bluuebunny/crossref_metadata_2025 huggingface dataset:
An Activity Which Restores Theta Toxin Activity in Some Theta Toxin‐Deficient Mutants of Clostridium perfringens
Authors: Imagawa Tadashi , Higashi Yasushi
Published: November 2013

Group a mutants of Clostridium perfringens are deficient in theta toxin but release a dialyzable substance (“substance A”), which restores theta toxin activity to group b mutants, into a culture supernatant; group b mutants are defective in “substance A” release. “Substance A” activity appeared in the exponentially growing phase of group a mutants and disappeared in the stationary phase. “Substance A” activity was most stable at pH 5.0 and 0 C and even increased threefold in the first 5 hr, but gradually decreased during the following 15 hr. It was quickly inactivated at neutral and higher pHs at 0 C.
Conspicuous Ingestion of Staphylococcus aureus Organisms by Murine Fibroblasts In Vitro
Authors: Usui Akemi , Murai Miyo , Seki Keiko , Sakurada Junji , Masuda Shogo
Published: November 2013

A conspicuous adhesion of Staphylococcus aureus organisms to murine cutaneous fibroblasts and NIH/3T3 cells cultured in vitro and subsequent ingestion of S. aureus organisms by these fibroblasts are described. In the present experimental system, only fibroblasts‐adhering S. aureus organisms were efficiently ingested by fibroblasts unlike S. epidermidis and S. saprophytics. These findings might suggest a correlation between the pathogenesis of S. aureus and its intracellular localization in non‐professional phagocytes such as fibroblasts in a special reference to its higher pathogenicity than those of coagulase negative counterparts.
Adenovirus Infection and Specific Secretory IgA Responses in the Intestine of Infants
Authors: Nishio Osamu , Sakae Kenji , Ishihara Yuichi , Isomura Shin , Inouye Sakae
Published: November 2013

We investigated adenovirus (Ad) infection of the intestine and Ad group‐specific fecal IgA antibody responses in seven infants who were followed up from birth to 16 months to seven years of age. We isolated in tissue culture from fecal samples not only enteric Ad type 41 but also other Ads (types 2, 3, 5, 6, and 12). We also detected Ad antigens in the feces by ELISA at the times of infection with even non‐enteric Ads, suggesting that a large amount of antigens were produced in the intestine. We found that repeated Ad infections with different serotypes were occurring and there were good fecal IgA antibody responses at each time. The infection seemed usually mild or asymptomatic: only one out of 23 occasions of the detected infections required hospitalization.
Nucleotide Sequence of the Arginine Deiminase Gene of Mycoplasma hominis
Authors: Harasawa Ryô , Koshimizu Kaoru , Kitagawa Masanari , Asada Kiyozo , Kato Ikunoshin
Published: November 2013

The arginine deiminase gene of Mycoplasma hominis was amplified by the polymerase chain reaction, and its entire nucleotide sequence was determined. This gene consists of 1227 base pairs encoding 409 amino acids, and has 35.2% guanine plus cytosine content. Nucleotide sequence homologies of the arginine deiminase gene between M. hominis and M. arginini, and between M. hominis and M. orale were 82.1 and 80.8%, respectively, suggesting that this gene is highly conserved among arginine‐utilizing Mycoplasma species.
Impact of Bifidobacterium longum on Human Fecal Microflora
Authors: Benno Yoshimi , Mitsuoka Tomotari
Published: November 2013

The effects of the Bifidobacterium longum feedings for five weeks on the fecal microflora, water contents, pH values, ammonia concentration, and β‐glucuronidase activity were investigated in five healthy human volunteers. Although numbers of major bacterial groups of the fecal microflora were not changed by the bifidobacteria feedings, a remarkably decreasing number of lecithinase‐negative clostridia was observed. The percentage of lecithinase‐negative clostridia and bacteroides to the total bacterial numbers isolated were decreased during the feedings and numbers of C. paraputrificum and C. innocuum were reduced. A significant reduction of fecal pH values for the last week of the feeding was observed. Ammonia concentration and β‐glucuronidase activity in the feces during the feedings were significantly lower than those before or after the feedings. The oral supplement of B. longum may be introduced to improve the fecal properties such as fecal ammonia concentration and β‐glucuronidase activity, but not the composition of fecal flora.
Further Evaluation of the Pregnancy‐Linked Down‐Regulation of the Paternal Antigen‐Specific Splenic Cytotoxic T Lymphocyte Activity in Allogeneically Pregnant Mice
Authors: Parvin Mahbuba , Isobe Ken‐ichi , Goto Setsuko , Nakashima Izumi , Tomoda Yutaka
Published: November 2013

Cytotoxic T lymphocyte (CTL) activity directed against paternal alloantigen was examined in allogeneically pregnant mice using various allogeneic combinations. The spleen cells from pregnant C57BL/6 (H‐2b) mice mated with BALB/c (H‐2d) male mice generated less anti‐H‐2d CTL after in vitro sensitization than those from unpregnant or syngeneically mated C57BL/6 mice. Different allogeneic combinations including the incompatibility at only D region of H‐2 or minor histocompatibility loci were effective for downregulating the anti‐paternal CTL activity in pregnancy. The downregulation of anti‐paternal CTL activity induced by allogeneic pregnancy occurred at day 10 to day 18 of pregnancy, most extensively at day 14. The allogeneic pregnancy also downregulated the allogeneic CTL activities that had been amplified by injecting alloantigens before mating.
A Simple Method for Overproduction and Purification of p24 Gag Protein of Human Immunodeficiency Virus Type 1
Authors: Tanaka Naoto , Saitoh Atsushi , Nakata Atsuo , Shinagawa Hideo
Published: November 2013

A simple method for the overproduction in Escherichia coli and purification of major core protein p24 of human immunodeficiency virus type 1 (HIV‐1) was described. The gag‐pol region encoding p24, p15, and protease was fused to 3′ end of lacZ gene on plasmid. A LacZ‐Gag fusion protein, the major primary product, is designed to be cleaved by the HIV‐1 protease coexpressed through frameshifting. In fact, p24 and its immediate precursor, p25, were produced in the cells grown at 25 C, but not at 37 C. When the gag and pol frames were fused in‐frame to express the protease without frameshifting, the main product, a LacZ‐Gag‐Pol fusion protein, was efficiently processed to give p24 exclusively both at 37 C and 25 C, suggesting more efficient expression of the protease. Recombinant p24 was purified to near homogeneity by a simple three‐step procedure. The amino‐terminal sequence of the recombinant p24 was the same as that of p24 deduced from nucleotide sequence, indicating that correct processing occurred in E. coli by the coexpressed protease. The method described here provides a means to obtain a large amount of highly pure p24, which is useful for crystallographic and functional studies, preparation of specific antibody, and diagnostic and prognostic uses.
Prevalence of Antibodies to Pseudomonas pseudomallei Exotoxin and Whole Cell Antigens in Military Personnel in Sabah and Sarawak, Malaysia
Authors: Embi Noor , Suhaimi Adlan , Mohamed Rahmah , Ismail Ghazally
Published: November 2013

Sera from 420 military personnel serving in Sabah and Sarawk, Malaysia, were tested for antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens by enzyme‐linked immunosorbent assay procedure (ELISA). Data showed that 54.4% of serum samples were positive for antibodies to P. pseudomallei exotoxin and 65.7% were positive for antibodies to the whole cell antigens. Samples gave much lower titers for anti‐exotoxin antibodies compared to titers against crude whole cell antigens. The incidence of antibody to exotoxin was highest in the age groups ranging from 26 to 32 years, where the positive rates were higher than 40% and 30% for military personnel serving in Sarawak and Sabah, respectively.
The Protective Activity of Tea Catechins against Experimental Infection by Vibrio cholerae O1
Authors: Toda Masako , Okubo Sachie , Ikigai Hajime , Suzuki Tatsuo , Suzuki Yumiko , Hara Yukihiko , Shimamura Tadakatsu
Published: November 2013

Tea catechins inhibited the fluid accumulation induced by cholera toxin in sealed adult mice. The catechins also reduced fluid accumulation by Vibrio cholerae O1 in ligated intestinal loops of rabbits. These findings suggest that tea catechins may possess protective activity against V. cholerae O1.
Comparison of Secretory Acid Proteinases fromCandida tropicalis, C. parapsilosisandC. albicans
Authors: Sono Eiko , Masuda Tomoko , Takesako Kazutoh , Kato Ikunoshin , Uchida Katsuhisa , Murayama Somay Yamagata , Yamaguchi Hideyo
Published: November 2013

Acid proteinases secreted byCandida tropicalisandC. parapsilosiswere newly isolated. Their physico‐chemical and enzymatic properties of molecular weight, pH stability, isoelectric points, specific activity, and N‐terminal amino acid sequences were determined and compared with those of aC. albicansacid proteinase. The two acid proteinases secreted byC. parapsilosiswere found to be new enzymes in their molecular weights. The acid proteinases fromC. tropicalisandC. parapsilosisshowed lower activity at neutral pH, less resistance to neutral and alkaline pH than that fromC. albicans, and a half or a third of the specific activity of theC. albicansenzyme. These differences seemed to be associated with the difference of pathogenesis betweenCandidaspecies. Of the 31 N‐terminal amino acids, the enzymes of these threeCandidaspecies revealed 12 homologous amino acids.
Demonstration of Low Molecular Weight Polypeptides Associated with Small, Round‐Structured Viruses by Western Immunoblot Analysis
Authors: Oishi Isao , Yamazaki Kenji , Kimoto Tatsuo , Minekawa Yoshiichi
Published: November 2013

Small, round‐structured viruses (SRSV) were detected in 14 of 300 fecal specimens obtained from patients with acute gastroenteritis by electron microscopy. These SRSV strains were morphologically indistinguishable from one another. While 11 of these strains had a single usual major structural protein with molecular weight of 63,000 (63K) daltons (p63), interestingly, three strains possessed a single major structural protein with molecular weight of 33K daltons (p33). Treatments of p63‐SRSV with proteolytic enzymes or denaturating reagents did not affect the molecular weight of p63, and the p33 was not detectable by Western immunoblot in the ultracentrifugal supernatant of the p63‐SRSV suspension. These results suggest that the p33 is neither a definitive subunit of p63 nor disintegrated component derived from the p63‐SRSV but a novel polypeptide of SRSV. Immune electron microscopy and Western immunoblot analyses indicated that p63‐ and p33‐SRSVs may share an antigenic determinant(s).
Species Dependency of In Vitro Macrophage Activation by Bacterial Peptidoglycans
Authors: Nagao Shigeki , Akagawa Kiyoko S. , Okada Fumito , Harada Yuji , Yagawa Katsuro , Kato Keijiro , Tanigawa Yoshinori
Published: November 2013

The effect of various bacterial cell wall components on in vitro biological function of murine peritoneal exudate macrophages was evaluated. We examined four different parameters of metabolic activity and monokine secretion. Peritoneal exudate macrophages from rats and guinea pigs, all of the strains tested, were stimulated by whole bacterial cell wall preparations, purified bacterial cell wall peptidoglucans, its water‐soluble peptidoglycan fragments, muramyl dipeptides and amphipathic substances. Murine peritoneal exudate macrophages were activated by amphipathic substances of gram‐positive bacteria. However, macrophages from mice, irrespective of strains, were not stimulated in the in vitro assay systems by purified bacterial cell wall peptidoglycan, water‐soluble bacterial peptidoglycan fragments or muramyl dipeptides. These results suggest that macrophage activation by bacterial peptidoglycan in vitro is animal species specific.
The Patterns and Transmissibility of Antibiotic Resistance among Clinical Strains of Pseudomonas aeruginosa
Authors: Palillo Eleanor S. , Salleh Mohd Azib
Published: November 2013

Four hundred and ninety‐eight predominantly pyocin‐type 10 clinical strains of Pseudomonas aeruginosa were analyzed for resistance to carbenicillin, cefoperazone, cefotaxime, ceftazidime, gentamicin, amikacin and netilmicin. Based on NCCLS‐recommended MIC breakpoints, 245 strains were found to be resistant, of which 41.6% were resistant to carbenicillin, 38% to gentamicin, 37.8% to netilmicin, 26.3% to cefoperazone, 17.9% to cefotaxime, 0.6% to amikacin and none to ceftazidime. Quadruple resistance to carbenicillin, cefoperazone, gentamicin and netilmicin was the most frequent pattern observed. Resistance to older antibiotics (kanamycin, streptomycin and tetracycline) and to mercuric chloride were also common. Conjugation experiments suggested that self‐transmissible and non‐transmissible plasmids occurred in at least 66 strains.
Augmentation of Bovine Leukemia Virus (BLV)‐Specific Lymphocyte Proliferation Responses in Ruminants by Inoculation with BLVenv‐Recombinant Vaccinia Virus: Their Role in the Suppression of BLV Replication
Authors: Ohishi Kazue , Suzuki Hidemi , Yasutomi Yasuhiro , Onuma Misao , Okada Kosuke , Numakunai Shigeru , Ohshima Kan‐ichi , Ikawa Yoji , Sugimoto Masanobu
Published: November 2013

Lymphocyte proliferation responses were investigated in sheep and cattle, in which the replication of bovine leukemia virus (BLV) had been known to be suppressed by inoculation with recombinant vaccinia virus (rVV) expressing BLV envelope glycoprotein (gp60). Enhanced lymphocyte proliferation responses were observed in animals inoculated with rVV, regardless of whether they were naive or BLV carriers. These responses were roughly inversely correlated to the growth of BLV in the peripheral blood leukocytes. In contrast, there was no apparent correlation between humoral immune response and BLV growth. Based on these results, it was suggested that rVV rendered its suppressive effect of BLV replication primarily via augmentation of cell‐mediated immunity.
Changes in the Susceptibility of 12‐O‐Tetradecanoylphorbol 13‐Acetate (TPA)‐Treated HL‐60 Cells to Staphylococcal Leukocidin
Authors: Morinaga Naoko , Kato Iwao , Noda Masatoshi
Published: November 2013

Susceptibility of human promyelocytic leukemia HL‐60 cells to staphylococcal leukocidin following treatment of cells with 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) was examined. TPA treatment for 6 hr rendered the cells very resistant transiently to leukocidin. There was no change in binding of leukocidin to the cells, but leukocidin‐induced 45CaCl2 influx, phospholipase A2 and C activities were inhibited. Further incubation with TPA rendered the cells sensitive again and then more sensitive than original HL‐60 cells following increase of the binding, and leukocidin‐induced activities described above appeared again. Those cells treated with TPA for more than 18 hr started to differentiate to macrophages morphologically and functionally. These data suggest that the differentiated cells were more sensitive than original HL‐60 cells because of increased binding of leukocidin and that treatment of TPA for 6 hr may transiently impair the signal transduction system of leukocidin after binding of leukocidin to the specific receptor of the cell membrane. Using these TPA‐treated cells, it was shown in this report that calcium influx, phospholipase A2 and C activities were important to induce cytotoxic action of leukocidin after binding of leukocidin to specific receptors on the cells.
Detection of Various Variant Verotoxin Genes in Escherichia coli by Polymerase Chain Reaction
Authors: Lin Zaw , Kurazono Hisao , Yamasaki Shinji , Takeda Yoshifumi
Published: November 2013

We constructed common primers for the polymerase chain reaction to detect the genes for various Verotoxins reported, that is, VT1 (or SLT‐I), VT2 (or SLT‐II), VT2vha, VT2vhb, SLT‐IIv (or VT2vp1, VTe) and SLT‐IIva (or VT2vp2). A total of 80 Verocytotoxin‐producing Escherichia coli strains isolated from humans, domestic animals and meats gave a positive result by PCR with the designed common primers. Digestion by restriction endonucleases BglII and EcoT14I of the amplicon of the VT2vp2 gene gave specific bands of the expected sizes, but not of the amplicons of other VT genes, suggesting a possible method for identification of the VT2vp2 gene. Application of the PCR with the designed primers in diagnostic and epidemiological studies on VTEC infection is also discussed.
Induction of Acute Arthritis in Mice by Peptidoglycan Derived from Gram‐Positive Bacteria and Its Possible Role in Cytokine Production
Authors: Onta Tokio , Sashida Masami , Fujii Noriyuki , Sugawara Shunji , Rikiishi Hidemi , Kumagai Katsuo
Published: November 2013

The activities of a water‐soluble peptidoglycan fragment derived from Staphylococcus epidermidis (SEPS) were examined as to their role in proliferation of spleen mononuclear cells (SMNC) from various strains of mice, the production of cytokines in vitro, and the induction of an inflammatory reaction in vivo. The proliferation of SMNC from C3H/HeN, C57BL/6, AKR, DBA/2, and ddY mice in reaction to SEPS in vitro showed a peak on day 3 and was greater than that of SMNC from BALB/c mice. The cells of SMNC from C3H/HeN mice responsive to SEPS were indicated to be mainly macrophages. A time kinetics experiment showed a coincidence in the proliferation of SMNC in reaction to SEPS and the detection of colony‐stimulating factor (CSF) activity. Interleukin 2 (IL‐2) activity was not detected during the incubation periods. When SEPS was administered to mice, much stronger mRNA transcripts of granulocyte‐macrophage (GM)‐CSF were detected in the lungs of C3H/HeN mice than in BALB/c mice. On the other hand, the amounts of IL‐1 and PGE2 produced by SMNC of BALB/c mice stimulated by SEPS were greater than those produced in C3H/HeN mice. SEPS was confirmed to induce arthritis in BALB/c mice, but not in C3H/HeN mice. Our findings suggest that the production of GM‐CSF is involved in the in vitro proliferation of SMNC in reaction to SEPS and that along with IL‐1 and PGE2 production, contributes to the inflammation by SEPS in vivo.
The Gene Encoding the Heat‐Stable Enterotoxin of Vibrio cholerae Is Flanked by 123‐Base Pair Direct Repeats
Authors: Ogawa Akira , Takeda Tae
Published: November 2013

The heat‐stable enterotoxin (O1‐ST) gene (sto) was cloned from chromosome of the strain GP156 of Vibrio cholerae O1 (Inaba, El Tor) in Escherichia coli K‐12, and its nucleotide seqence was determined. The nucleotide sequence of sto was very similar to that of NAG‐ST gene (stn) of V. cholerae non‐O1. Both sto and stn were flanked by 123‐base pair direct repeats which had at least 93% homology to one another and included some inverted repeats. All the strains of V. cholerae, V. mimicus, V. metschnikovii, V. hollisae and Yersinia enterocolitica examined by colony hybridization had the direct repeat sequence regardless of ST‐gene possession.
A Comparative Study on T Cell Receptor Vβ Gene Usages: Spleen Cells from the Non‐Obese Diabetic (NOD) Mouse and Its Non‐Diabetic Sister Strain, the ILI Mouse, and Infiltrating T Cells into Pancreata of NOD Mice
Authors: Koide Yukio , Kaidoh Toshiö , Yanagawa Tatsuo , Yoshida Takato O.
Published: November 2013

We analyzed the usage of T cell receptor (TCR) Vβ genes of spleen cells of NOD mice in comparison with those of its non‐diabetic sister strain ILI mice which show no insulitis and (ILI × NOD)F1 mice. The quantitative polymerase chain reaction (PCR) method revealed that PCR Vβ repertoires of these mice are indistinguishable. This is consistent with our previous observation that ILI mice share the same H‐2 class II genes with NOD mice. PCR method also revealed that the Vβ transcript of infiltrating T cells into pancreas of NOD mice was not restricted but was rather diverse. The role of TCR repertoire in the development of insulitis was discussed.
Release of Prostaglandin E2 by Human Mononuclear Cells Exposed to Heat‐Killed Salmonella typhi
Authors: Yusof Wan Noraini Wan , Nagaratnam Menaka , Koh Chong‐lek , Puthucheary Savithri , Pang Tikki
Published: November 2013

Human mononuclear cells pre‐labeled with [3H]arachidonic acid were shown to release metabolites following in vitro addition of heat‐killed Salmonella typhi (HKST). The amount of label released was significantly higher than that seen with live S. typhi (LST). Addition of increasing amounts of HKST resulted in an increased release of metabolites. Enzyme immunoassay of the culture supernatants revealed that the bulk of the metabolite released was prostaglandin E2 (PGE2). Leukotriene B4 (LTB4) and leukotriene C4 (LTC4) were not detectable in the culture supernatants. The significance and implications of these results are discussed.
The Structure of the Chloramphenicol Resistance Gene on a Transferable R Plasmid from the Fish Pathogen, Pasteurella piscicida
Authors: Kim Eun‐heui , Aoki Takashi
Published: November 2013

The chloramphenicol resistance gene (pp‐cat) was cloned from a transferable R plasmid of Pasteurella piscicida, pSP9351, and the sequence of the gene was determined. Subcloning and deletion analysis localized the resistance gene, pp‐cat, to within a 2.3 kb HincII‐BamHI fragment. The fragment as a probe hybridized with the type I chloramphenicol acetyltransferase (CAT) gene and did not hybridize to CAT types II, III, and CAT‐VA. The fragment hybridized to transferable R plasmids encoded with resistance to chloramphenicol, which were detected from P. piscicida isolated in different years. Nucleotide sequences of the coding and flanking regions of pp‐cat (2031 bp) identified an open reading frame coding type I CAT of a molecular mass of about 25,000 Da. Comparison analysis of the sequences outside the cat open reading frame showed also that pp‐cat has homology, in part, with the gene that coding for the endonuclease EcoRII and those that flank the cat gene derived from the Acinetobacter baumannii chromosome.
Japanese Patients with Chronic Fatigue Syndrome Are Negative for Known Retrovirus Infections
Authors: Honda Mitsuo , Kitamura Katsuhiko , Nakasone Tadashi , Fukushima Yoshiko , Matsuda Shunji , Nishioka Kusuya , Matsuda Juzo , Hashimoto Nobuya , Yamazaki Shudo
Published: November 2013

Although chronic fatigue syndrome (CFS) is known to be the syndrome that begins with an acute flu‐like illness that may be due to the exposure to an infectious agent, there has been no convincing evidence on the causative agents. Recently, human T‐lymphotropic virus type II (HTLV‐II)‐like virus has been reported to be associated with the CFS by using HTLV Western blot analysis and polymerase chain reaction. However, some investigators could not detect HTLV‐II by indirect immunofluorescence analysis. Lately, CFS patients have been reported in Japan. We detected all 30 tested patients with CFS were seronegative for HTLV‐II, HTLV‐I and HIV by specific peptide ELISA and Western blot. Further, PCR analysis was negative for HTLV‐II and retrovirus was not detected by coculture method with patients' PBMC. Thus, known human retrovirus infections do not cause a CFS in Japan.
Characterization of Vibrio cholerae O139 Synonym Bengal Isolated from Patients with Cholera‐Like Disease in Bangladesh
Authors: Higa Naomi , Honma Yasuko , Albert M. John , Iwanaga Masaaki
Published: November 2013

Vibrio cholerae O139 (synonym Bengal), a novel serovar of V. cholerae, is the causative agent of large outbreaks of cholera‐like illness currently sweeping India and Bangladesh. Eight randomly selected V. cholerae O139 isolates were studied for their biological properties, which were compared with those of V. cholerae O1 and other V. cholerae non‐O1. The V. cholerae O139 isolates were characterized by the production of large amount of cholera toxin, hemagglutination, weak hemolytic properties, resistance to polymyxin B, lysogeny with, and production of, kappa type phage (4/8 isolates only), and resistance to both classical and El Tor‐specific phages. Thus, V. cholerae O139 isolates had an overall similarity with V. cholerae O1 El Tor.
Effect of Heparin on Hemagglutinin of Herpes Simplex Virus Type 1
Authors: Noda Masahiro , Inaba Yuji , Seno Masato , Kanamoto Yasuo , Tokumoto Shizuyo
Published: November 2013

Heparin inhibited the hemagglutinin activity of herpes simplex virus (HSV) type 1. The minimal inhibitory concentration of heparin required to inhibit 8 hemagglutination (HA) U of HSV ranged from 0.005 to 0.01 U/ml. Mouse erythrocytes failed to combine with the HA inhibitory factor of heparin. On the other hand, mouse erythrocytes treated with heparinase had greatly reduced agglutinability by HSV. Virus‐heparin complex formation was observed by sedimenting heparin with the virus particles.
Dual Infection of HIV‐1 and HTLV‐I in South India: A Study on a Patient with AIDS‐Related Complex
Authors: Koyanagi Yoshio , Yoshida Tsutomu , Suzuki Michihiko , Uma Alagappan , Ananthasubramaniam Lalitha , Ramajayam Subbiah , Yamamoto Naoki
Published: November 2013

A dually HIV‐and HTLV‐infected ARC patient was found by serological studies in South India. These viruses were isolated and molecular study showed that the patient had both HIV‐1 and HTLV‐I but not HIV‐2 and HTLV‐II. In addition to this, 9 other dually infected persons which include another full‐blown AIDS case have been identified as on July 1993 in South India. Our findings provide an opportunity to clarify geographical distribution of these human retroviruses.
Cholera Toxin (Choleragen) ‐ Polymorphonuclear Leukocyte Interactions: Effect on Migration in Vitro and FcγR ‐ Dependent Phagocytic and Bactericidal Activity
Authors: Niemialtowski Marek , Klucinski Wlodzimierz , Malicki Konrad , De Faundez Irma Spohr
Published: November 2013

PMNL leukocytosis is a feature common to many types of infectious and inflammatory diseases. How PMNL are recruited to tissues is not yet clear although it is a question that has considerable clinical importance. We investigated the function of PMNL which migrated through an artificial barrier (Chinese hamster ovary (CHO) cells, collagen and nylon cloth membrane) subjected to CT or choleragenoid treatment toward plain medium (the same RPMI in the upper and lower chamber) or medium containing chemotactic factor (fMLP or LPS or ZAS). CT treatment significantly (P<0.01) reduced the FcγR expression on the surface of PMNL. The PMNL functions, namely, migration, phagocytic activity and intracellular killing of staphylococci, also have been reduced significantly (P<0.01). FcγR expression and some functions of PMNL that migrate to chemoattractants were reduced, irrespective of the presence or absence of CT; however, the inhibitory effect of CT on PMNL function was observed only when PMNL migrate to the lower chamber without chemotactic factor. On the other hand choleragenoid treatment of CHO cells did not have any significant influence on PMNL function and FcγR expression. In conclusion, our experiments demonstrate that CT reduces EAFc rosetting and the FcγR‐dependent phagocytic and bactericidal activity of bovine blood PMNL.
Effects of Cytochalasins B and D on Staphylococcus aureus Adherence to and Ingestion by Mouse Renal Cells from Primary Culture
Authors: Murai Miyo , Seki Keiko , Sakurada Junji , Usui Akemi , Masuda Shogo
Published: November 2013

Cytochalasin B (CB) and cytochalasin D (CD), inhibitors of microfilament function of host cell, were examined for their effects on Staphylococcus aureus Cowan I adherence to and ingestion by several types of the hyperosmolarity‐tolerant (HOT) cells obtained from primary culture of mouse kidney. Staphylococcal adherence to the HOT cells with epithelial appearance was extraordinarily enhanced by the treatment of those cells with both 5 μg/ml of CB and CD. In particular, staphylococci adhered to the periphery rather than the center of each cytochalasintreated cell. Staphylococcal ingestion by all types of the HOT cells was markedly inhibited by CD in spite of the enhanced adherence. Contrary to our expectation, inhibition by CB was incomplete, and the enhanced adherence of staphylococci to CB‐treated cells resulted in the enhanced ingestion.
Genetic Transformation in Helicobacter pylori
Authors: Tsuda Masataka , Karita Mikio , Nakazawa Teruo
Published: November 2013

Genetic transformation in Helicobacter pylori was investigated by using its chromosomal and plasmid DNAs. Six out of the eight strains exhibited the natural competence for incorporation of H. pylori chromosomal DNA, and all the strains incorporated the donor DNA efficiently by washing and concentrating the cells, with a glycerol solution. The much higher frequency of transformation was obtained in each strain by means of electroporation. Electroporation experiments were also conducted by use of the recombinant DNAs consisting of the H. pylori and Escherichia coli plasmids as the donors, and the occurrence of the homologous recombination was demonstrated between the incoming H. pylori plasmid‐derived region and the corresponding region of the originally residing plasmid in H. pylori.
Drug Resistance and Broad Geographical Distribution of Identical R Plasmids of Pasteurella piscicida Isolated from Cultured Yellowtail in Japan
Authors: Kim Eun‐Heui , Aoki Takashi
Published: November 2013

An MIC test of 12 chemotherapeutic agents performed on 175 strains of Pasteurella piscicida collected from cultured yellowtail (Seriola quinqueradiata) in different areas of Japan from 1989 to 1991 revealed 152 strains (87%) with resistance to combinations of ampicillin (AP), chloramphenicol (CP), kanamycin (KM), nalidixic acid (NA), sulfamonomethoxine (SA), tetracycline (TC), and/or trimethoprim (TMP). The remaining 23 strains were sensitive to all the drugs tested: AP, cefazolin, CP, florfenicol (FF), furazolidone, KM, NA, novobiocin, SA, streptomycin, TC, and TMP. FF showed the most effective antibacterial activity against P. piscicida with MICs ranging from 0.004 to 0.6 μg/ml. One hundred and forty‐nine of the 152 resistant strains carried transferable R plasmids encoding one of the Cp Km Sa Tc, Km Sa Tc, Km Sa, and Sa resistance. The most common resistance marker of transferable R plasmids identified in P. piscicida was Km Sa Tc. R plasmids encoding three different resistant markers were very similar on the basis of their digestion patterns with restriction endonucleases. There was homology among the DNAs of nine transferable R plasmids selected. Our findings suggest that multiple drug resistant strains of P. piscicida carrying transferable R plasmids with the same DNA structure are common in yellowtail farms and that the R plasmid has been retained within the P. piscicida population without change in their DNA structure according to geography and year.
Release of the Outer Membrane Vesicles from Vibrio cholerae and Vibrio parahaemolyticus
Authors: Kondo Katsuhiko , Takade Akemi , Amako Kazunobu
Published: November 2013

We found numerous small vesicles released from the cell by thin sectioning of the plate culture of Vibrio cholerae and V. parahaemolyticus fixed with the freeze‐substitution technique. From the broth media of exponentially growing bacteria we could collect the vesicles by the centrifugation but not enough without fixation. The vesicles are encompassed with a membrane structure similar to the outer membrane of these bacteria. The anti‐O (Inaba) serum reacted with the surface of the vesicles and the inside of the vesicle are generally filled with an electron‐dense mass.
Age‐Associated Increase of CD5+ B Cells in the Liver of Autoimmune (NZB × NZW) F1 Mice
Authors: Ohteki Toshiaki , Abo Toru , Kusumi Akinori , Sasaki Takeshi , Shibata Shinobu , Seki Shuhji , Kumagai Katsuo
Published: November 2013

The liver has been demonstrated to be a major site for extrathymic differentiation of T cells. In this study, an identification of CD5+ B cells, which are responsible for the onset of autoimmune disease by virtue of autoantibody production, was performed in autoimmune (NZB × NZW) F1 mice. An age‐associated increase of CD5+ B cells was demonstrated in the liver of these mice. Although CD5+ B cells (i.e., CD5+IgM+ and CD5+B220+) constituted a minor population of hepatic mononuclear cells (MNC) (<5%) when mice were young (8 weeks), a large population of CD5+ B cells (10 to 30% of whole MNC) was identified in the liver of mice aged 25 to 30 weeks after the onset of disease. Such age‐dependent increase of CD5+ B cells was not observed in any other strains including NZB, NZW, C3H/He and BALB/c mice. The phenotype of hepatic CD5+ B cells was the same as that of CD5+ B cells in the peritoneal cavity and spleen, showing dull‐CD5, bright‐IgM and dull‐B220. High levels of CD5+ B cells were observed in the peritoneal cavity and liver, but not in the spleen nor in any other lymphoid organs in mice aged 30 weeks. Radioimmunoassay of autoantibodies in the 5‐day culture supernatants demonstrated that hepatic MNC were unable to produce any amounts of IgM‐ and IgG‐autoantibodies against double‐stranded DNA and single‐stranded DNA, despite the increased proportion of CD5+ B cells. On the other hand, peritoneal exudate cells produced only IgM‐, but not IgG‐, autoantibodies, whereas splenic cells were able to produce both IgM‐ and IgG‐autoantibodies. These results suggest that the liver might support the generation of the most primitive CD5+ B cells in these mice and that such generation increases as a function of age, probably resulting in the onset of autoimmune disease.
Development and Evaluation of Capture Enzyme‐Linked Immunosorbent Assays for Detection of Immunoglobulin G and M Antibodies to Group A Streptococcal Antigens
Authors: Hayashi Shunji , Yokota Kenji , Takizawa Yoshihiko , Tomizawa Isao , Nejime Tetsuya , Oguma Keiji
Published: November 2013

Capture enzyme‐linked immunosorbent assays (ELISAs) were developed to detect immunoglobulin G and M antibodies to group A streptococcal (GAS) antigens, streptolysin O, streptokinase, and group A carbohydrate. The sensitivities and the specificities of the IgM capture ELISAs to each GAS antigen were high enough to distinguish the patients with GAS infections (diagnosed as GAS pharyngitis or scarlet fever) from the control groups (healthy people and patients with pharyngitis from whom GAS could not be isolated). On the other hand, the specificities of the IgG capture ELISAs were not very effective in diagnosis of GAS infections. When the capture ELISA and an indirect ELISA detecting IgM antibodies to group A carbohydrate were compared, false‐positive reactions due to rheumatoid factor occurred in the indirect ELISA, but did not occur in the capture ELISA. These results indicate that the capture ELISA works better than the indirect ELISA in detecting the IgM antibody, and that the IgM capture ELISA to GAS antigen provides a rapid and highly reliable serodiagnosis for GAS infections employing only a single serum.
Isolation of Scrapie Agent from the Placenta of Sheep with Natural Scrapie in Japan
Authors: Onodera Takashi , Ikeda Toshio , Muramatsu Yasukazu , Shinagawa Morikazu
Published: November 2013

A five‐month‐pregnant Suffolk sheep histologically diagnosed as spontaneous scrapie was studied. Western blot analysis was performed with rabbit serum against the sheep scrapie‐associated fibrils (SAF). In the proteinase K (pk)‐treated parental brain and spleen samples, three major bands (15 K, 18 K, and 23 K) were detected. These major bands were not detected from the placenta. Infectious agents were isolated in mice from the brain samples but not from the placental homogenates. In another case of a three‐month‐pregnant Corriedale sheep without any clinical sign of, but histologically diagnosed as scrapie, was also studied in a similar approach. In the parental brain samples, three major bands (15 K, 18 K and 23 K) were detected. SAF protein was not detected in the parental spleen and placenta. No bands reactive with the antiserum were detected in any other samples from the fetal brain and spleen in both cases. However, infectious agents were isolated in mice from both brain and placental homogenates. Since the placenta is an important site of natural infection, it is worthwhile to study these tissues for the epidemiological study of scrapie infection.
Soluble CD23 in Human Immunodeficiency Virus Type 1 Infected Patients
Authors: Hober Didier , Ajana Faiza , Boniface Michel , Estrada Rocio , Lobert Pierre Emmanuel , Sartiaux Claudine , Mouton Yves , Wattré Pierre , Maniez‐Montreuil MichèLe
Published: November 2013

The level of sCD23 produced in the course of human immunodeficiency virus (HIV) infection was measured in patients grouped according to the Centers for Disease Control by using an immunoradiometric assay. Soluble CD23 was evaluated in supernatants of peripheral blood mononuclear cell (PBMC) (106 cells/ml) stimulated by phytohemagglutinin (PHA). Compared with healthy controls (m±S.D. = 1.0 ±0.34 U/ml, n = 7), higher values were observed in some of the patients of group II (asymptomatic) (m±S.D. = 2±1.33, n = 9) and some of the patients of group IV (AIDS) (m±S.D. = 1.3 ±1.40, n = 8). Those results prompted us to compare the plasma levels of sCD23 in group II and group IV HIV‐infected patients and in healthy individuals. Soluble CD23 plasma levels in healthy patients (n = 42) ranged from 0 to 1.5 U/ml (m±S.D. = 0.9±0.33), in group II patients (n = 17) from 0 to 3 U/ml (m±S.D. = 0.92±0.83) and in group IV patients (n =73) from 0 to 2.9 U/ml (m±S.D. = 1.15±0.71). The differences between the patients and the healthy individuals were not statistically significant but individual sCD23 values higher than 2 U/ml were obtained in 6% of the group II patients and 16.7% of the group IV patients. Increased values of sCD23 were obtained in plasma from patients with secondary infectious diseases (groups IV‐C1 and IV‐C2) and from patients without secondary infectious diseases (group II, group IV‐A and group IV‐B). Elevated values of sCD23 were detected even in patients with low counts of CD4+ T cells and CD8+ T cells in their peripheral blood. sCD23 has numerous activities including control of IgE synthesis and cytokine‐like properties. Our results show a disarray of sCD23 in HIV‐infected patients which could be involved in drug reactions, allergic manifestations and the IgE‐level increase. Further investigations should attempt to define the role of sCD23 in clinical manifestations of HIV infection.
Transovarial Transmission of Chikungunya Virus by Aedes albopictus Mosquitoes Ingesting Microfilariae of Dirofilaria immitis under Laboratory Conditions
Authors: Zytoon Eiman M. , El‐Belbasi Hussein I. , Matsumura Takeo
Published: November 2013

Female Aedes albopictus mosquitoes of the Miki strain were experimentally fed on defibrinated sheep blood containing 5× 107 PFU of chikungunya virus and 20,000 microfilariae of Dirofilaria immitis per milliliter. Fully engorged mosquitoes transmitted the virus to a small percentage of the F1 progeny, but females of the F1 generation did not transmit the virus to the F2 progeny. The control mosquitoes that ingested the virus without microfilariae did not transmit the virus to their eggs, larvae, or pupae in the F1 or F2 generations. These results showed that A. albopictus of this strain that concurrently ingested the virus and microfilariae transmitted the virus by the transovarial route under experimental conditions.
A Positive Correlation between the Precursor Frequency of Cytotoxic Lymphocytes to Autologous Epstein‐Barr Virus‐Transformed B Cells and Antibody Titer Level against Epstein‐Barr Virus‐Associated Nuclear Antigen in Healthy Seropositive Individuals
Authors: Kusunoki Yoichiro , Huang Hua , Fukuda Yasuko , Ozaki Kyoko , Saito Mayumi , Hirai Yuko , Akiyama Mitoshi
Published: November 2013

A limiting dilution analysis was established to determine the precursor frequency (PF) of cytotoxic lymphocytes against autologous B cells transformed with the Epstein‐Barr virus (EBV). This method was found to detect mainly self‐restricted T‐cell activity and little non‐self‐restricted cytotoxicity. The mean PF in 21 healthy EBV‐seropositive persons was 1.4 × 10‐3 (range: 0.03 × 10‐3 to 8.7 × 10‐3) for peripheral blood mononuclear cells, whereas 4 samples of mononuclear cells obtained from umbilical cord blood had PFs below 0.007 × 10‐3. A positive correlation was observed between the PF and serum antibody titers against EBV‐associated nuclear antigen among the seropositive persons.
Mechanism of Inhibition of DNA Synthesis by 1‐β‐D‐Arabinofuranosyl‐E‐5‐(2‐Bromovinyl)uracil
Authors: Suzutani Tatsuo , Machida Haruhiko , Honess Robert W.
Published: November 2013

The mechanism of the inhibitory action of 1‐β‐D‐arabinofuranosyl‐E‐5‐(2‐bromovinyl) uracil triphosphate (BV‐araUTP) on DNA synthesis by Escherichia coli DNA polymerase I Klenow fragment was studied. Acting as a chain terminator, BV‐araUTP inhibited DNA synthesis by Klenow fragment more effectively than 2′, 3′‐dideoxythymidine triphosphate (ddTTP). However, the incorporation sites of BV‐araU monophosphate were restricted at consecutive dTMP sequence whereas ddTMP was incorporated at every dTMP site.
Anti‐Human IgE Monoclonal Antibodies Recognizing Epitopes Related to the Binding Sites of High and Low Affinity IgE Receptors
Authors: Takemoto Hiroshi , Nishimura Shinji , Kosada Yumi , Hata Satoshi , Takagi Shin , Hosoi Susumu , Ezumi Kiyoshi , Ide Misao , Harada Shigenori
Published: November 2013

Anti‐human IgE monoclonal antibodies (mAbs) were produced and eight clones recognizing epitopes on native IgE were selected. Epitopes were mapped by a competitive inhibition enzyme‐linked immunosorbent assay, Western blotting and a multi‐pin peptide technology. Four sites (one each in the Cε1, Cε2, Cε2/Cε3 junction and Cε3) were recognized by the mAbs. The relationship between the four epitopes and the binding sites of high and low affinity IgE receptors (FcεRI and FcεRII, respectively) was studied using a monovalent Fab fragment of each mAb as a binding inhibitor. The IgE‐FcεRII binding was clearly inhibited by the mAb recognizing the Cε2/Cε3 junction, suggesting that FcεRII binds to a rather limited area around the Cε2/Cε3 junction. The IgE‐FcεRI binding, on the other hand, was scarcely inhibited by any single mAb. However, the binding was inhibited when the epitope in Cε2 was blocked simultaneously with that at the Cε2/Cε3 junction or with that in Cε3, indicating that these three distinct epitopes are related to the FcεRI binding sites. When these three epitopes were shown in the stereograph of human IgE, the FcεRI binding area was spread largely on the groove side between Cε2 and Cε3 domains. These results suggest that FcεRI acquires the high affinity through multiple bindings.
Inflammatory and Phagocytic Response to Experimental Campylobacter jejuni Infection in Mice
Authors: Pancorbo Pedro L. , Gallego Aurelia M. , de Pablo Manuel , Alvarez Carmen , Ortega Elena , de Cienfuegos Gerardo Alvarez
Published: November 2013

After intraperitoneal inoculation with Campylobacter jejuni BALB/c, Swiss and DBA mice show a peritoneal inflammatory response of different intensity. Only BALB/c mice have a strong peritoneal response. Simultaneous intraperitoneal inoculation of C. jejuni plus FeCl3 increase both inflammatory response and phagocytic activity in Swiss mice, without production of diarrhea. Some thermostable compounds of C. jejuni have a very strong chemotactic activity against peritoneal cells of mice, whereas a diffusible, thermolabile and glutaraldehyde‐resistant factor has an inhibitory effect over murine peritoneal cell phagocytosis. Bactericidal activity of peritoneal cells increased after in vitro re‐challenge with C. jejuni. Bacteremia is present in all the mice strains tested, but the clearance is quick in DBA and slow in BALB/c and Swiss mice. These experiments confirm that in mice, peritoneal non‐specific mechanisms of defense, such as macrophages, play an important role in order to control C. jejuni infection.
Identification of Hepatitis C Virus Subtype 3b (HCV‐3b) among Japanese Patients with Liver Diseases Using Highly Efficient Primers for Reverse Transcription‐Polymerase Chain Reaction
Authors: Doi Hisaya , Yoon Seitetsu , Homma Morio , Hotta Hak
Published: November 2013

With a reverse transcription‐polymerase chain reaction method that utilized highly efficient primers, the NS5 region of hepatitis C virus (HCV) genome could be amplified from all of the 24 serum samples randomly obtained from Japanese patients with liver diseases. Subtype analysis on the basis of nucleotide sequence similarities of the amplified fragments revealed that 2 out of 24 Japanese patients (8.3%) were infected with the recently identified subtype HCV‐3b, which had been thought to be rare in Japan.
Cross‐Reactions of Human Lupus Autoantibodies with 8‐Methoxypsoralen Photomodified DNA Fragments
Authors: Arjumand Sadia , Ali Asif
Published: November 2013

DNA fragments of around 200 base pair (average size) have been covalently crosslinked with 8‐methoxypsoralen under 365 nm UV light. The photoadduct, induced antibodies in rabbits with a titer of > 1:12,800 by direct bindng ELISA. Binding data showed that the induced antibodies are conformation‐specific recognizing restricted conformational change at site of crosslinking. Human autoantibodies against DNA, bound not only to native DNA but to the photomodified DNA fragment as well. In addition, binding patterns of SLE sera obtained from different patients were remarkably similar, indicating the recognition of altered conformation of the modified polymer by naturally occurring SLE anti‐DNA autoantibodies.
Adhesion of Staphylococcus aureus to Bovine Mammary Epithelial Cells Induced by Growth in Milk Whey
Authors: Mamo Wubshet , Fröman Gunnar
Published: November 2013

Staphylococcus aureus strains isolated from bovine intramammary infection (mastitis) were tested for adhesion to bovine mammary epithelial cells after growth in milk whey or TSB. Bacteria grown in milk whey adhered more efficiently to mammary gland epithelial cells in vitro than the corresponding homologous bacteria grown in TSB. Trypsin treatment of milk whey‐grown S. aureus had no effect on their adherence. Whereas, pretreatment with periodate significantly decreased bacterial adherence capacity. Periodate treatment of TSB‐grown bacteria had no effect on adhesion to the mammary gland epithelial cells.
Mycobacterium malmoense Isolated from Soil
Authors: Saito Hajime , Tomioka Haruaki , Sato Katsumasa , Tasaka Hiromichi , Dekio Satoshi
Published: November 2013

Mycobacterium malmoense was isolated from a soil sample, and biological, biochemical, antigenic, and genetic characteristics of the isolate were described. This is the first report of isolation of this organism in Japan.
The Distribution of G (VP7) and P (VP4) Serotypes among Human Rotaviruses Recovered from Japanese Children with Diarrhea
Authors: Kaga Eiji , Iizuka Masahiro , Nakagomi Toyoko , Nakagomi Osamu
Published: November 2013

Both the G (VP7) and P (VP4) serotypes of human rotaviruses collected over a 10‐year period from Japanese children with diarrhea were determined by recently‐developed polymerase chain reaction‐based typing assays. The combination of G1 and P8 was found in 65.2% and the combination of G2 and P4 was found in 15.2%. For the rest of the specimens, only a few other combinations occurred and their relative frequencies were less than 10%. The viruses carrying P9 were always associated with G3 as is the prototype strain AU‐1.