ability of synthetic peptidoglycan, muramyl dipeptide (N‐acetylmuramyl‐L‐alanyl‐D‐isoglutamine, MDP), was also examined, and a similar activity was observed in MDP.
An Outbreak of Acute Enteritis Due to Campylobacter fetus Subspecies jejuni at a Nursery School in Tokyo
Authors: Itoh Takeshi , Saito Kahiko , Maruyama Tsutom , Sakai Senzo , Ohashi Makoto , Oka Aiko
Published: November 2013

An outbreak of acute enteritis due to Campylobacter fetus subspecies jejuni involving a total of 35 out of 74 children occurred at a nursery school in Tokyo in January 1979 and lasted for 7 days. It was the first case of a community outbreak of the disease to be recognized in Japan.

The major symptoms observed in the patients consisted of diarrhea (88%), fever (82%), abdominal pain (39%), and vomiting (6.1%). The rate of isolation of the organism from the patients was 39%. Sera of four patients showed elevated agglutinin titers against the organism ranging from 1: 80 to 1: 320.

Although it is suggested that the outbreak was caused by a communal lunch or between‐meal snacks prepared by and provided at the nursery school, the incriminated food, source and route of contamination could not be pinpointed.
Establishment of Autoantibody‐Producing Cell Lines from Peripheral Blood Lymphocytes of Patients with Systemic Lupus Erythematosus
Authors: Hirano Toshio , Teraoka Osamu , Teranishi Tsuyoshi , Tsityuguchi Izuo , Tohda Hiroko , Oikawa Atsushi
Published: November 2013

Autoantibody‐producing B cell lines were established from peripheral blood lymphocytes of patients with systemic lupus erythematosus. Peripheral blood lymphocytes obtained from five of seven patients were successfully transformed by Epstein‐Barr virus. Two of four established B lymphoblastoid cell lines examined in this study produced anti‐nuclear factor antibodies and one of them produced anti‐single‐stranded DNA and anti‐double‐stranded DNA antibodies. These results indicate that B cell clones committed to self antigens are transformed by Epstein‐Barr virus and continue to produce autoantibodies. In order to establish a monoclonal autoantibody‐producing B cell line, the cells were cloned by a limiting dilution method. The data suggest that it is possible to establish a monoclonal autoantibody‐producing B cell line by the combination of transformation of B cells by Epstein‐Barr virus and extensive cloning.
Amino Acid Requirements for the Growth of Aedes albopictus Clone C6/36 Cells and for the Production of Dengue and Chikungunya Viruses in the Infected Cells
Authors: Sasao Fuyoko , Igarashi Akira , Fukai Konosuke
Published: November 2013

Amino acid requirements for the growth of Aedes albopictus, clone C6/36, cells and for the production of dengue (DEN) and Chikungunya (CHIK) viruses were examined by growing the cells or the viruses in media which were deprived of one of the 20 amino acids. Cell growth was markedly inhibited when cystine was omitted from the medium, and to a lesser extent by arginine deprivation. On the other hand, omission of alanine, asparagine, aspartic acid, and glutamic acid at the same time did not affect cell growth. Marked accumulation of alanine was observed in the medium when the cells were grown for 8 days in complete medium, with concomitant depletion of aspartic acid and glutamic acid. The production of CHIK virus was inhibited markedly by omission of cystine from the medium after virus infection, while the production of DEN viruses was more affected by glycine deprivation, although cystine deprivation also inhibited virus production to a lesser extent. On the other hand, production of CHIK and DEN viruses was not affected when alanine, asparagine, aspartic acid, and glutamic acid were omitted from the medium at the same time.
Microbial Adjuvant and Autoimmunity
Authors: Yamaki Kenichi , Ohta Michio , Nakashima Izumi , Noda Aiji , Asai Junpei , Kato Nobuo
Published: November 2013

Definite lesions in the exocrine pancreas were produced when SMA mice were immunized eight times at intervals of 30 days with a mixture of extract of pooled pancreas from syngeneic mice and the capsular polysaccharide of Klebsiella pneumoniae type 1 Kasuya strain (CPS‐K), whereas no pancreatic lesions were produced in mice given CPS‐K alone or pancreatic extract alone. The typical histological changes were characterized by infiltration with lymphocytes, plasma cells, and other mononuclear cells, degeneration and lysis of the acinar cells, destruction of the lobular architecture, and replacement by fatty tissue and fibrous connective tissue. The endocrine islets were well preserved. No specific histological changes were produced in the organs other than the pancreas in these mice. Most of mice immunized with pancreatic extract mixed with CPS‐K produced serum precipitins to syngeneic pancreatic antigens. However, severe pancreatic lesions were also produced in mice showing no definite precipitin production.
Deconjugation of Bile Salts by Bacteroides and Clostridium
Authors: Masuda Noriyuki
Published: November 2013

Deconjugation of bile salts by four strains of Bacteroides and four strains of Clostridium was studied by use of resting cells and cell‐free culture supernatants. Bacteroides strains yielded active cells but showed relatively low bile salt hydrolase (BSH) activity in the culture supernatants while the reverse was the case for the spore‐forming clostridial strains.

BSH was formed constitutively and was oxygen insensitive. The optimum pH was between 4.5 and 5.0. Marked substrate specificity was found in two strains, one Clostridium and one Bacteroides, which showed restricted activity against taurine conjugates. Bacteroides in general attacked the taurine conjugates of di‐hydroxy bile acids more readily than the trihydroxy taurine conjugates.

Deconjugated bile acid moieties were further modified by some resting cells, depending on the bacterial strain while no enzymatic activity other than that of BSH was found in the culture supernatants. Cells of B. fragilis 2536 performed 7α‐dehydrogenation when the pH of the medium allowed the reaction, and this oxidative process was markedly enhanced in the presence of an abundant supply of oxygen as a terminal electron acceptor. C. perfringens PB 6K produced the 3‐keto product in addition to the 3β‐hydroxy derivative of the liberated bile acids and the formation of the latter derivative seemed to take place without preliminary deconjugation.
A Comparative Study of the Sugar Composition of O‐antigenic Lipopolysaccharides Isolated from Vibrio alginolyticus and Vibrio parahaemolyticus
Authors: Hisatsune Kazuhito , Kiuye Akira , Kondo Seiichi
Published: November 2013

A comparative study of the sugar composition of O‐antigenic lipopolysaccharides (LPS) isolated from Vibrio alginolyticus and those from V. parahaemolyticus was carried out. 3‐Deoxy‐d‐mannooctulosonic acid, 2‐keto‐3‐deoxy octonate (KDO), a regular sugar constituent of gram‐negative bacterial LPS, was totally absent from LPS of all V. alginolyticus strains examined as it was from those of V. parahaemolyticus. Furthermore, a KDO‐like thiobarbituric acid test‐positive substance, identical with that of either V. parahaemolyticus 07 or 012, was also found in LPS from three strains, 505–78, 905–78, and 1013–79 (designated tentatively as group I), out of the five strains of V. alginolyticus tested. LPS from the members of group I contained, as component sugars, glucose, galactose, l‐glycero‐d‐manno‐heptose, glucosamine, galactosamine, the KDO‐like substance, and an unidentified amino sugar P1. Thus, LPS of the members of group I possessed a similar sugar composition which is similar to that of LPS from either V. parahaemolyticus 07 or 012. LPS of strain 1027–79, one of the other two strains (designated tentatively as gorup II), contained as component sugars, glucose, l‐glycero‐d‐mannoheptose, glucosamine, galactosamine, and the other unidentified amino sugar P2, while LPS of strain 53–79, the other member of group II, contained galactose as an additional component. The results indicate that LPS of strain 1027–79 has a sugar composition similar to that of V. parahaemolyticus 09 LPS.
Lipid Peroxide Formation and Membrane Damage in Endotoxin‐Poisoned Mice
Authors: Sakaguchi Shuhei , Kanda Norio , Hsu Cheng‐Chin , Sakaguchi Osamu
Published: November 2013

Lipid peroxide formation and plasma membrane damage in mouse liver following the administration of Salmonella endotoxin were examined. The liver lipoperoxide level was markedly elevated in animals given endotoxin compared with that in the controls, and returned to its normal range after 2 days. On the other hand, superoxide dismutase activity was decreased by 18–48 hr after endotoxin injection, thereafter tending to increase. Glutathione reductase and glutathione peroxidase activities declined in the liver 18 hr after the injection. The endotoxin resulted in much lower lipoperoxide formation in the livers of tolerant mice than in those of the poisoned mice. The lipoperoxide level in endotoxin‐poisoned mice after the administration of α‐tocopherol was lower than that in the controls, and α‐tocopherol administration prevented completely the membrane protein damage that arose from endotoxin challenge. After glutathione administration the membranes of the poisoned mice also returned to almost the normal disk electrophoretic profile. These results suggest that lipid peroxide formation in the liver plasma membrane caused by free radicals might occur in a tissue ischemic state in endotoxicosis.
Isolation of Clostridium difficile from the Feces and the Antibody in Sera of Young and Elderly Adults
Authors: Nakamura Shinichi , Mikawa Masato , Nakashio Satoshi , Takabatake Manabu , Okado Imiko , Yamakawa Kiyotaka , Serikawa Toshihiko , Okumura Seiichi , Nishida Shoki
Published: November 2013

Attempts were made to isolate Clostridium difficile from a total of 431 fecal specimens from 149 young and 213 elderly healthy adults, and 69 elderly adults with cerebrovascular disease but no gastrointestinal disease. C. difficile was isolated from 49 specimens, and the frequency of isolation was 15.4% in healthy young adults, 7.0% in healthy elderly adults, and 15.9% in elderly adults with cerebrovascular disease. Thirty‐four (about 70%) of the 49 C. difficile strains isolated produced cytotoxin which was neutralized by Clostridium sordellii antitoxin in vitro; in both young and elderly adults approximately 30% of the C. difficile isolates were nontoxigenic. The mean concentration of C. difficile in feces was 104.1/g in young adults and 104.6/g in elderly adults, with a range of 102.0 to 106.9/g. Antibody against C. difficile toxin was found in most of the sera obtained from young adults carrying toxigenic C. difficile, but not in sera of elderly adults, no matter how abundant was toxigenic C. difficile in the feces.
Pharmacological Effect of Beta Toxin of Clostridium perfringens Type C on Rats
Authors: Sakurai Jun , Fujii Yoshio , Matsuura Miho , Endo Koichi
Published: November 2013

The biological effect of purified beta toxin of Clostridium perfringens type C in vivo was investigated. After intravenous injection of the purified beta toxin into rats, a rise in blood pressure and a simultaneous fall in heart rate were observed. After the blood pressure reached a maximum, the heart rate recovered gradually, and electrocardiographic and respiratory changes began. The rise in blood pressure induced by beta toxin tended to be proportional to the amount of toxin. The latent period between the injection of toxin and the onset of the increase, and also the time between the injection and the maximum pressure induced by the toxin decreased with increasing concentration of the toxin. A good correlation was found between the factor producing the rise in blood pressure and beta toxin. Alpha adrenergic and ganglionic blocking agents reduced blood pressure levels elevated by beta toxin. The data suggest that the toxin causes a release of catecholamines, and that the increase in blood pressure was induced by released catecholamines.
Behavior of Mycoplasma hominis in a Human Diploid Cell Culture System
Authors: Sasaki Tsuguo , Shintani Miharu , Yamada Kenichiro , Okumura Hideo , Kihara Kojo
Published: November 2013

The behavior of Mycoplasma hominis in normal human embryonic lung fibroblast (HAIN‐55) cell cultures was investigated. Multiplication patterns of cell‐associated mycoplasmas and of extracellular mycoplasmas in the HAIN‐55 cultures depended upon the size of the inoculum. This relationship did not vary with the number of days the cells had been cultured, nor with the number of HAIN‐55 cell passages. The maximum mycoplasmal growth was obtained with inoculum sizes of 105 to 106 colony‐forming units (CFU)/ml. The recovery of mycoplasmas decreased rapidly with inoculum size beyond 107 CFU/ml, and growth of the HAIN‐55 cells was inhibited. Growth of the cells was also inhibited by the addition of the cytoplasmic fraction of Mycoplasma hominis.
Murine Defense Mechanism against Candida albicans Infection: I. Collaboration of Cell‐Mediated and Humoral Immunities in Protection against Systemic C. albicans Infection
Authors: Kagaya Keiko , Shinoda Takako , Fukazawa Yoshimura
Published: November 2013

Mice immunized with viable C. albicans cells demonstrated a high incidence of cell‐mediated and a low incidence of humoral immune response. There was good agreement between the final survival rate of C. albicans infected mice and the rate of simultaneous cell‐mediated and humoral immune response acquisition. Immunized mice with positive delayed hypersensitivity (DTH) against C. albicans crude antigen showed significant protection against intravenous challenge with C. albicans. Furthermore, the transfer of immunoglobulins from rabbit anti‐C. albicans serum to DTH‐positive mice enhanced protection, while it did not protect control mice against a subsequent challenge with C. albicans. These results suggest that cell‐mediated immunity plays a major role and humoral immunity a side role in the defense mechanism(s) of C. albicans infected mice.
Antigenic Analysis of Polioviruses Isolated from a Child with Agammaglobulinemia and Paralytic Poliomyelitis after Sabin Vaccine Administration
Authors: Hara Minoru , Saito Yoshiko , Komatsu Toshihiko , Kodama Hideo , Abo Wataru , Chiba Shunzo , Nakao Toru
Published: November 2013

A 3‐year‐old boy with agammaglobulinemia developed paralytic poliomyelitis on day 553 after being fed poliovaccine. Non‐vaccine‐like type 2 polioviruses were isolated from 22 stools obtained within 684 days after the onset of illness. Antigenic variations were observed among these viruses. The non‐vaccine‐like virus isolated 1 week after the onset of paralysis differed in virulence from the Sabin type 2 vaccine strain in the neurovirulence test in monkeys, and did not have the same antigenic character as the wild virulent strains. Another virus isolated on day 348 before the onset of illness was also classified as non‐vaccine‐like. However, the Sabin type 2 strain was shown to be homologous with this strain by the McBride test. Some Sabin‐like particles were found in this stock virus. We may conclude that the non‐vaccine‐like virus isolates were derived from Sabin vaccine by antigenic variation that occurred during long‐term multiplication in the intestinal tract.
The Effects of Temperature and pH on the Growth of Eight Enteric and Nine Glucose Non‐Fermenting Species of Gram‐Negative Rods
Authors: Tsuji Akiyoshi , Kaneko Yasuko , Takahashi Kuniko , Ogawa Masatoshi , Goto Sachiko
Published: November 2013

We studied the heat resistance and the range of growth temperature o gram‐negative rods to find one of the bacterial factors governing their infectivity in exogenous and endogenous infections in predisposed patients.

Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Pseudomonas aeruginosa, and Acinetobacter calcoaceticus grew equally well at 25, 30, 37, and 42 C. Among other sugar non‐fermenting gram‐negative rods, six species showed suppressed growth at either or both ends of the incubation temperature range. All the bacterial species tested were killed within 30 min at 60 or 70 C. At 10 C, none of the bacterial strains multiplied, but all survived for 6 hr.

Of 17 bacterial species tested, E. coli had the widest range of growth temperature (18–47 C), and also the shortest time necessary for growth to a certain population. Among the sugar non‐fermenting rods, A. calcoaceticus had the widest range of growth temperature (20–45 C) and also multiplied rapidly. Pseudomonas strains exhibited slower growth at all temperatures and also had a narrower range of growth temperature than Enterobacteriaceae. Among Pseudomonas species, P. aeruginosa had the widest range of growth temperature (25–42 C) and also showed rapid growth. Pseudomonas cepacia, Achromobacter xylosoxidans, and Alcaligenes faecalis had a narrow range of growth temperature (28–37 C), and Pseudomonas fluorescens, Flavobacterium meningosepticum, and Moraxella grew most rapidly at 30 C.

The above results are correlated fairly well with the incidence of clinical cases of infection. The growth attitude of a species of bacteria in response to temperature was considered to be one of the factors affecting the establishment of infection.
Purine Metabolic Enzymes in Lymphocytes: I. Adenosine Deaminase and Purine Nucleoside Phosphorylase Activities in Mouse Lymphocyte Subpopulations
Authors: Kurashige Satonori , Akuzawa Yuki , Yoshida Toshiharu , Teshima Chisato , Mitsuhashi Susumu
Published: November 2013

The distribution of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) activities in lymphoid organs and lymphocyte subpopulations in mice, and the effect of phytohemagglutinin P (PHA‐P) and concanavalin A (Con A) on the enzyme activities were studied. ADA activity was distributed equally in cells from all organs used and no mouse strain differences were observed. In contrast, PNP activity varied with the mouse strain, being highest in C57BL/6 mice and lowest in BALB/c mice, and with the organ in ICR mice, being high in peripheral blood lymphocytes and spleen lymphocytes, low in mesenteric lymph node cells and absent or very weak in thymus cells. T and B lymphocytes were prepared from spleens of ICR mice. High ADA activity was found in both T and B lymphocytes, whereas PNP activity in the T lymphocytes was about one‐third of that in the B lymphocytes. PNP activity in thymus cells was increased to the normal level of T lymphocytes in the spleens by cultivation without stimulant. The development of PNP activity in thymus cells was partially inhibited by Con A but was not affected by PHA‐P. ADA activity in thymus cells was enhanced by in vitro stimulation with PHA‐P but not with Con A. In contrast, in spleen lymphocytes the development of ADA activity was enhanced by stimulation with PHA‐P and Con A, and that of PNP activity was enhanced by PHA‐P but not by Con A.
Immunochemical Characterization of Candida albicans Cell Wall Antigens: Specific Determinant of Candida albicans Serotype A Mannan
Authors: Suzuki Motofumi , Fukazawa Yoshimura
Published: November 2013

We investigated the chemical structure of the specific determinant in the mannan of Candida albicans M‐1012 (serotype A) strain. Acetolysis of the mannan, obtained by alkali extraction and purified as the copper complex, gave mannose and six oligosaccharides (from di‐ to hexasaccharide) and a small amount of a heptasaccharide. We examined the inhibition by these oligosaccharides up to hexaose of the precipitin reaction between anti‐factor 6 serum specific for serotype A and homologous mannan, and found that the mannohexaose was the most effective inhibitor. These, and results obtained by proton magnetic resonance (PMR) spectroscopy, methylation analysis, and other structural studies, suggest that the main component of this hexaose consists of one terminal α(1–3) linkage in addition to four α(1–2) linkages, and that this α(1–3)‐containing mannohexaose may be responsible for the specificity of antigenic factor 6. Further results obtained by analyses of polarimetry, PMR spectroscopy, and chromium trioxide oxidation‐methylation of C. albicans M‐1012 mannan and its acid‐degraded mannans suggest that the mannan has a β‐linkage in addition to α‐linkages, and that the mode of the β‐linkage is mainly (1–6) linkage. Further evidence obtained by Smith degradation‐methylation analysis and by quantitative precipitin reactions of intact and acid‐degraded mannan suggests that the antigenic determinant of antigenic factor 6 may be bound, via the β(1–6) linkage, to C‐6 of mannose residues involved in oligosaccharide side chains of serotype A mannan.
Histopathological Studies of Reactions in Mice Injected with Aluminum‐Adsorbed Tetanus Toxoid
Authors: Goto Norihisa , Akama Kiyoto
Published: November 2013

Histopathological observations of changes at the injection site in mice were made for 20 weeks after a single intramuscular (im) injection of aluminum hydroxide‐adsorbed tetanus toxoid (Alum‐TT), plain tetanus toxoid (TT), or aluminum adjuvant (Alum alone). Marked injury to muscle fibers and infiltration of neutrophils around the aluminum remnants were observed, and some microabscesses were occasionally formed, after im injection of Alum‐TT. Some mature alum‐granulomas were seen in the Alum‐TT group, while the Alum alone group had only a few immature alum‐granulomas. The TT group showed only slight injury to muscle fibers and acute inflammation at an early stage. In the Alum‐TT group, well‐developed lymphoid tissues and granulomatous lesions were still observable even 20 weeks after the injection; however, these changes had already diminished in the Alum alone group by that time. These findings could serve as good models for undesirable local reactions to aluminum‐adjuvanted vaccines.
Immune Response to Toxoplasma gondii
Authors: Yano Akihiko , Yui Katsuyuki , Yamamoto Masuji , Aosai Fumie , Furuta Seiichi , Kojima Soumei
Published: November 2013

Peripheral blood leukocytes (PBL) from patients with toxoplasmosis were shown to be highly responsive to in vitro stimulation with Toxoplasma gondii extract as measured by incorporation of [3H]methylated thymidine. Analysis of Toxoplasma‐specific proliferative cells in PBL by using monoclonal antibodies specific for human T cell subsets revealed that the Toxoplasma‐specific proliferation response of PBL from the patients was mediated by Leu 1, Leu 3a positive cells, that is, helper/inducer T cells. Tests for the Toxoplasma‐specific proliferation response may provide a readily available method for the diagnosis of congenital toxoplasmosis, especially during the newborn period.
Effect of High Salt Treatment on Influenza B Viral Protein Synthesis in MDCK Cells
Authors: Tsurumi Tatsuya , Aoki Hiizu , Nishiyama Yukihiro , Shibata Motohiro , Maeno Koichiro , Seo Hisao
Published: November 2013

Based on the information that high salt inhibits the initiation of cellular mRNA translation which depends on the function of the 5′‐terminal structure of mRNA, we compared the effect of high salt on translation of host cellular mRNAs and influenza viral mRNAs, both of which are of 5′‐terminal structure. Brief exposure of influenza B virus‐infected MDCK cells to high salt medium resulted in a dose‐dependent inhibition of viral polypeptide synthesis as well as of cellular polypeptide synthesis, but it had less effect on synthesis of viral polypeptides, particularly nonstructural protein (NS). Under these conditions the Na+ content of the infected cells was significantly increased. A similar salt effect on in vitro translation of viral and cellular mRNAs extracted from infected cells was also observed. There was no significant difference in sensitivity to hypertonic block of in vivo translation of influenza viral mRNAs and vesicular stomatitis virus mRNAs, the latter of which possess a virus‐directed structure at the 5′‐terminus.
Growth of the Surface of Corynebacterium diphtheriae
Authors: Umeda Akiko , Amako Kazunobu
Published: November 2013

Surface structure and growth of the surface of Corynebacterium diphtheriae mitis strain were investigated by scanning electron microscopy and the immunofluorescence technique. The surface of the cell revealed by the scanning electron microscope showed a few elevated circular zones which encompassed the cell. The cell diameter increased at this zone and this gave the club‐shaped appearance to this species. The cell surface labeled with specific antibodies against the whole bacterial cell and tagged with ferritin remained at a constant length during cell division cycles and the new cell surface emerged from the polar ends of the cell. This new wall surface was completely devoid of the ferritin particles indicating that the cell wall component on the old preexistent wall was completely conserved. A similar finding was obtained by immunofluorescence microscopy. C. diphtheriae, unlike Bacillus spp., showed apical growth as has been observed in fungal cells.
Effects of Bordetella pertusiss Components on IgE and IgG1 Responses
Authors: Sekiya Kachiko
Published: November 2013

The effect of dermonecrotic toxin (DNT), fimbrial hemagglutinin (FHA), K‐agglutinogen, lipopolysaccharide (LPS), and pertussigen from Bordetella pertussis on the production of IgE and IgG1 antibodies to hen egg albumin (Ea) was investigated in C57BL/6 mice. The IgE antibody contents were determined by passive cutaneous anaphylaxis (PCA) in the skin of Lewis rats, while the IgG1 antibody contents were determined by PCA reactions on the skin of mice using sera that had been heated for 3 hr at 56 C to destroy the IgE antibodies. Among the B. pertussis components tested, pertussigen was the most effective adjuvant for increasing the IgE and IgG1 antibodies to Ea. LPS also moderately increased both types of antibodies, and FHA slightly increased the IgG1 titers. When LPS was given 5 days before Ea, it suppressed both IgE and IgG1 titers while FHA had only slight adjuvant action on both type of antibodies.

When each of the components was tested for its ability to modify the adjuvant action of pertussigen, it was found that only DNT interfered significantly with the adjuvanticity of pertussigen when given on the day of immunization with Ea. When the components were given 5 days before Ea, DNT produced significant suppression of only the IgG1 response. LPS, FHA, and K‐agglutinogen did not significantly affect the adjuvant action of pertussigen.