bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular Characterization of an atl Null Mutant of Staphylococcus aureus
Authors: Takahashi Junko , Komatsuzawa Hitoshi , Yamada Sakuo , Nishida Tetsuya , Labischinski Harald , Fujiwara Tamaki , Ohara Masaru , Yamagishi Jun‐ichi , Sugai Motoyuki 
Published: November 2013

Abstractatl is a gene encoding a bifunctional peptidoglycan hydrolase of Staphylococcus aureus. The gene product of atl is a 138 kDa protein that has an amidase domain and a glucosaminidase domain, and undergoes processing to generate two major peptidoglycan hydrolases, a 51 kDa glucosaminidase and a 62 kDa amidase in culture supernatant. An atl null mutant was isolated by allelic replacement and characterized. The mutant grew in clusters and sedimented when grown in broth culture. Analysis of peptidoglycan prepared from the wild type and the mutant revealed that there were no differences in muropeptide composition or in glycan chain length distribution. On the other hand, the atl mutation resulted in pleiotropic effects on cell surface nature. The mutant cells showed complete inhibition of metabolic turnover of cell wall peptidoglycan and revealed a rough outer cell wall surface. The mutation also decreased the amount of protein non‐covalently bound to the cell surface and altered the protein profile, but did not affect proteins covalently associated with the cell wall. Lysis of growing cells treated with otherwise lytic concentration of penicillin G was completely inhibited in the mutant, but that of non‐growing cells was not affected by the mutation. The atl mutation did not significantly affect the ability of S. aureus to provoke an acute infection when inoculated intraperitoneally in a mouse sepsis model. These results further support the supposition that atl gene products are involved in cell separation, cell wall turnover and penicillin‐induced lysis of the cells.
Serological Evidence of Transmission of Human Influenza A and B Viruses to Caspian Seals (Phoca caspica)
Authors: Ohishi Kazue , Ninomiya Ai , Kida Hiroshi , Park Chun‐ho , Maruyama Tadashi , Arai Takaomi , Katsumata Etsuko , Tobayama Teruo , Boltunov Andrei N. , Khuraskin Lev S. , Miyazaki Nobuyuki 
Published: November 2013

AbstractSeroepidemiological surveillance of influenza in Caspian seals (Phoca caspica) was conducted. Antibodies to influenza A virus were detected in 54% (7/13), 57% (4/7), 40% (6/15) and 26% (11/42) of the serum samples collected in 1993, 1997, 1998 and 2000 by enzyme‐linked immunosorbent assay (ELISA). In an hemagglutination‐inhibition (HI) test using H1‐H15 reference influenza A viruses as antigens, more than half of the examined ELISA‐positive sera reacted with an H3N2 prototype strain A/Aichi/2/68. These sera were then examined by HI test with a series of naturally occurring antigenic variants of human H3N2 virus, and H3 viruses of swine, duck, and equine origin. The sera reacted strongly with the A/Bangkok/1/79 (H3N2) strain, which was prevalent in humans in 1979–1981. The present results indicate that human A/Bangkok/1/79‐like virus was transmitted to Caspian seals probably in the early 1980s, and was circulated in the population. Antibodies to influenza B virus were detected by ELISA in 14% (1/7) and 10% (4/42) serum samples collected from Caspian seals in 1997 and 2000, respectively. Our findings indicate that seal might be a reservoir of both influenza A and B viruses originated from humans.
Extracellular ATP Regulates Cell Death of Lymphocytes and Monocytes Induced by Membrane‐Bound Lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium
Authors: Into Takeshi , Okada Kazutaka , Inoue Nobuo , Yasuda Motoaki , Shibata Ken‐ichiro 
Published: November 2013

AbstractThe cytotoxicities of lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium to a lymphocytic cell line, MOLT‐4, and a monocytic cell line, HL‐60, was upregulated by ATP added extracellularly in a dose‐dependent manner. These lipoproteins induced ATP release and plasma membrane permeability increase in these cell lines. In addition, periodate‐oxidized ATP, an antagonist for P2X purinergic receptors, suppressed the cytotoxicity of the lipoproteins, suggesting the possibility that P2X receptors for ATP play crucial roles in the cytotoxicity. Activation of caspase‐3 induced by the lipoproteins, which was assessed by the cleavage of the synthetic substrate DEVD‐pNA and the endogenous substrate poly(ADP‐ribose) polymerase, was also upregulated and downregulated by extracellular ATP and periodate‐oxidized ATP, respectively. On the basis of these results, this study suggests that mycoplasmal lipoproteins induce the permeability increase in lymphocytes and monocytes, by which ATP is released, and the ATP regulates the cytotoxicities of the lipoproteins to the cells, possibly by interaction with ATP receptors such as P2X purinergic receptors.
Monitoring of Active HHV‐6 Infection in Bone Marrow Transplant Recipients by Real Time PCR; Comparison to Detection of Viral DNA in Plasma by Qualitative PCR
Authors: Ihira Masaru , Yoshikawa Tetsushi , Suzuki Kyoko , Ohashi Masahiro , Suga Sadao , Horibe Keizo , Tanaka Naoko , Kimura Hiroshi , Kojima Seiji , Kato Koji , Matsuyama Takaharu , Nishiyama Yukihiro , Asano Yoshizo 
Published: November 2013

AbstractTwelve (46%) of the 26 patients had human herpesvirus 6 (HHV‐6) viremia after bone marrow transplant (BMT). All isolates were recovered from the samples obtained at 2 weeks after BMT. The sensitivity and the specificity of detection of viral DNA in plasma by qualitative polymerase chain reaction (PCR) for monitoring active virus replication were 92% and 97% respectively. Moreover, the positive (85%) and negative (99%) predictive values were also high. The patients with HHV‐6 viremia showed a clear peak in HHV‐6 DNA in peripheral blood mononuclear cells (PBMCs) at 2 weeks after BMT, which was measured by real time PCR. The virus DNA level in PBMCs between the two groups (patients with viremia and patients without viremia) was statistically different at 2 weeks after BMT (P = 0.033). In patients with HHV‐6 viremia, mean HHV‐6 DNA copy number was higher in the samples collected at 2 weeks after BMT than the samples collected at any other time period.
Phosphorylation of Cytokeratin 17 by Herpes Simplex Virus Type 2 US3 Protein Kinase
Authors: Murata Takayuki , Goshima Fumi , Nishizawa Yuji , Daikoku Tohru , Takakuwa Hiroki , Ohtsuka Kenzo , Yoshikawa Tetsushi , Nishiyama Yukihiro 
Published: November 2013

AbstractWe previously reported the establishment of an HEp2 cell line which expresses the US3 protein kinase (PK) of herpes simplex virus type 2 (HSV‐2) upon induction with IPTG. Here we report that expression, phosphorylation and ubiquitination of cytokeratin 17 (CK17) are enhanced in US3‐expressing HEp2 cells. In vitro kinase and co‐immunoprecipitation assays provided evidence that US3 PK directly phosphorylates CK17. Expression of US3 PK caused a significant decrease in filamentous staining of CK17, suggesting that phosphorylation of CK17 by US3 PK causes a disruption of intermediate filaments. Our observations suggest a role for US3 in the regulation of CKs and intermediate filaments in cells. Moreover, we found that infection of a keratinocyte‐derived cell line, A431, with a US3‐deficient virus, results in cytopathic effects that are morphologically distinct from those induced by wild‐type and revertant viruses, suggesting that US3 PK may be important for interaction between HSV‐2 and peripheral epithelial cells.
Interferon Regulatory Factor 1 in Mycobacterial Infection
Authors: Yamada Hiroyuki , Mizuno Satoru , Sugawara Isamu 
Published: November 2013

AbstractIn order to understand the role of IRF‐1 in the development of murine tuberculosis in vivo, IRF‐1 knockout mice were infected with Mycobacterium tuberculosis by placing them in the exposure chamber of an airborne infection apparatus. These knockout mice developed multifocal necrotic lesions in the lung, liver and spleen tissues and died of disseminated tuberculosis within 43 days of infection. Compared with the levels in wild‐type mice, the pulmonary inducible NO synthase (iNOS) mRNA expression level was significantly lower, but IL‐18 and IL‐6 mRNA levels were higher. There was no statistically significant difference in the expression of IFN‐γ and TNF‐α mRNA between the IRF‐1 knockout and wild‐type mice. IRF‐1 is indirectly responsible for iNOS mRNA expression and plays an important role in the pathogenesis of murine tuberculosis.
Extracellular Proteinase and Phospholipase Activity of Three Genotypic Strains of a Human Pathogenic Yeast, Candida albicans
Authors: Sugita Takashi , Kurosaka Shoko , Yajitate Maki , Sato Hitomi , Nishikawa Akemi 
Published: November 2013

AbstractStrains of a human pathogenic yeast, Candida albicans, have (A) intronless, (B) intron‐containing, and (C) a mixture of intron‐containing and intronless 26S rRNA genes. To elucidate the significance of these three genotypes in pathogenesis, we measured two major virulence factors, extracellular proteinase and phospholipase activity, in 56 clinical isolates of C. albicans, and investigated the relationship between genotype and enzymatic activity. The genotype B strains had significantly higher proteinase and phospholipase activities than genotypes A or C. These results suggest that to understand the pathogenesis of C. albicans, the genotypes should be considered.
Development of a Mucosal Complex Vaccine against Oral Salmonella Infection in Mice
Authors: Harada Hiroko , Nishikawa Fumiko , Higashi Nobutaka , Kita Eiji 
Published: November 2013

AbstractWe examined the immunogenicity of a Salmonella enterica complex vaccine (CV), consisting of flagellin and polysome purified from serotype Typhimurium LT2. CV plus cholera toxin (CT), in three oral doses given at 7‐day intervals, conferred complete protection on C57BL/6 mice against lethal oral infection with a wild‐type strain. It elicited mucosal IgA>IgG2a>IgG1 and systemic IgG2a>IgG1>IgA antibodies to flagellin and polysome, and delayed footpad response (DFR) to both antigens. In Peyer's patches (PPs) and lamina propria (LP), IgA was produced under a Th1‐dominant environment; CD4T cells from produced interleukin (IL)‐2, interferon (IFN)‐γ, and IL‐10 by stimulation with salmonella extract. On the same protocol, flagellin plus CT induced flagellin‐specific mucosal and systemic IgA and IgG1 antibodies, CD4T cells producing IL‐10 and IFN‐γ in PPs and LP, and only minimal levels of flagellin‐specific DFR. Polysome plus CT induced polysome‐specific mucosal and systemic IgG2a in addition to IgG1 and IgA antibodies, CD4+T cells producing IFN‐γ and IL‐2 in PPs and LP, and polysome‐specific DFR. These two vaccines, however, conferred at most 50–60% survival rates. Our results suggest that polysomes in CV provide effective adjuvant activity for the induction of both mucosal and systemic Th1‐biased responses toward flagellin.
Selection of Human Immunodeficiency Virus Type 1 Variants with an Insertion Mutation in the p6gag and p6pol Genes under Highly Active Antiretroviral Therapy
Authors: Ibe Shiro , Shibata Naomi , Utsumi Makoto , Kaneda Tsuguhiro 
Published: November 2013

AbstractWe detected several types of human immunodeficiency virus type 1 (HIV‐1) variants with an insertion mutation in the p6gag and p6pol genes in eight of twenty‐two (36.4%) patients who possessed drug‐resistant viruses under highly active antiretroviral therapy (HAART). It was characteristic that a conserved proline‐rich motif “PTAPP” in the N‐terminus of p6gag protein was completely or partially duplicated in all cases. Five among the eight cases were retrospectively investigated in terms of the occurrence of dynamic change in the gag gene between the inserted and wild‐type HIV‐1 in the course of HAART. The longitudinal analysis revealed the following: 1) The inserted‐type viruses were selected over the wild‐type during HAART in three cases in which the both types coexisted in the beginning of the therapy. 2) In two cases in which the inserted‐type HIV‐1 alone was detected before the beginning of HAART, the inserted‐type HIV‐1 alone was continuously detected during the therapy. The inserted‐type HIV‐1 was also detected in four of thirty‐nine (10.3%) therapy‐naive patients. However, the frequency of inserted‐type HIV‐1 detection in the HAART‐receiving patients is significantly higher than that in the therapy‐naive patients (P=0.02). These results suggest that this type of insertion mutation is a polymorphism of the p6gag and p6pol genes, however, it consequently gave an advantage on proliferation and/or survival of the HIV‐1 variant under the presence of antiretroviral drugs.
Achievement of Complete Mycological Cure by Topical Antifungal Agent NND‐502 in Guinea Pig Model of Tinea Pedis
Authors: Uchida Katsuhisa , Tanaka Takashi , Yamaguchi Hideyo 
Published: November 2013

AbstractWe examined the therapeutic effect of a 1% cream preparation of NND‐502, a novel topical antifungal agent, in a guinea pig tinea pedis model produced by infecting the plantar skin of guinea pigs with Trichophyton mentagrophytes. Animals developing tinea pedis were divided into two groups: an untreated control group and a treated group. In the latter group, after confirming infection had been established, the infected animals were topically treated with the NND‐502 cream once daily for one week. The animals were reared in a clean environment free from exposure to exogenous dermatophytes. At one week (5 weeks post‐infection), 6 weeks (10 weeks post‐infection) and 16 weeks (20 weeks post‐infection) after completion of the treatment, plantar skin samples were taken from a certain number of both groups of animals. The results demonstrated that all of the animals in the untreated control group and none of those in the treated group were culture‐positive in this animal model of tinea pedis. The topical treatment with NND‐502 achieved a mycological cure. Thus NND‐502 can be considered a promising candidate as a new anti‐dermatophytic agent for topical use.
Seroprevalence ofBartonella henselae, Toxoplasma gondii, FIV and FeLV Infections in Domestic Cats in Japan
Authors: Maruyama Soichi , Kabeya Hidenori , Nakao Ruriko , Tanaka Shigeo , Sakai Takeo , Xuan Xuenan , Katsube Yasuji , Mikami Takeshi 
Published: November 2013

AbstractSeroprevalence ofBartonella henselae, Toxoplasma gondii, feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) infections was investigated in 1,447 domestic cats derived from the north (Hokkaido) to the south (Okinawa) prefectures in Japan. Of the cats investigated, 8.8% (128/1,447) were seropositive toB. henselae, 5.4% (78/1,447) toT. gondii, 9.8% (107/1,088) to FIV, and 2.9% (32/1,088) to FeLV, respectively. ForB. henselaeinfection, the positive rate varied from 11.5% in cats of 1 to <2 years old to 7.2% in those over 3 years old. Outdoor cats showed higher positive rate (14.5%) than that (7.0%) in indoor ones. The rate (13.5%) in flea‐infested cats was significantly higher than that (7.4%) in flea‐negative cats. The positive rates in southern and urban sites were more likely to be higher than those in northern and suburban sites, suggesting that warm and humid environments, density of cat population, and raising status, including hygienic condition and flea infestation in cats may correlate to higher seroprevalence ofB. henselaeinfection. ForT. gondii, FIV and FeLV infections, the seroprevalence also tended to be higher in outdoor, flea‐infested cats and advanced age groups. For FIV infection, the positive rates in male (14.3%) and outdoor cats (15.0%) were significantly higher than those in female (5.0%) and indoor cats (4.6%). On the other hand, no significant difference in seropositivities was observed in FeLV andT. gondiiinfections concerning to both genders and raising status.
Overcoming Tolerance in Hepatitis B Virus Transgenic Mice: A Possible Involvement of Regulatory T Cells
Authors: Roh Sujin , Kim Kilhyoun 
Published: November 2013

AbstractThe hepatitis B virus (HBV) transgenic mouse (Tg) 50–4 strain is immunologically tolerant to HBV antigens. Various vaccination strategies have been attempted but failed to break the tolerance in the mouse. Although the tolerance to HBV antigen is maintained, this mouse strain develops spontaneous liver disease beginning at the age of about 3 months. We attempted to induce immune responses to HBV surface antigen (HBsAg) in the Tg by immunization with recombinant vaccinia virus expressing HBsAg (vvHBV), and observed different immunological responsiveness between 2‐month‐old and 5‐month‐old Tg. In contrast to the unbreakable tolerance reported previously, we could induce both the cytotoxic T lymphocyte (CTL) and the antibody response against HBsAg by the vvHBV immunization. The cytokine expression pattern indicated that T helper 1 type immune response was induced. However, interestingly, these immune responses were observed only in the 5‐month‐old Tg, but not in the 2‐month‐old Tg. Furthermore, CD4+ T cells from 2‐month‐old mice, but not those from 5‐month‐old mice, inhibited CTL response to HBV antigen when adoptively transferred to C57BL/6. These results suggest the possible involvement of regulatory T cell function in the HBV Tg for maintaining tolerance. This study would contribute to a better understanding of immune status of the HBV Tg as a model of human chronic hepatitis and to the search for new therapeutic targets for chronic viral infections.
Induction of Apoptosis in Macrophage Cell Line, J774, by the Cell‐Free Supernatant from Pseudomonas aeruginosa
Authors: Zhang Jianling , Takayama Hisao , Matsuba Takashi , Jiang Ru , Tanaka Yoshinori 
Published: November 2013

AbstractPseudomonas aeruginosa is able to secrete many virulence factors that are cytotoxic towards eukaryotic cells. To investigate the effect of the bacterium on macrophages, we obtained cell‐free supernatants from P. aeruginosa (Pa) IID1117 (elastase‐positive and protease‐positive) and Pa IID1130 (elastase‐positive and protease‐negative). After 6 hr of incubation with the cell‐free supernatant from the Pa IID1117 strain, the viability of J774 macrophages was shown to be significantly reduced (47.5±11%), but not Pa IID1130 (96.4±1.6%) at a concentration of 10% (v/v) compared to control J774 macrophages without any supernatant (97.2±1.7%) by the detection of trypan blue dye exclusion. The death of cells was further demonstrated to be due to apoptosis characterized by chromatin condensation and apoptotic bodies by Hoechst 33258 staining, DNA fragmentation by agarose gel electrophoresis and terminal deoxynucleotidyl transferase‐mediated d‐UTP nick end labeling (TUNEL). An activated subunit was found to be released from procaspase‐3 in cell lysate. But in the presence of protease inhibitor, the apoptosis was completely blocked. The findings indicate that the Pa IID1117 strain is capable of inducing apoptosis in J774 macrophages. The apoptosis induced by the cell‐free supernatant from Pa IID1117 strain is suggested to be dependent on protease, but not elastase.
Effects of Tungstosilicate on Strains of Methicillin‐Resistant Staphylococcus aureus with Unique Resistant Mechanisms
Authors: Tajima Yutaka 
Published: November 2013

AbstractIn a previous study, it was found that polyoxotungstates such as undecatungstosilicate (SiW11) greatly sensitized strains of methicillin‐resistant Staphylococcus aureus (MRSA) to β‐lactams. In this report, the effects of SiW11 on several MRSA strains with unique resistant mechanisms were studied. SiW11 was still effective to MRSA mutants with higher β‐lactam resistance due to reduced cell‐lytic activity. Since the antimicrobial effect of TOC‐39 (a cephem antibiotic with strong affinity to penicillin‐binding protein (PBP) 2′) was not strongly enhanced in any case, it was confirmed that the sensitizing effect of SiW11 is due to reduced expression of PBP2′. However, the sensitizing effect of SiW11 was relatively weak in MRSA strains with lowered susceptibility to glycopeptide antibiotics. A certain resistant mechanism other than the mecA‐PBP2′ system worked in such a strain. Interestingly, an MRSA mutant with the Eagle‐type resistance was dramatically sensitized. This result suggests that SiW11 has another site of action besides reducing the expression of PBP2′.
Autologous Cancer Vaccine: A Novel Formulation
Authors: Ohno Tadao 
Published: November 2013

AbstractA novel cancer vaccine consisting of fixed autologous cancer tissue‐fragments, microparticles encapsulating cytokines, and an adjuvant was developed. In a Phase I/IIa clinical trial, vaccination to patients after resection of hepatocellular carcinoma induced significantly longer time before the first recurrence than that in historical control patients operated in the same department (P<0.05). This formulation will be promising against recurrence of many types of human cancers.
Comparative Studies of the Cell Structures of Mycobacterium leprae and M. tuberculosis Using the Electron Microscopy Freeze‐Substitution Technique
Authors: Takade Akemi , Umeda Akiko , Matsuoka Masanori , Yoshida Shin‐ichi , Nakamura Masahiro , Amako Kazunobu 
Published: November 2013

AbstractThe cell envelope and cytoplasmic architecture of the Mycobacterium leprae Thai‐53 strain were examined using the freeze‐substitution technique of electron microscopy and compared with those of the M. tuberculosis H37Rv strain. Both strains had similarly multilayered envelope architectures composed of an electron‐translucent layer, a peptidoglycan layer and the plasma membrane, from outside to inside. A comparison of the structures of these two mycobacteria revealed that the M. leprae cell was smaller in size and had a thinner peptidoglycan layer than the M. tuberculosis cell. The cell widths measured on electron micrographs were 0.44 μm for M. tuberculosis and 0.38 μm for M. leprae. The peptidoglycan layer of M. leprae was 4–5 nm, while the corresponding layer of M. tuberculosis was 10–15 nm.
Prevalence of Drug‐Resistant Human Immunodeficiency Virus Type 1 in Therapy‐Naive Patients and Usefulness of Genotype Testing
Authors: Ibe Shiro , Hotta Naoe , Takeo Uta , Tawada Yukio , Mamiya Naoto , Yamanaka Katsuo , Utsumi Makoto , Kaneda Tsuguhiro 
Published: November 2013

AbstractIn the present study, we performed genotypic drug‐resistance testing in 116 therapy‐naive human immunodeficiency virus type 1 (HIV‐1)‐infected patients between 1999 and 2002 at Nagoya National Hospital, Japan. The prevalence of drug‐resistant HIV‐1 with one or more major mutations significantly increased from 5.3% (4/75) in 1999–2001 to 17.1% (7/41) in 2002 (P=0.05), suggesting the spread of drug‐resistant HIV‐1. We identified a patient who possessed a protease (PR) inhibitor‐resistant HIV‐1 with a major mutation consisting of L90M before the initiation of therapy. The patient was administered zidovudine, lamivudine, and efavirenz as highly active antiretroviral therapy (HAART), as PR inhibitors were excluded based on the result of the drug‐resistance testing. The treatment succeeded in strongly suppressing the proliferation of drug‐resistant HIV‐1 and concomitantly increased CD4 cell counts. Thus, we conclude that drug‐resistance testing prior to the initiation of therapy is important for therapy‐naive patients to devise the optimum therapy regimen for each individual.