bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of HIV‐1 Infection by an Intramolecular Antisense Peptide to T20 in gp160
Authors: Imai Masaki , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractAntisense amino acids are amino acids which can be translated from the corresponding anti‐codons of a sense amino acid. Antisense peptides encoded by the noncoding DNA strand have a tendency to interact with each other. We have demonstrated that antisense peptide sequences are present intramolecularly, and these may contribute to the folding and maintenance of the tertiary structure of a protein. T20 is a synthetic peptide with an amino acid sequence in the gp41 of HIV‐1 and has been demonstrated to be a potent inhibitor of HIV‐1 infection. We searched for intramolecular peptide sequences which are antisense to portions of T20. A synthetic peptide (TA‐1L) consisting of amino acids 84 to 97 of gp160, which contains an antisense peptide sequence (TA‐1) to T20, was shown to inhibit HIV‐1mB infection of MT‐4 cells. Interaction of these antisense peptides could be involved in sustaining HIV‐1 infectivity. The TA‐1L site, which exists in the C1 domain of gp160, is highly homologous among strains of HIV‐1, especially at TA‐1 and in the amino acids flanking the C terminus. Although the TA‐1 sites of 18 out of 30 HIV‐1 strains were antisense to the T20 region, those of the remaining 12 strains, including HIV‐1MN, were not. However, TA‐1L inhibited infection by HIV‐1MN, which has no antisense peptide in T20 corresponding to TA‐1, although the inhibitory effect was weaker. TA‐1L may thus also interfere with the gp160 interaction with CD4, which has an antisense sequence to TA‐1.
Cloning, Sequencing and Expression of the Gene Encoding the Extracellular Metalloprotease of Aeromonas caviae
Authors: Kawakami Kouji , Toma Claudia , Honma Yasuko 
Published: November 2013

AbstractA gene (apk) encoding the extracellular protease of Aeromonas caviae Ae6 has been cloned and sequenced. For cloning the gene, the DNA genomic library was screened using skim milk LB agar. One clone harboring plasmid pKK3 was selected for sequencing. Nucleotide sequencing of the 3.5 kb region of pKK3 revealed a single open reading frame (ORF) of 1,785 bp encoding 595 amino acids. The deduced polypeptide contained a putative 16‐amino acid signal peptide followed by a large propeptide. The N‐terminal amino acid sequence of purified recombinant protein (APK) was consistent with the DNA sequence. This result suggested a mature protein of 412 amino acids with a molecular mass of 44 kDa. However, the molecular mass of purified recombinant APK revealed 34 kDa by SDS‐PAGE, suggesting that further processing at the C‐terminal region took place. The 2 motifs of zinc binding sites deduced are highly conserved in the APK as well as in other zinc metalloproteases including Vibrio proteolyticus neutral protease, Emp V from Vibrio vulnificus, HA/P from Vibrio cholerae, and Pseudomonas aeruginosa elastase. Proteolytic activity was inhibited by EDTA, Zincov, 1,10‐phenanthroline and tetraethylenepentamine while unaffected by the other inhibitors tested. The protease showed maximum activity at pH 7.0 and was inactivated by heating at 80 C for 15 min. These results together suggest that APK belongs to the thermolysin family of metalloendopeptidases.
Constitutively Expressed phoP Inhibits Mouse‐Virulence of Salmonella typhimurium in an Spv‐Dependent Manner
Authors: Matsui Hidenori , Kawakami Takatoshi , Ishikawa Satoshi , Danbara Hirofumi , Gulig Paul A. 
Published: November 2013

AbstractIn Salmonella typhimurium, the transcription of several virulence genes including spvB is regulated by the PhoP/PhoQ regulatory system. To further examine the relationship between the PhoP/PhoQ and Spv systems for virulence in mice, we examined a non‐polar phoP mutation combined with different virulence plasmid genotypes for effects on virulence of S. typhimurium in the mouse model. PhoP–/Spv+ PhoP–/ Spv– mutants were not detectably recovered from the spleens of subcutaneously or orally inoculated mice. The phoP gene constitutively expressed from the lacZ promoter of a low copy number vector (phoPC) only partially complemented the non‐polar phoP mutation for mouse‐virulence in both the Spv+ Spv– backgrounds; both PhoPC strains exhibited virulence equal only to a PhoP+/Spv– strain. Interestingly, in a PhoP+ background, the phoPC gene reduced splenic infection of the Spv+ but not Spv– salmonellae after subcutaneous or oral inoculation compared with the PhoP+ parents. Additionally, the phoPC gene in an Spv+ background reduced the net growth of salmonellae in macrophages in vitro; phoPC in an Spv– background was without effect. These data suggest that the constitutive expression of the phoP gene attenuates the virulence of S. typhimurium in mice in an Spv‐dependent manner.
Clinical and Bacteriological Profiles of Patients with Typhoid Fever Treated during 1975–1998 in the Tokyo Metropolitan Komagome Hospital
Authors: Hoshino Yoshihiko , Masuda Gohta , Negishi Masayoshi , Ajisawa Atsushi , Imamura Akifumi , Hachimori Kei , Takayama Naohide , Yamaguchi Tsuyoshi , Kimura Mikio 
Published: November 2013

AbstractPatients with typhoid fever presenting to the Tokyo Metropolitan Komagome Hospital during the period 1975–1998 were retrospectively investigated. All cases were diagnosed by a positive culture for Salmonella typhi in either of their clinical specimens. Of the total number of 130 patients, 57% contracted the disease abroad; this population increased in later years as the total numbers of cases decreased. The period from disease onset to diagnosis averaged 14 days with 20% of the cases requiring over three weeks to establish a diagnosis. As for symptomatology relative bradycardia was seen in less than half of the cases, and rose spots or splenomegaly in less than one third. A positive blood culture was the most frequent test establishing the diagnosis followed by a positive stool culture. Intestinal bleeding was recognized in as many as 35 cases (27%) and even intestinal perforation occurred in two cases (1.5%). Chloramphenicol was most commonly employed during the early study period, however, during the late period it was replaced by fluoroquinolones. The clinical cure rate was 98% with regimens that include fluoroquinolones/quinolone; however it was 87% with the other antimicrobial regimens. Bacteriological relapse occurred in 25% of the non‐fluoroquinolone group while only in 2.0% in the fluoroquinolone/quinolone group. Four strains of Salmonella typhi that were multi‐resistant to chloramphenicol, ampicillin and cotrimoxazole were isolated in travelers from Asia. Early diagnosis by appropriate bacteriological examination regardless of classical symptomatology should be stressed and the use of fluoroquinolones is warranted in the treatment of typhoid fever.
Antibacterial Properties of Pseudomonas aeruginosa Immunotype 1 Lipopolysaccharide‐Specific Monoclonal Antibody (MAb) in a Murine Thigh Infection Model: Combined Effects of MAb and Ceftazidime
Authors: Akiyama Moritoshi , Oishi Kazunori , Tao Misao , Matsumoto Keizo , Pollack Matthew 
Published: November 2013

AbstractA murine monoclonal antibody (MAb) specific for the Pseudomonas aeruginosa immunotype 1 (It‐1) lipopolysaccharide (LPS) O‐side chain was evaluated in terms of its in vitro bactericidal opsonophagocytic activity and in vivo bacterial killing in a mouse thigh infection model. An immunoglobulin (Ig) G2a MAb Ld3–2F2, specific for It‐1 LPS, mediated in vitro complement‐dependent opsonophagocytic killing at a concentration of 10 μg/ml. MAb‐mediated, complement‐dependent killing also occurred in the absence of neutrophils at serum concentrations in excess of 20%. A remarkable synergy was observed in opsonophagocytic assays between MAb Ld3–2F2 (0.5 μg/ml) and ceftazidime (1/4 MIC). The administration of MAb Ld3–2F2 at a level of 1 μg resulted in a significant decrease in the number of bacteria in the thigh muscles of normal mice, while 100 μg of the same MAb was required for one log of reduction in the number of bacteria at the same site in neutropenic mice. The combined therapy with MAb Ld3–2F2 and ceftazidime provided a significant reduction in the density of bacteria in the thigh muscle at 9 hr post‐infection in normal and neutropenic mice as compared with those after treatment alone or with no treatment (P < 0.01). These favorable in vitro and in vivo interactions of an LPS‐specific IgG MAb and ceftazidime strongly support their potential for use in therapy, combined with an LPS‐reactive MAb and parenteral antipseudomonas β‐lactam antibiotics in the therapy of systemic Pseudomonas infections in normal and neutropenic hosts.
Effect of Ions on Antibacterial Activity of Human Beta Defensin 2
Authors: Tomita Tetsuji , Hitomi Shigemi , Nagase Takahide , Matsui Hirotoshi , Matsuse Takeshi , Kimura Satoshi , Ouchi Yasuyoshi 
Published: November 2013

AbstractHuman beta defensin 2 (HBD‐2), the most recently discovered human defensin, has been considered to work as a host defense substance against microbial infection. Using Escherichia coli ATCC 25922, we investigated how some cations and anions influenced the antimicrobial activity of HBD‐2. This activity, measured in 10 mm phosphate buffer at a concentration of 20 μg/ml, reduced significantly in the presence of 100 and 150 mm sodium or potassium chloride. The reduction was not significantly different when the total amounts of sodium and potassium ions were equal. The kind and the valence of anions (chlorine and sulfate ions) did not affect the bactericidal activity as long as the concentrations of sodium ions were equal. Divalent ions (calcium and magnesium ions) added to 10 mm of Tris buffer significantly inactivated HBD‐2 at much lower concentrations (more than or equal to 0.01 mm and 0.05 mm, respectively) than the monovalent ions did. These findings suggest that HBD‐2 kills the bacteria through at least two phases, which are affected independently by either monovalent or divalent ions and unaffected by anions.
Mechanism of High Susceptibility of Iron‐Overloaded Mouse to Vibrio vulnificus Infection
Authors: Hor Lien‐I , Chang Yir‐kun , Chang Chien‐chung , Lei Huan‐yao , Ou Jonathan T. 
Published: November 2013

AbstractVibrio vulnificus produces fulminant septicemia in humans with underlying conditions, particularly those with diseases that elevate the iron level. The effect of a high iron level on the virulence of V. vulnificus was therefore investigated in mice treated with iron dextran. The mice loaded with iron became highly susceptible to V. vulnificus infection, the LD50 (50% lethal dose) decreased five logs when infected per peritoneum. However, when infected via the oral route, the LD50 was affected little unless the mouse was treated with an additional drug such as cyclophosphamide or d‐galactosamine. Mice with or without iron‐overloading died when the bacterial concentration in the blood reached 105 cfu/ml or above. Iron increased the growth rate of the bacteria, both inside and outside of the animal, quickly reaching a lethal concentration in the iron‐overloaded mouse. V. vulnificus, grown with or without the addition of iron, showed strong cytotoxicity on the isolated cells or within the animal at high bacterial concentration. Iron overload stimulated the production of tumor necrosis factor a (TNF‐α), a major factor of septic shock, in mice upon infection with the bacteria, probably caused by the endotoxin; however, the neutrophils, whose migration is effected by TNF‐α, appeared to be less active. Taken together, the major virulence factor of V. vulnificus appeared to be the accelerated growth of bacteria to quickly reach the lethal level and the lower activity of immune cells including neutrophil as a result of iron‐overloading. These two effects manifest other virulence factors, the host's as well as bacterial. Such factors, other than TNF‐α stimulated by the endotoxin, enhanced cytotoxicity, which kills the host cells including the host's immune cells.
Serological Properties of Abiotrophia and Granulicatella Species (Nutritionally Variant Streptococci)
Authors: Kitada Katsuhiro , Okada Yasuko , Kanamoto Taisei , Inoue Masakazu 
Published: November 2013

AbstractSerological variations were examined among 12 type or reference strains and 91 oral isolates of vitamin B6‐dependent Abiotrophia and Granulicatella spp. Rabbits were immunized with whole cells of 12 selected strains and 10 typing antisera were obtained, which were unreactive with the Lancefield group A to G antigen preparations. The reactivity of the antisera and autoclaved cell surface antigen extracts was tested by double diffusion in agar gel and a capillary precipitin test. These typing antisera categorized all Abiotrophia defectiva strains, all except one Granulicatella elegans strain, three‐quarters of the Granulicatella adiacens, and half of the Granulicatella paraadiacens into 8 serotypes and 2 subserotypes. The Granulicatella balaenopterae type strain was unserotypable. All A. defectiva strains were serotype I, some of which were divided into subserotype I‐1 and/or I‐5. The G. adiacens strains generally belonged to serotype II or III, and the G. paraadiacens strains to serotype IV, V or VI. All G. adiacens or G. paraadiacens serotype II strains were also subserotype I‐5. The G. elegans strains were serotype VII or VIII. These Abiotrophia and Granulicatella serotypes were undetectable among 33 strains of the other 11 species including the bacteriolytic enzyme‐producing but vitamin B6‐independent strains of Streptococcus, Enterococcus, Dolosigranulum and Aerococcus. The proposed serotyping system for Abiotrophia and Granulicatella spp. would be helpful in the identification and classification of these unique coccal isolates in ecological and epidemiological studies.
Acute Primary Infection with Cytomegalovirus (CMV) in Kidney Transplant Recipients Results in the Appearance of a Phenotypically Aberrant CD8+ T Cell Population
Authors: Van Dam Joanne G. , Damoiseaux Jan G.M.C. , Christiaans Maarten H.L. , Bruggeman Cathrien A. 
Published: November 2013

AbstractHuman cytomegalovirus (CMV) is a β‐herpesvirus that causes a chronic subclinical infection in healthy man. The immune system is unable to eliminate the virus completely, allowing virus to persist in a latent state. In the immunocompromised host, this equilibrium is disturbed, resulting in a clinical infection. In immunocompromised rats, clinical CMV infection is associated with an increase in NK cells and CD8+ T cells, including a phenotypically aberrant CD8+ T cell population. Using flow cytometry, we examined the effect of acute CMV infection on the composition of leukocyte subsets in immunocompromised patients. Therefore, we used peripheral blood of CMV seronegative patients receiving a kidney from a seronegative (control group) or a seropositive donor. Of the patients receiving a seropositive kidney, only the patients undergoing acute CMV infection were included (experimental group). Special attention was paid to the phenotype of the cytotoxic T cells. The development of acute CMV infection resulted in an increased NK cell number and an activation of both CD4+ and CD8+ T cells, as determined by HLA‐DR expression. An aberrant CD8+ T cell subset with decreased expression of CD8 and TCRαβ appeared in the infected patients. Furthermore, the size of this subpopulation of CD8+ T cells is positively correlated with the viral load.
Seroprevalence of Antibodies against Spotted Fever Group Rickettsia among Dogs and Humans in Okinawa, Japan
Authors: Satoh Hiroshi , Tsuneki Akemi , Inokuma Hisashi , Kumazawa Norichika , Jahana Yoshihide , Kiyuuna Tsuyoshi , Okabayashi Tamaki , Muramatsu Yasukazu , Ueno Hiroshi , Morita Chiharu 
Published: November 2013

AbstractWe evaluated serum antibodies against Rickettsia japonica in 517 dogs (430 stray dogs and 87 pet dogs) and 164 humans in Okinawa, Japan, by indirect immunofluorescence assay. The seropositive rate in stray dogs was significantly higher than that in pet dogs (30.7 versus 4.6%, P<0.01). This high prevalence rate is attributed to the understandably frequent environmental exposure of stray dogs to tick infestation. Human samples obtained from Okinawa and Sapporo also showed a significant difference in seropositive antibody percentages (45.1 and 12.0%, respectively, P<0.01). This result suggests that there has been preexposure to spotted fever group rickettsia in humans in Okinawa.
Serological Analysis of Human Leptospirosis in the Philippines
Authors: Masuzawa Toshiyuki , Dancel Louella A. , Miyake Masaki , Yanagihara Yasutake 
Published: November 2013

AbstractLeptospirosis in the Philippines is an underrepresented disease. To achieve an accurate means of serodiagnosis, we demonstrated antibodies to the prevalent Leptospira serovars in sera of 71 patients from three major hospitals in Manila by the microscopic agglutination test and Western blot analysis. Sera of 53 patients contained antibody against 8 serovars poi, tarassovi, manilae, pyrogenes, australis, grippotyphosa, javanica, and autumnalis.
Molecular Characterization in the VP7, VP4 and NSP4 Genes of Human Rotavirus Serotype 4 (G4) Isolated in Japan and Kenya
Authors: Kudo Satoshi , Zhou Yumei , Cao Xin‐Ru , Yamanishi Shigeki , Nakata Shuji , Ushijima Hiroshi 
Published: November 2013

AbstractThe VP7, VP4 and NSP4 genes of human rotavirus serotype 4 (G4) were analyzed to investigate intraserotypic variations. The techniques used included reverse transcription polymerase chain reaction with subtype specific primers, restriction fragment length polymorphism analysis and sequence analysis. Twelve isolates (nine from Japan and three from Kenya) and two standard strains (Hochi, Odelia) were G4A P[8] Wa group NSP4. A standard strain (ST3) was G4A P[6] Wa group NSP4 and a strain (VA70) was G4B P[8] Wa group NSP4. These results show G4 rotaviruses can be divided into three combinations at the moment.
Iron‐Mediated Regulation of Alkaline Proteinase Production in Pseudomonas aeruginosa
Authors: Shigematsu Takashi , Fukushima Jun , Oyama Masatsune , Tsuda Masataka , Kawamoto Susumu , Okuda Kenji 
Published: November 2013

AbstractWe analyzed the regulation by iron of alkaline proteinase (AP) production in Pseudomonas aeruginosa. Extracellular AP production was detected from the mid‐logarithmic to the stationary phase by an antibody‐based assay system, and was strongly repressed by iron in the medium. This repression was shown by Northern hybridization and primer extension to occur at the level of transcription. The primer extension analysis revealed that the start point of transcription of AP gene was the nucleotide position −84 from the start point of translation. Furthermore, we investigated whether this transcriptional repression involved PvdS protein. Using the mutant strain of pvdS, the alternative σ factor gene revealed that the PvdS protein is required for the full expression of AP, and a previous study showed that expression of pvdS is also repressed by iron. Therefore, we thought that one mechanism of repression of AP production operated through reduction of the PvdS protein level. Purified AP decomposed the transferrin, and released iron from it Purified AP added to the medium containing transferrin as the only iron source enhanced the growth of P. aeruginosa. Moreover, mutation in the AP gene decreased the growth rate in the medium containing the transferrin as the only iron source. These results clearly indicated that AP expression should occur in a free‐iron‐deficient environment and emphasized the importance of AP to iron acquisition in the infection site.
A Rapid Bioluminescent Enzyme Immunoassay (BLEIA) for the Detection of Shiga Toxin Types 1 and 2
Authors: Yamazaki Makoto , Sato Sachihiro , Gondaira Fumio , Sugiyama Jun‐ichi 
Published: November 2013

AbstractIn recent years, Escherichia coli O157: H7 has emerged as a global public health concern. Among the more important virulence characteristics of this strain is its ability to produce one or more Shiga toxins (Stx). Traditional culture‐based methods for assay of enteric toxins in foods and clinical samples are relatively slow and results can be ambiguous. In this work, we established a toxin‐detection system based on bioluminescent enzyme immunoassay (BLEIA) using a simple and inexpensive device. The system could detect both Shiga toxin types 1 and 2 individually within 150 min with a detection limit for each toxin at 5 pg/ml. In our study of previously characterized Shigatoxigenic and all non‐Shigatoxigenic E. coli and other bacterial species, we found all Shigatoxigenic strains to be positive and non‐Shigatoxigenic E. coli and other bacterial species to be negative. This assay was also used to detect Stxs in milk and supernatant fluids from minced chicken and beef. For clinical stool samples we noted a tendency for the system to give unexpectedly high background level. Our results suggest the feasibility of using BLEIA methodology for the simple, rapid and sensitive detection of toxins from culture supernatant, various foods and clinical samples.
Paramyxovirus Accessory Proteins as Interferon Antagonists
Authors: Gotoh Bin , Komatsu Takayuki , Takeuchi Kenji , Yokoo Junko 
Published: November 2013

AbstractA new role of the Paramyxovirus accessory proteins has been uncovered. The P gene of the subfamily Paramyxovirinae encodes accessory proteins including the V and/or C protein by means of pseudotemplated nucleotide addition (RNA editing) or by overlapping open reading frame. The Respirovirus (Sendai virus and human parainfluenza virus (hPIV)3) and Rubulavirus (simian virus (SV)5, SV41, mumps virus and hPIV2) circumvent the interferon (IFN) response by inhibiting IFN signaling. The responsible genes were mapped to the C gene for SeV and the V gene for rubulaviruses. On the other hand, wild type measles viruses isolated from clinical specimens suppress production of IFN, although responsible viral factors remain to be identified. Both human and bovine respiratory syncytial viruses (RSVs) counteract the antiviral effect of IFN with inhibiting neither IFN signaling nor IFN production. Bovine RSV NS1 and NS2 proteins cooperatively antagonize the antiviral effect of IFN. Studies on the molecular mechanism by which viruses circumvent the host IFN response will not only illustrate co‐evolution of virus strategies of immune evasion but also provide basic information useful for engineering novel antiviral drugs as well as recombinant live vaccine.
Fungal Flora on Board the Mir‐Space Station, Identification by Morphological Features and Ribosomal DNA Sequences
Authors: Makimura Koichi , Hanazawa Ryo , Takatori Kosuke , Tamura Yoshiko , Fujisaki Ryuichi , Nishiyama Yayoi , Abe Shigeru , Uchida Katsuhisa , Kawamura Yoshiaki , Ezaki Takayuki , Yamaguchi Hideyo 
Published: November 2013

AbstractThis report is on the morphological and molecular biological identification, using 18S‐ and ITS1‐rDNA sequences, of the “space fungi” isolated on board the Russian Mir‐Space Station as the major constituents of the fungal flora. The six fungal strains were isolated from air by using an air sampler or from condensation. Strains were identified as Penicillium chrysogenum, Aspergillus versicolor, or Penicillium sp. by both methods. The species of space fungi were common saprophytic fungi in our living environment, potential pathogens, and allergens. This study concluded that the environment on board the space station Mir allows the growth of potentially pathogenic fungi as true in residential areas on the earth. Therefore, to prevent infection or other health disorders caused by these fungi, easy and reliable methods should be established to survey the fungal flora in a space station.