bluuebunny/crossref_metadata_2025_split huggingface dataset:
Crystallization of Synthetic Escherichia coli‐Type Lipid A
Authors: Kato Nobuo , Naito Setsuko , Arakawa Yoshichika , Sugiyama Tsuyoshi , Ito Hideo , Ohta Michio , Sasaki Kyoyu 
Published: November 2013

AbstractSynthetic Escherichia coli‐type lipid A formed hexagonal plate crystals when it was precipitated by the addition of 2 volumes of 95% ethanol containing 375 mM MgCl2 and incubated in 70% ethanol containing 250 mM MgCl2 at 4 C for 10 days. Analyses of crystals by electron diffraction and synchrotron X‐ray diffraction showed that crystals consist of hexagonal lattices with the lattice constant (a side of the lozenge as a unit cell on the basal plane) of 4.62 Å and the longitudinal axis (perpendicular to the basal plane) of 49.3 ±1.3 Å. Results suggest that the previous finding that various kinds of R‐form lipopolysaccharides crystallized but free lipid A isolated by acid hydrolysis from Re lipopolysaccharide did not crystallize under the same experimental conditions (Kato et al, J. Bacteriol., 172: 1516‐1528, 1990) is due to structural changes of lipid A occurring during the procedure of isolation of free lipid A.
Tumor‐Specific Transplantation Resistance in Mice after Treatment of Initial Tumors with Streptococcus thermophilus
Authors: Kaklij Gangadhar Supadu , Kelkar Shashikant Madan 
Published: November 2013

AbstractAntitumor activity observed by treatment with Streptococcus thermophilus was further investigated. The mice cured from fibrosarcoma by treatment with heat‐killed preparation of S. thermophilus, when challenged with fibrosarcoma failed to take up the tumor. However, these cured mice when challenged with sarcoma‐180 or Ehrlich ascites carcinoma, did not show significant changes in tumor take and/or survival compared to their respective controls. Similarly, mice cured from sarcoma‐180 were challenged with fibrosarcoma, sarcoma‐180 or Ehrlich ascites carcinoma. Though there was no change in the mean survival time (MST) of the dying mice regarding sarcoma‐180 or Ehrlich ascites carcinoma, there was 50 and 30% increase in the number of mice that showed total regression respectively over controls. However, there was no difference in the growth rate of fibrosarcoma. Similar observations were made with mice cured from Ehrlich ascites carcinoma, challenged with these tumors. These findings thus suggest that the antitumor response was tumor‐specific and that tumor‐associated antigens may have a role in imparting this specificity. Bacterial treatment non‐specifically augmented this primary response.
Adherence Patterns and DNA Probe Types of Escherichia coli Isolated from Diarrheal Patients in China
Authors: Xu Jian‐Guo , Cheng Bo‐Qun , Wu Yan‐Ping , Huang Li‐Bao , Lai Xin‐He , Liu Bing‐Yang , Lo Xing‐Zu , Li Hun‐Fen 
Published: November 2013

AbstractOne hundred and seventy‐two strains of Escherichia coli isolated from diarrheal patients in Beijing, P. R. China, were analyzed for plasmid DNA profile, HEp‐2 cell adherence ability and reactivity to 10 previously described DNA probes. They had not been recognized as pathogenic E. coli in China. Of the 110 strains tested, 76 (69%) contained one or multiple large plasmids. Of the 71 strains with the large plasmids 64 could adhere to HEp‐2 cells. Of the 172 strains, 102 (59.3%) were hybridized with at least one of the 10 probes. Of those, seven strains hybridized with enteroaggregative E. coli (EAggEC) probe. Their serotypes were O128 (two strains), O6 (one strain), and O111 (one strain). Three strains were untypable. Six and three strains were hybridized with enteropathogenic E. coli (EPEC) attaching and effacing genes (eae) or EPEC adherence factor (EAF) probe, respectively. Two non‐O157: H7 strains hybridized with enterohemorrhagic E. coli (EHEC) probe. Seventy‐two strains (41.9%) hybridized with shiga‐like toxin 2 or 1 (SLT2 or SLT1) probes. Among the SLT1 or SLT2 probe‐positive strains, 54 hybridized with invasive (INV) plasmid probe developed for identification of enteroinvasive E. coli (EIEC) and Shigella species. The INV and SLT probe‐positive strains might represent a new variety of verotoxin‐producing E. coli (VTEC).
Previous Enteric Adenovirus Infection Does Not Protect against Subsequent Symptomatic Infection: Longitudinal Follow‐up of Eight Infants
Authors: Unicomb Leanne E. , Jarecki‐Khan Kerstin , Hall Andrew , Podder Goutham 
Published: November 2013

AbstractEight infants followed longitudinally were found to have enteric adenovirus (EAdv) infections: in 5 infants with diarrhea and in 3 with no accompanying diarrhea. Sequential stool samples prior to EAdv infections were tested for adenovirus antigen, anti‐adenoviral IgA and neutralizing antibodies to serotypes 40, 41 and 2 in order to ascertain whether protection from symptoms was due to prior infection. No difference was found in the number of adenoviral infections among infants prior to their EAdv infections with or without accompanying diarrhea. However, in 3 of the 5 infants in whom EAdv infection was accompanied by diarrhea and 2 of 3 control infants, previous EAdv infections had occurred as detected by serotype‐specific antibody rises.
Murine Humoral Immune Response against Recombinant Structural Proteins of Hepatitis C Virus Distinct from Those of Patients
Authors: Ahmed Mamun , Shikata Toshio , Esumi Mariko 
Published: November 2013

AbstractWe examined the humoral immune response to recombinant structural proteins of hepatitis C virus (HCV) such as C, E1 and E2 in immunized mice. Mice showed high induction of antibodies against these three structural proteins. Conformational and/or linear epitopes of these regions showed high responses in mice. Comparison with patients revealed higher anti‐E1 and anti‐E2 responses in mice and 15 immunoreactive peptides which are unique to mice, especially 11 peptides from the E2 region. The hydrophilic regions of these proteins were found to be the most immunogenic. Therefore, the murine immune system against recombinant E1 and E2 glycoproteins was distinct from those of patients in natural infection, and may be a target to find protective activity against HCV infection.
Airway Interleukin‐8 in Elderly Patients with Bacterial Lower Respiratory Tract Infections
Authors: Ponglertnapagorn Preecha , Oishi Kazunori , Iwagaki Akitaka , Sonoda Fuminari , Watanabe Kiwao , Nagatake Tsuyoshi , Matsushima Kouji , Matsumoto Keizo 
Published: November 2013

AbstractWe investigated the interleukin‐8 (IL‐8) levels and neutrophil numbers in the sputum of 9 elderly patients with lower respiratory tract infections, including Pseudomonas aeruginosa infection, before and after treatment with various antimicrobial agents. The IL‐8 levels in sputum supernatants and the neutrophil numbers in sputum smears from 9 patients decreased significantly after the elimination of the causative respiratory pathogens. We also demonstrated that human recombinant IL‐8 at a range of 6.25‐25 ng/ml significantly enhanced opsonophagocytic killing of P. aeruginosa immunotype‐1 strain by human neutrophils in the presence of a serotype‐specific anti‐lipopolysaccharide monoclonal antibody and fresh normal human serum. These data suggest that the level of IL‐8 production in the airways of patients with lower respiratory tract infections is dependent on bacterial densities, and indicate the important role of IL‐8 not only in neutrophil migration but also in opsonophagocytic killing of bacteria in the lower respiratory tract.
Detection and Measurement of Staphylococcus epidermidis Slime Using an ELISA Technique
Authors: Mempel Martin , Günther Bernd , Müller Edgar , Grüter Ludwig 
Published: November 2013

AbstractA polyclonal rabbit anti‐serum against the strong slime‐producing Staphylococcus epidermidis strain RP62A was absorbed with the slime‐negative phase variant of this strain PV1 in order to remove not slime‐specific antibodies. Using this antiserum we established an ELISA which enables detection of slime production in S. epidermidis extracts. The ELISA showed high absorbance when extracts from slime‐positive strains (confirmed in the tissue culture tube test) were used as antigens. The high absorbance of slime‐positive strains was greatly reduced by periodate oxidation of the extracts and was resistant to proteinase digestion suggesting that the detected antigen is composed of polysaccharides. In contrast to other rapid and simple laboratory detection methods for S. epidermidis slime, the slime‐specific ELISA gave positive results in the presence of human serum.
Isolation of Coxiella burnetii from Children with Influenza‐Like Symptoms in Japan
Authors: Nagaoka Hiromi , Akiyama Masato , Sugieda Masaaki , Nishio Tomohiro , Akahane Sousuke , Hattori Hiroshi , Ho To , Fukushi Hideto , Hirai Katsuya 
Published: November 2013

AbstractThe prevalence of Coxiella burnetii antibodies was investigated by indirect immunofluorescence (IF) test in 55 paired sera (acute and convalescent phases) of school children who had influenza‐like symptoms. Of the convalescent serum samples examined, 18 (32.7%) sera reacted positively to phase II antigen of C. burnetii. Coxiella‐like organism was isolated from the sera of 13 children after injection of the 18 acute phase sera into mice. The organism was identified as C. burnetii by Giemsa staining and the IF antigen test of mouse spleen smears, the polymerase chain reaction (PCR) method, electron microscopic observations of the mouse spleen cells, and the IF antibody test of mouse sera. This is the first report of isolation of C. burnetii from serum specimens of children having influenza‐like symptoms. The evidence that C. burnetii was isolated from people indigenous to Japan at a considerably high incidence suggested that C. burnetii may be widespread as a cause of influenza‐like symptoms in Japan.
A Study of Two Procedures of HIV‐1 Isolation from Whole Blood Cultures
Authors: Shen Lu , Hober Didier , Benyoucef Samira , Ajana Faïza , Gérard Yann , Lion Georges , Vermersch Annie , Bocket‐Mouton Laurence , Mouton Yves , Wattré Pierre 
Published: November 2013

AbstractCulture techniques for isolation of HIV‐1 from small amounts of whole blood (WB) treated with anticoagulant have been reported and gave results identical to those of culture of separated peripheral blood mononuclear cells. Some authors obtained much higher isolation rates when EDTA was used instead of heparin. We compared two previously described techniques for cultivation of HIV‐1 from WB of adult HIV+ patients staged according to the CDC classification. In addition, we assessed the influence of the type of anticoagulant used for the collection of blood in viral replication in cell cultures from whole blood. Small volumes of WB treated with either heparin or EDTA were cocultivated with phytohemagglutinin (PHA)‐stimulated peripheral blood mononuclear cells (PHA‐PBMC) from healthy donors. We used two procedures for WB culture: procedure I, based on the culture of 250 μl of WB with 1 × 106 PHA‐PBMC from donors; and procedure II based on the culture of 500 μl of WB with 4× 106 PHA‐PBMC from donors. The cocultures were placed in 24‐well plates and incubated for as long as 28 days in medium containing interleukin 2 (IL‐2). Twice weekly half of the medium was replaced with fresh medium. In procedure II, one million fresh PHA‐PBMC from donors was added on the 7th day of culture. The culture supernatant was assayed for the presence of HIV‐1 p24 antigen in an enzyme immunoassay. The kinetics of HIV‐1 replication in cultures of WB from 7 AIDS patients were similar using procedures I and II. In 8 HIV + patients the isolation rate was higher with heparin‐ than with EDTA‐treated samples. The isolation rate was higher in AIDS patients (n = 8) than in others with both methods. In stage IV patients without AIDS (n = 8) we failed to isolate HIV‐1 in 1 patient with procedure I, whereas we succeeded with procedure II. In stage II, HIV‐I was isolated in 1 of 4 patients with both methods. HIV was isolated in cultures of WB from patients receiving zidovudine or related nucleoside analogues and in cultures of WB from untreated patients. HIV‐1 could not be isolated from WB of patients with more than 400 CD4+ T lymphocytes in their peripheral blood (n=4); however, it was isolated from 14 of 16 patients with less than 400 CD4+ T lymphocytes. Our results suggest that procedure II is more sensitive than procedure I and that heparin is better than EDTA for collecting WB. We showed that the rate of HIV‐1 isolation from WB increased in advanced‐stage patients. Further studies are needed to define the clinical applications of WB culture.
Detection and Sequencing of Norwalk‐Like Viruses from Stool Samples in Japan Using Reverse Transcription‐Polymerase Chain Reaction Amplification
Authors: Nakayama Miyuki , Ueda Yuhichi , Kawamoto Hiroyoshi , Han‐jun Yue , Saito Kunihiko , Nishio Osamu , Ushijima Hiroshi 
Published: November 2013

AbstractNorwalk‐like viruses were detected in Japan in 12% (26/209) of patients with nonbacteria and non‐rotavirus gastroenteritis in an outpatient clinic of a hospital from 1991 to 1994 by reverse transcription‐polymerase chain reaction. They were also present in 7% (26/378) of total samples including those from rotavirus‐positive gastroenteritis patients. In addition, the viruses were recovered in samples from 15 of 17 patients which were collected during outbreaks of gastroenteritis in various places in Japan by the same method. The DNA sequence of the polymerase region from patients at the hospital (sporadic cases) showed that two subgenogroups, similar to UK1‐6 in genogroup G1 and to UK1‐1 in genogroup G2 (Ando et al, J Clin Microbiol, 1995, 33: 64‐71) exist in Japan. The latter was more frequently found.
Relationship of HIV‐1 Envelope V2 and V3 Sequences of the Primary Isolates to the Viral Phenotype
Authors: Yoshimura Kazuhisa , Matsushita Shuzo , Hayashi Azusa , Takatsuki Kiyoshi 
Published: November 2013

AbstractWe examined the relationship between the amino acid sequences of the V2 and V3 regions of the envelope protein and the biological properties of ten human immunodeficiency virus type 1 (HIV‐1) primary isolates. The infectivity, cytopathic effect (CPE), and syncytium forming activity of these primary isolates were tested against three T cell lines (CEM, MT2, and MOLT4/CL.8 cells), CD8‐depleted peripheral blood mononuclear cells (PBMC), and primary monocyte‐derived macrophages (MDM) from seronegative donors. In addition to the viral groups which had the syncytium inducing/T‐cell line tropic (SI/TT) phenotype or non‐syncytium inducing/non‐T cell line tropic (NSI/NT) phenotype (including the NSI/macrophage tropic (NSI/MT) phenotype), there was a group of viruses that infected one or two T cell lines and PBMC but could not mediate syncytium formation. We therefore classified this group of viruses as a non‐syncytium inducing/partial T‐cell line tropic (NSI/pTT) virus. To investigate the relationship between these viral phenotypes and the sequence variability of the V2 and V3 regions of the envelope, we cloned the viral gene segment and sequenced the individual isolates. The sequence data suggested that the SI/TT type changes in the V3 sequence alone mediate a partial T cell line tropism and mild cytopathic effect and that an isolate became more virulent (SI/TT phenotype) if there were additional changes in the V2 or other regions. On the other hand, sequence changes in the V2 region alone could not mediate phenotypic changes but some additional changes in the other variable regions (for example, V3) might be required for the phenotypic changes in combination with changes in V2. These findings also suggested that amino acid changes in both the V2 and V3 region are required for the development of virulent variants of HIV‐1 that outgrow during advanced stages of the disease.
Distribution of Borrelia Species Associated with Lyme Disease in the Subalpine Forests of Nagano Prefecture, Japan
Authors: Nakao Minoru , Uchikawa Kimito , Dewa Hiroshi 
Published: November 2013

AbstractWe surveyed the natural distribution of Borrelia species associated with Lyme disease in the subalpine forests of Nagano prefecture, Japan, during 1993‐94. Tick‐derived isolates (n = 112) from Ixodes persulcatus and rodent‐derived isolates (n = 55) from Apodemus argenteus, Apodemus speciosus, Eothenomys andersoni, Eothenomys smithii, and Microtus montebelli were classified by rRNA gene restriction fragment length polymorphism analysis (RFLP ribotyping). Ribotype group IV (an intraspecific variant of Borrelia garinii) was predominant among the tick isolates. It was also isolated repeatedly from the rodents. Ribotype group III (Borrelia afzelii) was detected in low frequencies among the tick and rodent isolates. The data suggest that humans are likely to be exposed to the group IV when they are bitten by I. persulcatus ticks.
Monoclonal Antibody #5‐2‐26 Recognizes the Phosphatase‐Sensitive Epitope of Rabies Virus Nucleoprotein
Authors: Kawai Akihiko , Anzai Jun , Honda Yoshikazu , Morimoto Kinjiro , Takeuchi Kenji , Kohno Takashi , Wakisaka Koji , Goto Hideo , Minamoto Nobuyuki 
Published: November 2013

AbstractWe prepared monoclonal antibodies (MAbs) against the rabies virus N protein, among which one antibody (MAb 5‐2‐26) was shown to lack reactivity with the phosphatase‐treated N protein. The MAb was able to recognize the sodium dodecyl sulfate (SDS)‐denatured N protein. The MAb did not recognize the N‐protein analogues produced in Escherichia coli (E. coli), indicating that the N‐gene products were not normally processed in E. coli after translation. On the other hand, the MAb reacted normally with N‐gene products produced in COS‐7 cells, but not with those produced in the presence of K‐252a (a protein kinase inhibitor of a broad spectrum). The MAb displayed weak cross‐reactivity with the Triton‐insoluble network structures composed of several components, while another phosphoprotein (M1) of the virus was not recognized at all. These results suggest that MAb 5‐2‐26 preferentially recognizes a phosphatase‐sensitive linear epitope of N protein, which may enable further investigations to be conducted on the mechanism of N‐protein phosphorylation and its role(s) in virus replication.
A Monoclonal Antibody Modulates Neutrophil Adherence While Enhancing Cell Motility
Authors: Ohtake Kazuhisa , Takei Hiroshi , Watanabe Tadashi , Sato Yukiko , Yamashita Takao , Sudo Kazuhiko , Kuroki Motomu , Chihara Junichi , Sendo Fujiro 
Published: November 2013

AbstractA monoclonal antibody (MoAb) to human neutrophils, designated 3H9, was established by screening for the inhibition of neutrophil adherence to plastic plates containing a medium supplemented with fetal calf serum (FCS medium). The antigen recognized by 3H9 was shown to be present on human leukocytes and found at the highest levels on granulocytes. On Western blotting, 3H9 reacted with a molecule having a molecular weight of 80 kDa. When this MoAb was added at the same time as a neutrophil stimulant (fMLP), the inhibition of neutrophil adherence to plastic plates in the presence of FCS medium was observed after 60 min incubation. Furthermore, this MoAb enhanced not only fMLP‐induced chemotaxis but random migration of neutrophils as well. The mechanisms of these phenomena are discussed.
Nitrogen‐Gas Bubbling during the Cultivation of Clostridium tetani Produces a Higher Yield of Tetanus Toxin for the Preparation of Its Toxoid
Authors: De Luca María Marta , Abeiro Héctor Dario , Bernagozzi Jorge Alberto , Basualdo Juan Angel 
Published: November 2013

AbstractWe investigated the effect of exposing cultures of Clostridium tetani to nitrogen (N2) gas on the recovery of tetanus toxin to be processed for the preparation of its toxoid. N2 was bubbled through nine 10‐liter cultures during the growth of the bacteria, while nine parallel control incubations were maintained without bubbling. We found that treatment of the C. tetani anaerobes with an inert gas in this manner during cultivation produced a highly significant increase in the yield of tetanus toxin from them in comparison with the standard procedure.
In Vitro Amphotericin B Effects on Growth, Viability and Dimorphism of Paracoccidioides brasiliensis: Reversal of the Treatment
Authors: Jesuino Rosália Santos Amorim , de Bastos Ascenço Soares Renata , Salem‐Izacc Silvia Maria , Pereira Maristela , Felipe Maria Sueli Soares , de Almeida Soares Célia Maria 
Published: November 2013

AbstractThe in vitro effects of amphotericin B deoxycholate suspension (fungizone) on Paracoccidioides brasiliensis growth, cell viability and transformation were investigated. We also analyzed the protein synthesis patterns of both cellular forms, yeast and mycelium in the presence of AmB. This drug, at 30 μg/ml, highly inhibited yeast growth, which could be recovered depending on treatment time, where the most effective reversion was observed after 6 hr of incubation. The yeast cell viability, that had been partially affected by the drug, could also be efficiently recovered after AmB was removed. The effect of AmB on the cellular dimorphism process showed a strong reduction in the mycelium to yeast transformation (80% inhibition compared to the control without the drug). On the other hand, the transformation from yeast to mycelium in the presence of AmB was 50% affected, relative to the control. In contrast to the growth and cell viability experiments, the reversion effects on dimorphism were partial when the drug was removed, even with only 6 hr treatment. The two‐dimensional gels of 35S‐labeled proteins revealed a strong reduction in the three species of 80, 71 and 56 kDa in yeast and mycelium when treated with AmB.
Molecular Determination of Infection Source of a Sporadic Legionella Pneumonia Case Associated with a Hot Spring Bath
Authors: Miyamoto Hiroshi , Jitsurong Siroj , Shiota Ryoko , Maruta Koji , Yoshida Shin‐Ichi , Yabuuchi Eiko 
Published: November 2013

AbstractTo determine the infection source of a sporadic Legionella pneumonia case associated with a hot spring bath, we used five molecular methods, including repetitive element polymerase chain reaction (rep‐PCR), arbitrarily primed PCR (AP‐PCR), ribotyping, restriction endonuclease analysis (REA), and macrorestriction endonuclease analysis (MREA) by pulsed‐field gel electrophoresis. L. pneumophila serogroup (SG) 3 strain EY 3702, isolated from an intratracheal specimen of a 71‐year‐old Japanese female who developed pneumonia after nearly drowning in a hot spring spa bath, produced rep‐PCR and AP‐PCR fingerprints identical to those of L. pneumophila SG 3 strains EY 3768 and EY 3769 isolated from the bath water. Four epidemiologically unrelated L. pneumophila SG 3 strains showed different rep‐PCR or AP‐PCR fingerprints from those of the three EY strains (EY 3702, 3768, and 3769). The three EY strains were also genotypically indistinguishable by ribotyping with EcoRI and PstI, by REA with EcoBI or HindIII, and by MREA with NotI. Based on these results, we identified the bath water of the hot spring spa as the source of infection of this patient, even though the viable number of the organisms in the bath water was low (3 CFU/100 ml) when determined 27 days after her nearly drowning.
Detection of Flaviviruses by Reverse Transcriptase‐Polymerase Chain Reaction with the Universal Primer Set
Authors: Meiyu Fang , Huosheng Chen , Cuihua Chen , Xiaodong Tian , Lianhua Jiang , Yifei Peng , Weijun Chen , Huiyu Guo 
Published: November 2013

AbstractUsing a universal primer set designed to match the sequence of the NS1 gene of flaviviruses, the virus RNA of dengue (DEN), Japanese encephalitis (JEV), powassan and langat of Flaviviridae were successfully amplified by polymerase chain reaction (PCR) via cDNA; and with different internal primers, the serotypes of the dengue viruses were identified. Of the 78 clinically diagnosed dengue fever patients, 18 patients were positive for DEN 1, 48 patients for DEN 2 and 8 patients concurrently infected with DEN 4. Of the 52 patients admitted with Japanese encephalitis (JE), 45 were determined to be JEV infections. By nested PCR, we completed the identification of flaviviruses within 2 days. The results show that seven primers have a potential value for rapid clinical diagnosis of flavivirus infections.