bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of N‐Acetylchitohexaose against Candida albicans Infection of Tumor‐Bearing Mice
Authors: Kobayashi Makiko , Watanabe Toshihiko , Suzuki Shigeo , Suzuki Masuko
Published: November 2013

A water‐soluble oligosaccharide, N‐acetylchitohexaose (NACOS‐6), was able to enhance the protective effect against Candida albicans infection in mice during the early phase of tumor‐bearing. A significant decrease in the number of C. albicans cells in the kidneys of NACOS‐6‐treated tumor‐bearing mice was observed 8 days after the fungal infection, or 15 days after the tumor transplantation. The candidacidal activity of polymorphonuclear leukocytes from NACOS‐6‐treated tumor‐bearing mice did not differ from that of NACOS‐6‐untreated tumor‐bearing mice. On the other hand, the candidacidal activities of both macrophages and T lymphocytes increased following administration of NACOS‐6 in the early phase of tumor‐bearing. The culture supernatant of T lymphocytes from NACOS‐6‐treated tumor‐bearing mice also potentiated the candidacidal activity of casein‐induced macrophages. An enhancement of natural killer cell activity of splenic lymphocytes obtained from NACOS‐6‐treated tumor‐bearing mice was also observed.
Increased Toxin Production by Vibrio cholerae Ol during Survival with a Green Alga, Rhizoclonium fontanum, in an Artificial Aquatic Environment
Authors: Islam Mohammad Sirajul
Published: November 2013

In the aquatic environment, the physiological state of Vibrio cholerae can be affected by various environmental conditions (e.g., sunlight, pH, temperature, competition with other bacteria for nutrients, etc.). The effect of these factors on the toxigenicity of V. cholerae was investigated. Toxin production by 5 toxigenic strains of V. cholerae incubated in laboratory microcosms containing Rhizoclonium fontanum was tested at different time intervals. The microcosms were exposed to sunlight, and the V. cholerae were in competition for nutrients with the resident bacterial flora of R. fontanum. The increase or decrease in toxin production by V. cholerae recovered at different time intervals was measured by ELISA and compared with the parent strains. Results of the study demonstrated an increase in toxin production by V. cholerae Ol during survival with R. fontanum. It is concluded that various environmental conditions in the aquatic environment affect toxin production by V. cholerae.
Electron Microscopic Observation of Crystals of Escherichia coli K‐12 Lipopolysaccharide
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Ito Hideo , Arakawa Yoshichika , Naito Setsuko
Published: November 2013

Previously we showed that Salmonella minnesota Re and Ra lipopolysaccharides (LPSs) and Escherichia coli K‐12 LPS formed three‐dimensional crystals, either hexagonal plates or solid columns, when they were precipitated by the addition of 2 volumes of 95% ethanol containing 375 mM MgCl2 and incubated in 70% ethanol containing 250 mM MgCl2, and stored at 4 C for 10 days. Later, Escherichia coli K‐12 LPS thus treated was found to form discoid crystals as well as hexagonal plate crystals and solid column crystals. Analysis by electron diffraction of the discoid crystals from the direction perpendicular to the basal plane showed that they consisted cf hexagonal lattices with the a axis of 4.62 Å. This result was quite the same as that of the hexagonal plate crystals. Electron micrographs of the edges of the discoid crystals revealed stacked sheets of the hexagonal plate crystals. From these results it was concluded that formation of the discoid crystals results from irregular overlapping of the hexagonal plate crystals.
Serotype Analysis of Group A Human Rotavirus Related to Acute Gastroenteritis in Winter in Gifu City
Authors: Kawamoto Hiroyoshi , Tanaka Hiroshi , Urasawa Shozo , Urasawa Tomoko , Taniguchi Koki
Published: November 2013

A total of 348 fecal specimens collected from children with acute gastroenteritis in Gifu city over three consecutive winter seasons (from November 1986 to March 1989) were examined for group A human rotavirus (HRV) by either a commercial test kit or a sandwich enzyme‐linked immunosorbent assay (sandwich ELISA). One hundred twelve of the 173 group A HRV‐positive specimens were further subjected to serotype determination by ELISA with four serotype‐specific monoclonal antibodies to VP7 (ELISA‐serotyping). Ninety‐one specimens (81.3%) were successfully serotyped: 41 (36.6%) were serotype 1, 13 (11.6%) serotype 2, 27 (24.1%) serotype 3, and 10 (8.9%) serotype 4. The serotypes of the remaining 21 (18.8°0) could not be determined. The predominant serotype of HRV that prevailed in Gifu city changed every winter: serotype 3 (63.4%) was most prevalent in the 1st winter, serotype 4 (42.9%) in the 2nd winter, and serotype 1 (64%) in the 3rd winter.
Fecal IgA Antibody Responses after Oral Poliovirus Vaccination in Infants and Elder Children
Authors: Nishio Osamu , Sumi Junko , Sakae Kenji , Ishihara Yuichi , Isomura Sin , Inouye Sakae
Published: November 2013

We investigated fecal IgA antibody responses after oral polyvalent poliovirus vaccination. Infants were given vaccines twice with an interval of 6 weeks. Specific IgA antibodies in the feces were determined by enzyme‐linked immunosorbent assay, and viruses were isolated in tissue cultures. We found that, after the first vaccination, antibody responses seemed to be elicited only against the serotypes of isolated viruses. After the second vaccination, however, antibodies were detected to all three serotypes with higher titers, suggesting that the first vaccination induced the immunologic memory. The IgA antibodies had virus‐neutralizing activity, and existed in the feces as both intact 11S and fragmented 4S molecules. Next, children were given the third vaccination 3 or 9 years later. Fecal IgA antibody responses were found to be poorer in elder children, while they responded with high serum neutralization titers. The secretory IgA memory seemed to last much shorter than the serum IgG memory.
Drug Resistance and R Plasmids in Pasteurella multocida Isolates from Swine
Authors: Yamamoto Junya , Sakano Tetsuya , Shimizu Mikio
Published: November 2013

A total of 163 strains of Pasteurella multocida isolated from swine were examined for drug resistance and R plasmids. Strains resistant to sulfadimethoxine (Sar), ampicillin (Apr), streptomycin (Smr), kanamycin (Kmr), and chloramphenicol (Cpr) were found in 93.9, 1.8, 16.6, 1.2, and 10.4%, respectively. There were two patterns of drug resistance (Sar and SarCpr) in isolates from nasal cavities, and five patterns (Sar, SarSmr, SarSmrCpr, SarSmrApr, and SarSmrKmrCpr) in isolates from pneumonic lung specimens. Two isolates studied were proved to carry a nonconjugative R plasmid pJY2 or pJY8 with other unidentified plasmids, respectively. pJY2 (3.6 megadaltons) encoding resistance to SarSmr had one cleavage site for EcoRI or HindIII endonuclease and two sites for PstI endonuclease. pJY8 (5.5 megadaltons) encoding resistance to Sar SmrKmrCpr had one EcoRI site and two PstI sites.
Early Development of CD8‐Negative and CD8‐Positive MHC‐Unrestricted Cytotoxic Cells Induced by Alloantigen Inoculation in Mice
Authors: Nishi Yasuaki , Hosokawa Tomohide , Aoike Akira , Kawai Keiichi , Kamahora Takeshi
Published: November 2013

Peritoneal cells (PC) in C57B1/6 (B6, H‐2b) mice receiving an intraperitoneal (i.p.) injection of allogeneic BALB/c (H‐2d) spleen cells demonstrated potent cytotoxic activity against syngeneic, xenogeneic, third‐party allogeneic tumors as well as H‐2d derived tumors. Maximum cytotoxic activity against various tumors other than H‐2d derived tumor, B16 (H‐2b) was elicited on day 3 post allosensitization and decreased drastically thereafter, whereas cytotoxic activity against P815 (H‐2d) peaked 3 days after the inoculation and maintained the peak activity thereafter. Surface phenotype of PC responsible for the cytotoxic activity against B16 was Thy‐1+/‐, Lyt‐2‐, L3T4−, asialo GM1 (AGM1)+, and that of PC against P815 was Thy‐1+, Lyt‐2+ (or Lyt‐2+/‐), L3T4~, AGM1+. These phenotypes showed similar phenotypes to the counterparts against B16 and against P815 in spleen cells induced by intravenous inoculation of alloantigen. When mice were pretreated i.p. with anti‐AGM1 antibody before the allosensitization, anti‐P815 cytotoxic activity in PC was completely diminished. Similar activity in spleen, however, was enhanced by i.v. treatment with the antibody before the i.v. inoculation of alloantigen. The data clearly demonstrate that in vivo inoculation of B6 mice with normal allogeneic cells induces “NK‐like” CD8‐ cytotoxic cells and “anomalous” CD8+ cytotoxic cells in PC.
Idiotypical Identity of IgG Myeloma Protein with Monoclonal IgM, Bence Jones Protein, and Fv Derived from One Patient
Authors: Miyagawa Nobuyuki , Morita Keiko , Azuma Takachika
Published: November 2013

Serum from a patient (KK) with IgG2‐λ myeloma was shown to contain multiple paraproteins corresponding to an IgM‐λ monoclonal protein (MMP), a λ‐type Bence Jones protein (BJP), and a 30 kDa component in addition to the IgG2 myeloma protein (GMP). These proteins possessed common idiotypic determinants, as judged by their monoclonal reactivity with rabbit anti‐GMP idiotype antibody (aId) in the immunofixation electrophoresis. Analysis with aId absorbed with either H or L chain of GMP revealed that the 30 kDa component shared both VH and VL with GMP and MMP. while BJP carried only the VL idiotype. The 30 kDa component, however, failed to react with antibody to either the μ, γ, α, K, or λ isotype, indicating that it had an Fv‐like molecular composition. These results suggest that myeloma cells of KK had diverged from the same precursor B cell clone to produce MPs of different isotypes and altered molecular constructions.
Attachment of Mycoplasma salivarium and Mycoplasma ovale to Glass Surfaces
Authors: Watanabe Tsuguo , Yukitake Hideharu
Published: November 2013

Mycoplasma salivarium ATCC 23064 and 24 other oral strains, and Mycoplasma orale ATCC 15539 and 22 other oral strains were examined quantitatively for attachment to glass surfaces by using ELISA method. Although all of the tested strains attached to glass surfaces, M. salivarium attached far more readily than M. orale. The results suggested that electrostatic bonds were involved in the attachment and that bivalent metal ions also played a role.
Dose‐Dependent Transplacental Infection of Murine Encephalomyelitis Virus GDVII in Gravid Uterus
Authors: Miyamae Takeo
Published: November 2013

Intrauterine infection of murine encephalomyelitis virus GDVII was investigated by the indirect immunofluorescence method. Sixty‐day‐old pregnant mice were inoculated with 5 gradated doses of the virus ranging from 0.001 to 1.5 MLD50 via i.v. route on the 10th day of gestation. The mice exsanguinated on postinfection day 8 showed high incidences of viral antigen in the uterine walls, placentas, and fetal subcutaneous tissues with doses of 0.1‐1.5 MLD50, regardless of the presence of maternal symptoms. Incidence of viral antigen‐positive cells in fetal brains was high with a dose of 0.1 MLD50, but not by the other doses. However, the brains of the stillborn and newly‐born mice derived from females infected by a dose of 0.5 MLD50 brought about still higher detection rates of viral antigen, as well as in the postpartum uteri. In effect, transplacental transmission of the virus was clearly demonstrated, and appeared to be dose‐dependent.
Elevated Expression of Proto‐Oncogenes during Interleukin‐5‐Induced Growth and Differentiation of Murine B Lineage Cells
Authors: Migita Masahiro , Yamaguchi Naoto , Katoh Shigeki , Mita Seiji , Matsumoto Ryoji , Sonoda Eiichiro , Tsuchiya Hiroyuki , Matsuda Ichiro , Tominaga Akira , Takatsu Kiyoshi
Published: November 2013

Intcrleukin 5 (IL‐5), a lymphokine produced by helper T cells, is involved in the regulation of growth and differentiation of B cells and other hematopoietic cells. To elucidate IL‐5‐mediated intracellular mechanisms, we have established IL‐5‐dependent and ‐independent murine early B cell lines, J6 and MJ88‐1, respectively, and examined the effect of IL‐5 on the expression of proto‐oncogenes during proliferation. Two‐ to 3.5‐fold increases in the levels of c‐myb, c‐myc, c‐fos, and c‐fms mRNA were observed in J6 cells, compared with those in MJ88‐1 cells. Further, a role of IL‐5 in the proto‐oncogene expression during differentiation was examined by using thymidine‐treated murine B‐cell chronic leukemia BCL1‐B20 cells with growth arrest. After 4‐day culture, the amount of IgM secreted from BCL1‐B20 cells was augmented 4‐6 fold in the presence of IL‐5. Although expression of c‐myb, c‐fos, and c‐fms mRNA did not change, only c‐myc mRNA expression was elevated within 30 min of stimulation with IL‐5 and reached a maximal level by 1 hr. Addition of phorbol 12‐myristate 13‐acetate (PMA) or IL‐4 to the culture of BCL1‐B20 cells inhibited both the IL‐5‐mediated augmentation of IgM secretion and the elevated expression of c‐myc mRNA. These findings suggest that the IL‐5 signal may be associated with the up‐regulation of c‐myc expression.
Molecular Cloning and Partial DNA Sequencing of the Collagenase Gene of Vibrio alginolyticus
Authors: Fukushima Jun , Takeuchi Hiroaki , Tanaka Eiichi , Hamajima Kenji , Sato Yayoi , Kawamoto Susumu , Morihara Kazuyuki , Keil Borivoj , Okuda Kenji
Published: November 2013

DNA fragments Vibrio alginolyticus chemovar iophagus, at least 7 kb in length, were ligated to Escherichia coli expression vectors. Three clones of Escherichia coli HB101 (pLCO‐1, pLCO‐2, pLCO‐3) were obtained by the colony immunoblotting method using anti‐collagenase antibody. In Escherichia coli, all these genes produced collagenase antigens which were detected with Western blotting. The amino acid sequence of chemically purified collagenase fragments was also analyzed. An approximately 2.5 kb DNA fragment of the pLCO‐1 clone was sequenced, and we found that portions of the deduced amino acid sequence were completely identical with the corresponding amino acid sequence of the chemically analyzed fragments. Therefore, it is highly probable that the gene studied in the present experiment is truly a collagenase structural gene.
Growth Physiology of Mycobacteria in Modified Dubos Liquid Medium
Authors: Tsukamura Michio
Published: November 2013

Although mycobacteria grow in Dubos liquid medium showing an arithmetic linear growth, the initial few days of growth were found to correspond to an ‘induction’ period. In this period, rapid increase of the amount of growth occurred, whereas increase of the number of colony‐forming units remained at a low level. This finding shows that the rapid increase of the amount of growth is accompanied by rapid death of multiplied bacteria. In a successive period, which was considered to correspond to the logarithmic growth phase, a 1:1 correspondence existed between the amount of growth and the number of colony‐forming units. The induction period is not considered to be a lag phase, in which the bacteria grow slowly, but a period of unbalanced relationship between the growth and the viability. Even when we inoculated different sizes of bacteria, the amounts of growth became similar in both inoculations after several days of incubation. However, the number of colony‐forming units remained always smaller in the use of small inocula than in the use of large inocula. In the use of small inocula, much more rapid increase of the amount of growth occurred. However, this rapid increase gave rise to rapid death of bacteria.
Permeability of Gentamicin and Polymyxin B into the Inside of Bacillus subtilis Spores
Authors: Fujita Yuko , Yasuda Yoko , Tochikubo Kunio
Published: November 2013

The penetration of gentamicin and polymyxin B into the inside of Bacillus subtilis spores was examined by an immunoelectron microscopy method with colloidal gold—immunoglobulin G (IgG) complex. The colloidal gold particles were located predominantly in the coat region of both gentamicin‐treated and polymyxin B‐treated spores and were hardly observed in the other regions, i.e., the cortex and core regions. When these antibiotic‐treated spores were subsequently treated with CaCl2, the number of gold particles bound to the coat region was greatly decreased. These results suggest that these two antibiotics are able to penetrate into the spore coat but not into the cortex or core, that is, the primary permeability barrier to them exists between the coat and the cortex regions.
The Occurrence of α(1 → 2) Linked N‐Acetylperosamine‐Homopolymer in Lipopolysaccharides of Non‐O1 Vibrio cholerae Possessing an Antigenic Factor in Common with Ol V. cholerae
Authors: Haishima Yuji , Kondo Seiichi , Hisatsune Kazuhito
Published: November 2013

Chemical analysis was carried out on lipopolysaccharides from Vibrio cholerae bio‐serogroup Hakata 487–85. The O‐specific chain of the phenol‐soluble lipopolysaccharides was demonstrated by 13C‐NMR spectroscopy and methylation analysis to contain a linear homopolymer of α(l→2) linked N‐acetylperosamine (4‐acctamido‐4,6‐dideoxy‐D‐mannopyranose), which was closely similar to but not identical to a linear α(l→2) linked N‐3‐deoxy‐L‐glycerotetronyl (S‐2,4‐dihydroxy‐butyryl) perosamine‐homopolymer constituting that of Ol Vibrio cholerae lipopolysaccharides.
Diversity of Matrix Protein in Subacute Sclerosing Panencephalitis and Measles Virus‐Infected Cells
Authors: Haga Takeshi , Yoshikawa Yasuhiro , Yamanouchi Kazuya
Published: November 2013

Expression of the viral matrix (M) proteins in Vero cells infected with 18 strains of subacute sclerosing panencephalitis (SSPE) virus and measles virus was examined by immunocytochemistry and Western blot analysis using an anti‐M monospecific serum and two sera against the M protein specific synthetic peptides. By immunocytochemistry using the anti‐M monospecific serum, M protein was detected in all of the virus‐infected cells regardless of cell‐free virus production. M proteins of the seven non‐productive strains were found to vary significantly in their epitope, in their reactivity to different assay systems, and in their molecular weight, whereas M proteins of the other 11 productive strains were detected consistently. These results suggest diversification of M protein of the non‐productive strains.
The Adhesion of Protein A‐Bearing Staphylococcus aureus Organisms to Soluble‐Staphylococcal‐Antigens‐Coated HeLa Cells Mediated by Specific Antibodies
Authors: Murai Miyo , Sakurada Junji , Seki Keiko , Ogasawara Masami , Arai Yoshiko , Edamura Keiko , Masuda Shogo
Published: November 2013

The adhesion of staphylococcal protein A (SpA)‐bearing Staphylococcus aureus Cowan I organisms to HeLa cells was enhanced by pretreatment of HeLa cells with staphylococcal extracellular antigens and antibodies to them. The adhesion of HLj, an SpA‐poor mutant derived from Cowan I, to HeLa cells was not enhanced by the same pretreatment of HeLa cells. Furthermore, the enhanced staphylococcal adhesion was inhibited by soluble SpA. The antigen(s) responsible for the enhanced staphylococcal adhesion was(were) heat stable. Pretreatment of HeLa cells with the mixture of staphylococcal extracellular antigens and antibodies to them also enhanced the adhesion of Cowan I. Similarly the adhesion of Cowan I was enhanced by pretreatment of HeLa cells with extracellular antigens of Pseudomonas aeruginosa and antibodies to them. These results indicated that cell‐bound SpA mediated the binding of S. aureus to immune complexes composed of extracellular bacterial products and antibodies to them bound to the surface of HeLa cells, and suggested another role of cell‐bound SpA as a co‐adhesin with other factors in infections due to S. aureus.
Comparative Characterization of Inducible and Virulent Vibrio parahaemolyticus Bacteriophages Having Unique Head Projections
Authors: Koga Tetsuro , Kawata Tomio
Published: November 2013

Phage TP1, induced from Vibrio parahaemolyticus K‐20 pilot strain by mitomycin C, exhibited a unique hexagonal head with knob‐like projections which covered the whole capsid and a noncontractile tail. The appearance of this phage was very similar to those of phages VP3 and VP6, isolated from seawater. The host range of phage TP1 was similar to those of phages VP3 and VP6. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) of the three phage particles revealed almost identical patterns with four major polypeptides with apparent approximate molecular masses: 78, 42, 37 and 34.5 kDa. On the basis of HindII digestion patterns on agarose gel electrophoresis, the lengths of phage TP1 and VP3 DNAs were estimated to be about 65 kilobase pairs (kbp) and that of VP6 DNA was about 74 kbp. The digestion patterns of all three phage DNAs by DraI, BamHI and MspI were very similar. The DNAs of TP1 and VP3 exhibited almost the same digestion patterns with HindIII and EcoRI, whereas the digestion patterns of VP6 DNA were significantly different from those of the former. From these findings, it seems likely that virulent phage VP3 is originated from a lysogenic phage, probably TP1, of V. parahaemolyticus.
Pattern of Shedding of Small, Round‐Structured Virus Particles in Stools of Patients of Outbreaks of Food‐Poisoning from Raw Oysters
Authors: Haruki Kosuke , Seto Yoshiyuki , Murakami Tsukasa , Kimura Teruo
Published: November 2013

The pattern of shedding of the small, round‐structured virus (SRSV) particles in the stools of patients who suffered from food‐poisoning due to raw oysters was investigated. The duration and concentration of fecal shedding of the SRSV particles were studied by electron microscopic examinations of stool specimens obtained during the course of illness to see a relation of viral shedding to day of illness. It was found that the fecal shedding of the SRSV particles occurred within five days of illness; thereafter, the concentration of the SRSV particles in feces rapidly decreased within a few days during the course of illness.
Isolation, and Morphological and Chemical Properties of an Autolysis‐Deficient Mutant of Clostridium botulinum Type A
Authors: Kinouchi Takemi , Takumi Kenji , Kawata Tomio
Published: November 2013

An autolysis‐deficient mutant was isolated from Clostridium botulinum type A 190L by treatment with ethyl methanesulfonate. The cell wall prepared from the mutant autolyzed at much slower rate than that from the parent strain, accompanying with much less liberation of both amino terminals and reducing groups. Electron microscopic observation revealed that the mutant strain was converted to short rod or curved spherical form with thickned cell walls when the growth temperature was shifted from 37 to 45 C. The mutant had a significantly larger amount of non‐peptidoglycan‐carbohydrate complexes than did the parent strain and became markedly resistant to the autolysin partially purified from the parent, compared with the parent strain. Furthermore, the mutant was fairly tolerant to killing by penicillin. These results suggest that the autolysis deficiency of the mutant was due not only to the deficient production of autolysin but also to the excess accumulation of carbohydrate in the cell wall.
Adherence to HEp‐2 Cells and Replication in Macrophages of Salmonella derby of Human Origin
Authors: Budiarti Sri , Hirai Yoshikazu , Minami Junzaburo , Katayama Sei‐ichi , Shimizu Tohru , Okabe Akinobu
Published: November 2013

Adherence to a HEp‐2 cell monolayer was tested for in four strains of Salmonella derby which were isolated from patients with diarrhea. One strain, SB1, was highly adherent and another strain, SB4, was nonadherent. The other two strains exhibited moderate adherence. Further in vitro study of invasion of HEp‐2 cells by S. derby and its replication in murine peritoneal macrophages was carried out using SB1 and SB4. Thin section electron micrographs revealed that SB1 invaded HEp‐2 cells but SB4 did not. The number of viable bacteria within macrophages was determined at intervals after inoculation of bacteria. The result indicates that SB1 can replicate in the macrophages but SB4 cannot. Flagella and fimbriae were compared by electron microscopy between SB1 and SB4, and their lipopolysaccharides and outer membrane proteins were also compared with each other by SDS‐polyacrylamide gel electrophoresis. The presence of a 41 kDa protein in the outer membranes of SB1 was only the difference detected, suggesting that this protein could be a factor required for adherence of this serovar to epithelial cells.
Plasmid Incidence Rate and Conjugative Chloramphenicol and Tetracycline Resistance Plasmids in Malaysian Isolates of Salmonella typhi
Authors: Phipps Maude , Pang Tikki , Koh Chong Lek , Puthucheary Savithri
Published: November 2013

Seven (6.1%) of 115 strains of Salmonella typhi isolated from Malaysian patients harbored a single large plasmid of 71 to 166 mD. Two of the seven plasmid‐bearing strains were resistant to chloramphenicol (Cm) and tetracycline (Tc) and they transferred Cm and Tc resistance traits to Escherichia coli K12 at frequencies from 1.6 × 10−7 to 1.9 × 10−6. Agarose gel electrophoresis provided evidence that the resistance traits were cotransferred on a conjugative plasmid. The significance and importance of these results are discussed.
The 106‐Kilobase Plasmid of Salmonella braenderup and the 100‐Kilobase Plasmid of Salmonella typhimurium Are Not Necessary for the Pathogenicity in Experimental Models
Authors: Horiuchi Sankichi , Goto Nobuichi , Inagaki Yoshio , Nakaya Rintaro
Published: November 2013

Among 1,041 clinical isolates (77 serovars) of Salmonella which had been derived from cases with acute enterocolitis, 601 (58%) contained one or more plasmids. Large serovar‐specific plasmids were seen in 95 of 307 isolates (31%) of Salmonella typhimurium, in 34 of 34 isolates (100%) of Salmonella enteritidis and in 36 of 38 isolates (94.7%) of Salmonella braenderup: the sizes of which were 100, 60 and 106 kilobases (kb), respectively. In order to determine the role of these plasmids in pathogenicity for enterocolitis, the plasmids were eliminated from some strains of S. braenderup and S. typhimurium and the pathogenicity of the plasmid‐less strains was compared with that of the parent strains by invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, lesion of mucosal tissue and the Sereny test. The virulence of all the plasmid‐less strains was as strong as that of the plasmid‐bearing strains in these pathogenicity assay systems. We therefore concluded that the 106‐kb plasmid of S. braenderup and the 100‐kb plasmid of S. typhimurium are not necessary for their pathogenicity in the experimental models: invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, and Sereny test.
Comparison of Protective Effects with Tetra‐Valent Glycolipid Antigens and Whole Cell‐Inactivated Vaccine in Experimental Infection of Leptospira
Authors: Masuzawa Toshiyuki , Suzuki Ryoma , Yanagihara Yasutake
Published: November 2013

The protective antigens (PAgs), glycolipid substance, were extracted from Leptospira interrogans serovars autumnalis, hebdomadis, australis and copenhageni, which were considered as main causal serovars of human leptospirosis in Japan, with chloroform‐methanol‐water (1:2:0.8, [vol/vol/vol]) solution. The tetra‐valent formalin‐inactivated leptospiral vaccine (Weil's disease and Akiyami combined vaccine) composed of the four serovars mentioned are used as vaccine to protect human from leptospiral infection in Japan. The protective effect, agglutinating antibody‐inducing activity and opsonin‐inducing activity of tetra‐valent PAgs were compared with those of vaccines now in use, which were supplied by two companies, Takeda Chemical Industries, Ltd., and Denka‐Seiken Co., in Japan. The tetra‐valent PAgs which contained 10 μg of each PAg protected hamsters and cyclophosphamide‐treated mice from lethal infection of serovar copenhageni and induced agglutinating antibodies against the four serovars in the same degrees as vaccines. These results suggested that the tetra‐valent PAgs might be useful as a component vaccine against leptospiral infection instead of formalized whole cells vaccines for human.
Role of L3T4+ and Lyt‐2+ T Cell Subsets in Protective Immune Responses of Mice against Infection with a Low or High Virulent Strain of Toxoplasma gondii
Authors: Nagasawa Hideyuki , Manabe Tetsuya , Maekawa Yoichi , Oka Mikio , Himeno Kunisuke
Published: November 2013

In order to elucidate the role of T cell subsets in protective immunity against infection with high virulent and low virulent strains of Toxoplasma gondii, monoclonal antibodies specific for T cell subsets were injected into mice before immunization or challenge infection. Treatment of mice with monoclonal antibody to either L3T4+ or Lyt‐2+ T cells before they were immunized with Toxoplasma cell homogenate prepared from high virulent RH strain tachyzoites markedly reduced survival after mice were challenged with low virulent bradyzoites of the Beverley strain. Thus, induction of protective immunity against bradyzoites of the Beverley strain requires the presence of both L3T4+ and Lyt‐2+ T cells. In contrast, mice injected with living bradyzoites of the low virulent Beverley strain after immunization with Toxoplasma cell homogenate acquired protective immunity against high virulent tachyzoites of the RH strain. Lyt‐2+ T cells alone appear to be final effector cells for protection against the challenge with high virulent RH strain tachyzoites, since treatment of the bradyzoite‐immune mice with anti‐Lyt‐2 antibody, but not anti‐L3T4 antibody, before challenge significantly increased mortality.
Detection of Influenza Viruses in Throat Swab by Using Polymerase Chain Reaction
Authors: Yamada Akira , Imanishi Jiro , Nakajima Etsuro , Nakajima Katsuhisa , Nakajima Setsuko
Published: November 2013

An assay protocal based on exploiting the polymerase chain reaction (PCR) for the direct detection of influenza virus in throat swab is described. By use of the mixture of HI and H3 primers, it was possible to determine the subtype of the influenza A viruses simultaneously. No visible band was detected after PCR of influenza B or A (H2N2) viruses with a pair of HI or H3 primers. The dilution experiment showed that the influenza viruses, as few as 1.3–6 plaque‐forming units, were sufficient for detecting the HA gene by PCR. All throat swab samples from which influenza viruses had been isolated by conventional method were also positively detected by PCR method.
Heat‐Killed Salmonella typhi Induces the Release of Prostaglandins and Leukotrienes from Mouse Macrophages
Authors: Pang Tikki , Devi Shamala , Puthucheary Savithri , Pawlowski Nicholas
Published: November 2013

Mouse macrophages pre‐labeled with [3H]arachidonic acid (20: 4) were shown to release metabolites generated by the lipoxygenase and cyclo‐oxygenase pathways following in vitro addition of heat‐killed Salmonella typhi. These metabolites were maximally released after 60–90 min of incubation and consisted of prostaglandins (85%), leukotriene C (6%), di‐HETEs, leukotrienes D and E (4%), mono‐HETEs (2%) and other metabolites (3%). Of the metabolites generated by the cyclo‐oxygenase pathway (prostaglandins), 6‐keto PGF1 α and PGE2 were generated at a ratio of 1.2 to 1. The significance and importance of these results are discussed.
Possible Presence of a Capsule in Branhamella catarrhalis
Authors: Ahmed Kamruddin , Rikitomi Naoto , Ichinose Akitoyo , Matsumoto Keizo
Published: November 2013

Clinical isolates of Branhamella catarrhalis from patients with respiratory infections were used in this study. Electron microscopic observation after treating Branhamella catarrhalis with immune serum and ruthenium red revealed the capsule. In the phagocytosis test, most organisms were not ingested by human polymorphonuclear neutrophils in the presence of normal rabbit serum (NRS), while organisms were primarily cell associated and apparently ingested in the presence of immunized rabbit serum (IRS). The capsule may be one of the virulence factors in this bacteria. This study demonstrates the possible presence of a capsule in Branhamella catarrhalis.
Colonization of Helicobacter pylori in the Gastric Mucosa of Mongolian Gerbils
Authors: Yokota Kenji , Kurebayashi Youichi , Takayama Yoshikazu , Hayashi Shunji , Isogai Hiroshi , Isogai Emiko , Imai Kouzou , Yabana Tsuyoshi , Yachi Akira , Oguma Keiji
Published: November 2013

Helicobacter pylori was orally inoculated into Mongolian gerbils. The organisms were able to colonize in the gastro‐mucosal layer of the gerbils, especially in those gerbils which had mucosal lesions caused by indomethacin treatment. The pathological changes developed by H. pylori infection were restricted to the stomachs, and only slightly inflammatory cells were observed.
Effect of Clostridium perfringens Alpha Toxin on Contraction of Isolated Guinea‐Pig Diaphragm
Authors: Sakurai Jun , Tsuchiya Yasunobu , Ochi Sadayuki , Fujii Yoshio
Published: November 2013

The effect of Clostridium perfringens alpha toxin on contraction induced by‐electric stimulation of isolated guinea‐pig diaphragm was investigated. The toxin inhibited electrically stimulated contraction of the tissue in a dose‐ and incubation time‐dependent manner. Tetrodotoxin resulted in no effect of the action of the toxin. Nifedipine dose‐dependently delayed the action of the toxin, but verapamil and diltiazem did not. On the other hand, treatment of the toxin with N‐acetylimidazole caused significant reduction of the inhibitory activity of the toxin on contraction, but did not cause significant loss of phospholipase C activity (PN activity) as measured by hydrolysis of p‐nitrophenylphosphorylcholine. The data showed that the toxin impairs contraction of isolated guinea‐pig diaphragm.
Human Immunodeficiency Virus Infection in Cells of Myeloid‐Monocytic Lineage
Authors: Ushijima Hiroshi , Dairaku Masatake , Honma Hitoshi , Yamaguchi Koushi , Shimizu Hiroyuki , Tsuchie Hideaki , Abe Kenji , Yamamoto Akiko , Hoshino Hiroo , Müller Werner E.G.
Published: November 2013

We established persistent infection with a strain of human immunodeficiency virus type I, HTLV‐IIIB, in a promyelomonocytic cell line, ML‐1 (CD4 antigen nearly negative and CD4 mRNA negative), and a promonocytic cell line, THP‐1 (CD4 antigen positive). Different reaction of giant cell formation was found after co‐cultivation of infected and uninfected cells of ML‐1, HL‐60, THP‐1 and U‐937 cell lines with uninfected and infected MOLT4 (a T‐lymphoma cell line).
Gene Transfer in Enteric Bacteria through the Formation of R‐Prime Plasmids by an RP4: : Mini‐Mu Element
Authors: Sarker Mahfuzur R. , Ahmed Zia U. , Rahman M.M.
Published: November 2013

Gene transfer in seven pathogenic enteric bacteria was studied using an RP4::mini‐Mu element, the plasmid pULB113. From the E. coli K‐12 host strain the plasmid could be efficiently transferred to these enteric bacteria, but its transfer back to E. coli K‐12 was not as efficient, being detected only in Shigella dysenteriae 1, S. flexneri and the ‘smooth’ variant of S. sonnei. In these three species, transposition of chromosomal fragments into the plasmid to produce R‐prime plasmid was also detected at a frequency of approximately 10−5. Transposition was random as suggested by the recovery at approximately the same frequency (10−5 to 10−6) of R‐primes involving 20 different auxotrophic markers from widely separated chromosomal locations. Formation of R‐prime plasmids expressing toxicity in the E. coli K‐12 recipient strain was also efficient in S. dysenteriae 1 but the toxin‐activity was rapidly lost from these R‐primes. In our experiments, the plasmid pULB113 incorporated relatively small amounts of chromosomal DNA as determined by restriction endonuclease digestion. For a Thy+ R‐prime that we analyzed, the amount of cloned DNA was approximately 15 kb.
T‐Cell Immune Response to Human Herpesvirus‐6 in Healthy Adults
Authors: Yakushijin Yoshihiro , Yasukawa Masaki , Kobayashi Yuzuru
Published: November 2013

The proliferative response of peripheral blood mononuclear cells (PBMC) from healthy adults to human herpesvirus‐6 (HHV‐6) was examined. It was found that PBMC from 13 of 14 HHV‐6‐seropositive adults apparently proliferated in response to stimulation with HHV‐6 antigen in contrast to the lack of response of cord blood mononuclear cells. In order to confirm the presence of HHV‐6‐specific memory T cells in the peripheral blood of healthy adults, we established HHV‐6‐specific T‐cell clones from an HHV‐6‐seropositive individual. CD4+ T‐cell clones generated from HHV‐6‐stimulated PBMC were found to proliferate upon stimulation with HHV‐6 in the presence of autologous antigen‐presenting cells, but not in response to herpes simplex virus type 1 antigen or mock‐infected control antigen, These results indicate that a T‐cell immune response against HHV‐6 infection is generally present in healthy adult populations.
Acute Phase Reactants in Leprosy
Authors: Vaishnavi Chetana , Agnihotri Nalini , Ganguly Nirmal Kumar , Kaur Surrinder , Kumar Bhushan
Published: November 2013

Serum levels of C‐reactive protein (CRP) and plasma levels of fibronectin (Fn) were studied in 74 untreated leprosy patients. CRP was detected by latex agglutination in 25.6% of the patients. A significant increase in Fn levels was seen in all the groups of leprosy patients, as compared to the controls.
Development of Monoclonal Antibodies Reacting against Mycobacterial 65 kDa Heat Shock Protein by Using Recombinant Truncated Products
Authors: Hara Isao , Sato Noriyuki , Matsuura Akihiro , Cho Joong‐moon , Weimin Qi , Torigoe Toshihiko , Shinnigk Thomas M. , Kamidono Sadao , Kikuchi Kokichi
Published: November 2013

A mycobacterial 65 kDa molecule is a member of the GroEL heat shock protein family. We developed mAbs reacting against recombinant 65 kDa protein by using a gene (pTB12) which encodes this protein. Three mAbs (B20, B97 and B167) reacted selectively with 65 kDa proteins of Mycobacterium tuberculosis, BCG and Mycobacterium leprae, although B20 and B167 may weakly react with a 15 kDa molecule of mammalian cells. One (B108) was obviously cross‐reactive between mycobacterial 65 kDa and the mammalian intracytoplasmic protein. We also developed deletion mutants of pTB12. The localization of these mAb‐defined epitopes was determined by using truncated proteins of the Mycobacterium tuberculosis 65 kDa molecule produced in E. coli. Immunohistochemical analysis showed that B20, B97 and B167 mAbs could detect this antigen in experimental granulomas induced by injection of BCG in the subcutaneous tissue of rats. These mAbs should be useful for analyzing the immunobiologic roles of mycobacterial 65 kDa molecules.
Toxin Profiles, and Cell‐Surface Properties of Salmonella Strains Isolated from Swedish Travelers
Authors: Baloda Suraj B. , Krovacek Karel
Published: November 2013

Toxin production, cell‐surface hydrophobicity and fibronectin‐binding properties of 21 Salmonella strains of different species, isolated from Swedish travelers to different parts of the world, were studied. Cell sonicate supernatants from blood agar grown cultures of 80% of the strains induced rabbit skin permeability reaction in the form of induration and/or blueing while 33% of the strains also produced cell necrotizing factor on rabbit skin. Four strains were negative in the rabbit skin permeability test, while only two were negative when tested on CHO cells. When cultured on blood agar, a majority of the strains (17/21) showed low cell‐surface hydrophobicity, showing no aggregation even at 1.5 M ammonium sulfate concentration in salt aggregation test (SAT), while only four strains showed high cell‐surface hydrophobicity. Furthermore, these strains could be classified as low fibronectin binders due to their poor interaction with fibronectin or its 29 kDa N‐terminal fragment.
Rapid Diagnosis of Cytomegalovirus Infections by Direct Immunoperoxidase Staining with Human Monoclonal Antibody against an Immediate‐Early Antigen
Authors: Eizuru Yoshito , Minematsu Toshio , Minamishima Yoichi , Ebihara Kazumasa , Takahashi Kota , Tamura Kazuo , Hosoda Kenji , Masuho Yasuhiko
Published: November 2013

Direct immunoperoxidase technique using a horseradish peroxidase (HRP)‐conjugated Fab' fragment of human monoclonal antibody (humab C7), designated HRP‐C7, was evaluated as a rapid diagnosis of cytomegalovirus (CMV) infection. A total of 138 clinical specimens consisting of 124 urine samples and 14 oral swabs were examined for CMV by the direct HRP‐C7 staining in comparison with conventional virus isolation. The number of CMV‐positive samples by each method was 40 (29.0%) for the former and 37 (26.8%) for the latter, respectively. By HRP‐C7 staining, CMV was identifiable within 24 hr after inoculation. By conventional isolation method, an average of 10.3 days had passed before cytopathic effect characteristic of CMV appeared in the cell culture. Some false‐positive and false‐negative cases were discussed in relation to toxicity of urine samples, storage of the samples, and amount of CMV in the sample. The sensitivity and specificity of HRP‐C7 method against conventional isolation method were 89.2% and 93.1%, respectively. Thus, HRP‐C7 staining is useful for a rapid diagnosis of CMV infections.
Quantitative Fluorometric Assay for Rapid Enzymatic Characterization of Bifidobacterium longum and Related Bifidobacteria
Authors: O'Brien Mark , Mitsuoka Tomotari
Published: November 2013

The quantitative, semi‐automated assay described here is an alternative characterization method allowing for highly sensitive and specific detection of bifidobacterial enzymes. Twenty strains of Bifidobacterium longum, including the type strain ATCC 15707, and type strains of 15 other Bifidobacterium species were enzymatically characterized using 20 4‐methylumbelliferyl conjugated substrates. Enzyme activities were determined by directly measuring the intensity of fluorescence derived from 4‐methylumbelliferone, a fluorescent metabolic by‐product. For this method, a Titertek® Fluoroskan II fluorometer was used. Enzymes included glycosidases, an esterase, phosphatase, sulphatase, and neuraminidase. B. longum showed strong activity (> 1,000 absolute fluorescence units, afu) for α‐L‐Arabino‐pyranosidase and α‐L‐Arabinofuranosidase, β‐D‐Fucosidase, α‐ and β‐D‐Galactosidase, α‐D‐Glucosidase, and α‐D‐Mannosidase. No activity (<50 afu) was observed for β‐D‐Cellobiosidase, α‐ and β‐L‐Fucosidase, β‐D‐Glucuronidase, β‐D‐Mannosidase, Neuraminidase and Sulphatase. Enzymatic activity profiles in other bifidobacteria were different according to the species. This assay is simple and rapid (6 hr). Special cultural requirements are unnecessary. Results are objective and quantitative. This assay may be a useful tool for bifidobacterial taxonomy.
Pharmacodynamic and Protective Properties of a Murine Lipopolysaccharide‐Specific Monoclonal Antibody in Experimental Pseudomonas aeruginosa Pneumonia in Mice
Authors: Oishi Kazunori , Sonoda Fuminari , Miwa Hideaki , Tanaka Hirofumi , Watanabe Kiwao , Matsumoto Keizo , Pollack Matthew
Published: November 2013

We employed a Pseudomonas aeruginosa mouse pneumonia model to evaluate the ability of a murine monoclonal antibody (MAb) specific for the O‐side chain of P. aeruginosa Fisher Immunotype‐1 lipopolysaccharide (LPS) to achieve and sustain therapeutic levels in plasma and lung tissue, reduce bacterial populations in the lung, and prevent pneumonia‐associated mortality. An IgG3 MAb (Y1–5A4) administered to mice i.v. over a dose range of 125–1,000 μg/mouse produced plasma and lung tissue levels at 2 hr of 61–507 μg/ml and 4.3–150 μg/g, respectively. The 1,000 μg MAb dose reduced bacterial counts in lung tissue (logio cfu/g±S.D.) and blood (log10cfu/ml+S.D.) 20 hr post‐treatment (18 hr post‐challenge) from 10.00+ 0.66 to 7.66±0.91 (P<0.01) and from 4.39±0.81 to <3.0, respectively. Administration of MAb to mice in doses of 125–500 μg 2 hr prior to a 3 × 50% lethal bacterial challenge produced significant protection against death, with a calculated 50% protective dose of 167 μg. Protection was noted following administration of 1,000 μg of MAb up to 6 hr after bacterial challenge (P<0.05, compared with untreated control). Histological examination of lung tissue from infected mice revealed less acute inflammation, necrosis, and hemorrhage in MAb‐treated compared with untreated control animals and greater localization of Pseudomonas antigen within the phagocytic cells in alveolar space. These findings document the in vivo therapeutic efficacy of an LPS‐specific IgG MAb in a murine model of acute P. aeruginosa pneumonia, based in part upon the achievability of effective MAb concentrations in plasma and lung tissue.
Experimental Study on the Relapse of Tuberculosis after the Termination of Antituberculous Chemotherapy Using Immunodeficient Nude Mice
Authors: Toyohara Mareichi
Published: November 2013

The effect of cellular immunity induced by tuberculous infection on bacteriological relapse after the termination of antituberculous chemotherapy was studied experimentally using immunodeficient nu/nu mice and immunocompetent dd mice. The efficacy of intensive chemotherapy was excellent even in nu/nu mice; tubercle bacilli in the organs decreased below the detectable limit, but formidable regrowth of bacilli was seen after the termination of chemotherapy. On the other hand, in the case of dd mice that established antituberculous cellular immunity through tuberculous infection, bacteriological relapse was generally very slight. It was concluded that bacteriological relapse was related closely with the established cellular immunity induced by the infected tubercle bacilli.
The Surface Hydrophobicity and Avirulent Character of an Encapsulated Strain of Klebsiella pneumoniae
Authors: Meno Vuko , Amako Kazunobu
Published: November 2013

In order to elucidate how virulence is controlled in encapsulated bacteria, some surface properties of an encapsulated but avirulent strain of Klebsiella pneumoniae, strain 277, were examined. Although strain 277 was heavily fimbriated, the fimbriae did not demonstrate an avirulent character and were not responsible for the surface hydrophobicity of this strain. The surface hydrophobicity was well correlated with the capacity of the bacteria to associate with polymorphonuclear cells. More bacteria with hydrophobic surfaces associated with the PMN than nonhydrophobic bacteria. The hydrophobic surface character of this strain was not affected by either trypsin treatment or extraction with salt solution. We assume that the capsule of strain 277 has more hydrophobic polysaccharides than that of the virulent strain. Some chemical modifications might therefore exist in the capsular polysaccharides of the avirulent strain.
A Sensitive Method for Detecting the Fermentation‐Inhibition Antibody to Mycoplasma pneumoniae
Authors: Okazaki Norio , Akema Riichiro , Takizawa Kinjiro
Published: November 2013

The fermentation‐inhibition (FI) test for Mycoplasma pneumoniae was improved by using a combination of guinea pig complement and gamma globulin‐depleted horse serum in place of unheated whole horse serum employed in the conventional assay system. As the test antigen for the new FI assay system, M, pneumoniae filtrated through a 3.0 μm membrane filter was used. Owing to the strong augmenting effect of guinea pig complement, the FI activity of rabbit immune serum was increased 32‐fold in the new system compared with the conventional system. Furthermore, IgM antibody, which is barely detectable by the conventional system, could easily be titrated by the new system. With this sensitive method, rapid rise of FI titer was clearly demonstrable in most children with acute M. pneumoniae infections, and a prevalence of FI or growth‐inhibitory antibody among healthy adults in Japan (82%) was revealed.