bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative Analysis of Polymyxin B Released from Polymyxin B‐Treated Dormant Spores of Bacillus subtilis and Relationship between Its Permeability and Inhibitory Effect on Outgrowth
Authors: Fujita‐Ichikawa Yuko , Tochikubo Kunio 
Published: November 2013

AbstractPolymyxin B, one of the cyclic polypeptide antibiotics, binds to the coat of Bacillus subtilis dormant spores and inhibits them from growing after germination. When about 2.8 × 108 cells/ml of polymyxin B‐treated dormant spores were incubated in heart infusion broth, 3.6 μg/ml of polymyxin B were released into the liquid medium during germination. Incubation of the same concentration of polymyxin B‐treated ones in 100 mM CaCl2 solution released 4.0 μg/ml of the antibiotic. The effect of various concentrations of polymyxin B on germination, outgrowth and vegetative growth of the dormant spores was investigated; the results showed that concentrations of 4.0 μg/ml and higher of the antibiotic inhibited their outgrowth and vegetative growth after germination. Young vegetative cells were less sensitive to the antibiotic than germinated spores. In addition to these results, immunoelectron microscopy with colloidal gold particles indicated that polymyxin B permeated into the core of the germinated spores and inhibited them from outgrowing.
Antigen‐Specific Proliferative Response of Peritoneal Exudate Lymphocytes Primed with Antigen and Bacterial Lipopolysaccharide: The Roles of Ia+ Accessory Cells and IL‐2
Authors: Nitta Toshimasa , Imai Hiroaki , Ogasawara Yuko , Hirayama Nobukuni , Nakano Masayasu 
Published: November 2013

AbstractIn vitro antigen‐specific proliferation was investigated in a lymphocyte population that had been taken from the peritoneal exudate cells (PEC) of C3H/HeN mice (Iak) primed in vivo with both bacterial lipopolysaccharide (LPS) and horse red blood cells (HRBC) and had been purified by passage through a nylon fiber column (Nfc). The proliferative response of the Nfc‐passed lymphocytes primed with HRBC and LPS [T(HRBC + LPS) cells] depended on the dose of antigen in the cultures, and the response was higher than that of cells prepared from mice primed with HRBC alone [T(HRBC) cells]. No response was seen in the cells prepared from the LPS‐primed mice [T(LPS) cells] or normal mice [T(N) cells]. The response of the T(HRBC) cells was abolished by previous treatment of the cells with anti‐Iak antibody and complement (C), whereas the response of the T(HRBC + LPS) cells was retained after the same treatment, indicating that the Ia– T(HRBC + LPS) cells can proliferate in response to antigen in spite of Ia+ accessory cell‐depletion. Supernatants from the cultures of Ia– T(HRBC + LPS) cells in the presence of HRBC showed abundant IL‐2 activity, while those of Ia– T(HRBC) cells did not. The IL‐2 should be produced by the L3T4 cell population in T(HRBC + LPS) cells in response to antigen, since the previous treatment of the cells with anti‐L3T4 antibody and C abrogated the production. On the other hand, the Ia– T(HRBC + LPS) cells as well as the Ia– T(LPS) cells could respond to IL‐2 dose‐dependently when recombinant IL‐2 was added into the cultures, but the response of Ia– T(HRBC) cells to IL‐2 was very weak. The cell population responding to IL‐2 in the T(HRBC + LPS) cells as well as T(LPS) cells must be AsGM1‐positive or natural killer (NK) cells, since previous treatment of the cells with anti‐AsGM1 antibody and C abrogates the response. Together these results suggest that L3T4 lymphocytes capable of producing IL‐2 in response to HRBC antigen without Ia+ accessory cells are generated in the PEC of the mice after priming with LPS and antigen together, and the IL‐2 produced by the L3T4 lymphocytes induces the proliferation of the LPS‐primed AsGM1+ cells.
Characterization of Cell Growth‐Inhibitory Factor in Inflammatory Peritoneal Exudate Cells of Rats
Authors: Yui Satoru , Yang De , Mikami Masaaki , Yamazaki Masatoshi 
Published: November 2013

AbstractWe characterized the nature and reaction mode of the cell growth‐inhibitory factor (here designated CGIF) from rat peritoneal exudate cells (PEC). The soluble fraction separated from the lysate of Enterococcus faecalis‐induced 24 hr PEC completely inhibited Con A‐induced thymocyte mitogenesis. Gel filtration chromatography showed that CGIF has a molecular weight of approximately 23–25 kDa. Isoelectric focusing with Rotofor indicates that the factor has an isoelectronic point of 5.8–6.4. CGIF was inactivated by treatment at 70 C, for 30 min or by tryptic digestion, but the activity was not destroyed by the reduction with dithiothreitol. As well as thymocyte proliferation, CGIF completely suppressed 3H‐thymidine incorporation of splenocytes which were stimulated by either Con A or LPS, suggesting the factor is effective on both T and B cells. The acting point of the inhibitor appeared to be a later stage of the lymphocyte activation sequence, since it was still effective when added 28.5 hr after the addition of Con A. CGIF also reduced the viability of these cells when added with mitogens such as Con A or LPS. CGIF thus appears to be distinct from interleukin‐1 receptor antagonist or transforming growth factor‐β.
Diagnosis of Genital Herpes by Polymerase Chain Reaction Amplification
Authors: Goto Tetsuya , Yamaguchi Yuki , Hashido Madoka , Yoshikawa Hiroyuki , Kawana Takashi 
Published: November 2013

AbstractA new polymerase chain reaction (PCR) method employing type‐specific primers and probes was applied to 114 clinical specimens obtained from 58 female patients with genital lesions or who had a history of genital herpes. Ten and 15 specimens, respectively, were positive for herpes simplex virus (HSV)‐1 and HSV‐2 by cell culture. All of 10 culture‐confirmed HSV‐1 cases and 11 of 15 (73%) culture‐confirmed HSV‐2 cases were identified by PCR. Although there were several cases with discrepancy between cell culture and PCR for HSV‐2, the results suggest that this PCR procedure could be applied to clinical specimens from the female genital tract.
Efficient Adsorption of Lysostaphin on Bacterial Cells of Lysostaphin‐Resistant Staphylococcus aureus Mutant
Authors: Sakurada Junji , Murai Miyo , Zhijun Li , Usui Akemi , Seki Keiko , Kobayashi Kiyoshi , Sumi Yoshiko , Jitsukawa Hiroko , Masuda Shogo 
Published: November 2013

AbstractA simple and efficient method for the purification of staphylolytic endopeptidase (lysostaphin) contained in culture supernatant of Staphylococcus simulans biovar staphylolyticus strain by adsorption of the enzyme on bacterial cells of lysostaphin‐resistant S. aureus mutant was successfully devised. Lysostaphin was sufficiently adsorbed on the heat‐killed mutant cells derived from S. aureus Cowan I and efficiently eluted by 3 M KSCN. Enzyme preparation obtained by a single procedure of the affinity purification was pure enough for practical use. The yield of the enzyme was 25 mg from 1 liter culture and recovery rate was 64%.
Construction of a Slime Negative Transposon Mutant in Staphylococcus epidermidis Using the Enterococcus faecalis Transposon Tn917
Authors: Grüter Ludwig , Feucht Hubert , Mempel Martin , Laufs Rainer 
Published: November 2013

AbstractThe slime‐producing Staphylococcus epidermidis strain sensu strictu CNS23 was transformed by protoplast transformation with the plasmid pTV1 which carries transposon Tn917. Using this transposon mutagenesis system we obtained the Tn917‐inserted mutant CT512, which has lost the ability to produce slime. A single insertion of the trasposon Tn917 into the chromosome of CT512 could be detected by Southern hybridization. This mutant showed a significantly higher stability concerning its slime‐negative phenotype compared with spontaneous slime‐negative mutants of S. epidermidis strain CNS23. In slime‐ELISA no slime‐associated antigen could be detected in extracts of the transposon mutant. Compared to slime‐positive S. epidermidis strains, CT512 lacked in accumulative growth in microtiter tube test.
Suppression of ADCC by Immune Complexes Formed in Vitro in Mycobacterium leprae‐Infected Mice
Authors: Vaishnavi Chetana , Ganguly Nirmal Kumar , Kaur Surrinder , Kumar Bhushan 
Published: November 2013

AbstractAntibody‐dependent cellular cytotoxicity (ADCC) was assessed in mice infected experimentally with Mycobacterium leprae and injected simultaneously with in vitro‐formed immune complexes (IC). Significant decrease in the ADCC function was observed in animals given IC at zero day (OdIC) and 3 months (3mIC) post inoculation with M. leprae, when ADCC activity was assessed at 3, 6 and 9 months period. From the data obtained we believe that ADCC is suppressed by IC formed in vitro.
Inhibitory Effect of 8‐(N, N‐Diethylamino)octyl‐3, 4, 5‐Trimethoxybenzoate Hydrochloride (TMB‐8) on Germination of Bacillus cereus T Spores
Authors: Shibata Hirofumi , Mimura Akiko , Mitani Hirotoshi , Tani Isamu , Hashimoto Tadayo 
Published: November 2013

AbstractEffect of 8‐(N,N‐diethylamino)octyl‐3, 4, 5 ‐ trimethoxybenzoate hydrochloride (TMB‐8), a calcium antagonist, on germination of Bacillus cereus T spores induced by L‐alanine and inosine was investigated. TMB‐8 had no effect on the germination of heat‐activated spores, whereas it inhibited that of nonactivated spores. The TMB‐8 inhibitory effect was antagonized competitively by inosine, but not by L‐alanine. Addition of Ca2+ reversed the inhibitory effect of TMB‐8 in a dose‐related fashion. Based on the results, a role of inosine and a site(s) for inhibitory action of TMB‐8 in the process leading to the germination of nonactivated spores were discussed.
Protective Activity of Antisera against Isolates of Borrelia burgdorferi from Various Geographical Origins
Authors: Masuzawa Toshiyuki , Okada Yutaka , Yanagihara Yasutake 
Published: November 2013

AbstractAntisera from rabbits immunized with two Japanese strains of Borrelia burgdorferi, HP3 an isolate from Ixodes persulcatus and HO14 an isolate from I. ovatus, or the European strain P/Bi isolated from human cerebrospinal fluid (CSF) did not passively protect hamsters from challenge with the infectious strain 297, a North American isolate from patient CSF. Antisera to strains 297 and B31, a North American isolate from I. dammini, however, provided protective effect to challenge with strain 297. Immune mice sera in the presence of homologous B. burgdorferi antigen induced the production of oxygen intermediates from mouse peritoneal exudate cells. Heterologous B. burgdorferi antigen had no effect. These results suggest that antigenic properties of Japanese strains are different from those of North American and European isolates.
Identification of a Human Cancer Related Organ‐Specific Neoantigen
Authors: Ilantzis Christian , Thomson David M.P. , Michaelidou Alice , Benchimol Sarita , Stanners Clifford P. 
Published: November 2013

AbstractHuman cancers express organ‐specific neoantigens (OSNs) which elicit specific cellular immune responses in the cancer patient, as demonstrated by leukocyte adherence inhibition (LAI), an in vitro immune response assay. A purified protein of MW 40,000 (p40) exhibiting OSN (colon specific) activity was cleaved into specific peptide fragments and their partial amino acid sequences determined. This information was used in the polymerase chain reaction (PCR) to obtain a 992 bp cDNA clone (PCR‐992) from a human colon adenocarcinoma cell line (LS‐180). By comparison of the predicted amino acid sequence of PCR‐992 with the known sequence of p40 peptides, PCR‐992 was shown to correspond to almost the entire coding region of p40. Nucleotide sequence analysis suggested that the protein was mycoplasmal in origin due to its high A+T content (76%) and the presence of five in frame TGA termination codons; at least two of the latter are actually read as tryptophan, a known feature of mycoplasma translation. We have confirmed this origin by direct isolation of a contaminating mycoplasma species from the LS‐180 cell line and demonstration that it could be hybridized with the PCR‐992 probe. Northern and PCR analysis of RNA preparations from the contaminated LS‐180 cell line showed that p40 was part of the high affinity transport system operon of Mycoplasma hyorhinis (Dudler et al, EMBO J., 7: 3963‐3970, 1988). Total protein lysates of Mycoplasma hyorhinis cultivated without animal cells could elicit positive LAI responses when incubated with cancer patient leukocytes but not with normal patient leukocytes. The organ‐specific nature of the response was, however, not observed indicating that host cell‐mycoplasmal interactions may play a role in determining the organ‐specific nature of p40 seen with the LAI. The significance of these findings will be discussed in the context of previous thinking regarding the origin of OSNs.
Collagen‐Induced Arthritis Suppressed with Monoclonal Anti‐Idiotypic Antibody
Authors: Tarutani Susumu 
Published: November 2013

AbstractIn foregoing work, we identified at least 5 distinct epitopes on human type II collagen (CII), using 8 murine monoclonal antibodies (mAb) against human CII, and suggested that a species‐nonspecific epitope on CII recognized by anti‐CII mAb termed 1‐5 is an arthritogenic epitope. We also found that antibody response against a selected epitope of human CII could be induced by immunization with rabbit anti‐idiotypic (Id) antibody against anti‐CII mAb. The author developed and characterized monoclonal anti‐Id antibodies against 1‐5 mAb recognizing a putative arthritogenic epitope. The author also investigated whether the anti‐Id mAb could regulate antibody response directed against a selected epitope recognized by 1‐5 mAb, and the induction of collagen‐induced arthritis in DBA/1J mice. DBA/1J mice intravenously preinjected with anti‐Id mAb to 1‐5, did not produce anti‐CII antibody expressing 1‐5 Id upon immunization with human CII. Furthermore, as the development of collagen‐induced arthritis (CIA) in DBA/1J mice pretreated with anti‐Id mAb to 1‐5 was significantly suppressed, anti‐Id mAb will be a useful tool for studying the regulation of antibody response to a selected epitope. This study lends support to our hypothesis that the 1‐5 epitope is an arthritogenic epitope.
Lipopolysaccharide Isolated from a New O‐Antigenic Form (O13) of Vibrio parahaemolyticus
Authors: Hisatsune Kazuhito , Iguchi Takehiro , Haishima Yuji , Tamura Norihiko , Kondo Seiichi 
Published: November 2013

AbstractThe chemical properties of a lipopolysaccharide (LPS) isolated from a new O‐antigenic form (O13) of Vibrio parahaemolyticus were investigated. The LPS contained glucose, galactose, L‐glycero‐D‐manno‐heptose and glucosamine. 2‐Keto‐3‐deoxy‐octonate (KDO) was not detected in the LPS by the periodate‐thiobarbituric acid test (Weissbach's reaction) under conventional hydrolysis conditions. Instead, phosphorylated KDO (X1 and X2) was found in its strong‐acid hydrolysate. This sugar composition was identical to that of V. parahaemolyticus O3, O5 and O11 LPS, indicating that, based on the sugar composition, O13 LPS belongs to Chemotype III to which O3, O5 and O11 belong. In addition, structural study demonstrated the presence of KDO 4‐phosphate in its inner‐core region.
Identification of Heat‐Stable Enterotoxin‐Producing Strains of Yersinia enterocolitica and Vibrio cholerae Non‐O1 by a Monoclonal Antibody‐Based Enzyme‐Linked Immunosorbent Assay
Authors: Nair G. Balakrish , Bhattacharya Sujit K. , Takeda Tae 
Published: November 2013

AbstractUsing a mouse monoclonal antibody (MAb) 2F raised against Vibrio cholerae non‐O1 heat‐stable enterotoxin (NAG‐ST) which also recognizes a shared epitope of Yersinia enterocolitica heat‐stable enterotoxin (Y‐ST), a competitive enzyme‐linked immunosorbent assay (ELISA) was developed for independent detection of NAG‐ST and Y‐ST. There was good concordance between the Y‐ST ELISA and the suckling mouse assay (SMA) for detection of Y‐ST from test strains of Y. enterocolitica, and the Y‐ST ELISA can effectively replace the SMA for routine detection of Y‐ST. On the contrary, evaluation of the NAG‐ST ELISA and the SMA using 139 strains of V. cholerae non‐O1 showed discordant results and this was attributed to the presence of the suckling mice active factor(s) such as El Tor hemolysin and to the production of low amounts of NAG‐ST. Concentration of culture supernatants of V. cholerae non‐O1 followed by heating at 100 C was essential to obtain reproducible results by both the NAG‐ST ELISA and the SMA. The ELISA developed in this study can be used for the identification of biologically active strains. While recently genetic methods such as polymerase chain reaction became available and were very reliable and simple techniques, the ELISA in this study has an advantage in detecting biologically toxic gene products of the strains. The genetic methods cannot differentiate silent STa genes which we often encounter in the case of Y. enterocolitica.
Functional Properties of Pro Region of Escherichia coli Heat‐Stable Enterotoxin
Authors: Yamanaka Hiroyasu , Fuke Yasunori , Hitotsubashi Shunji , Fujii Yoshio , Okamoto Keinosuke 
Published: November 2013

AbstractEscherichia coli heat‐stable enterotoxin Ip (STp) is synthesized as the 72‐amino‐acid residue precursor consisting of three regions: pre region (amino acid residues 1 to 19), pro region (amino acid residues 20 to 54), and mature ST (mST) region (amino acid residues 55 to 72). We examined the role of the pro sequence of STp in enterotoxigenicity of a strain by deleting the gene fragment encoding amino acids 22 to 57. This deletion caused a remarkable reduction of its enterotoxic activity of culture supernatant. In order to analyze the sequence responsible for the function of the pro region, two additional deletion mutants were made. The deletion of the sequence covering amino acids 29 to 38, which is conserved in all sequences of ST reported, brought about a significant reduction of enterotoxic activity but the deletion of the non‐conserved sequence (amino acids 40 to 53) did not. This result shows that conserved sequence is mainly responsible for the function. Subsequently, to examine the mechanism of action of the pro region, plasmids carrying DNA sequences of hybrid proteins consisting of pre‐pro‐nuclease, pre‐mST‐nuclease, pre‐pro‐mST‐nuclease and pre‐pro‐nuclease‐mST were constructed. Amino acid sequence determination and SDS‐polyacrylamide gel analysis revealed that these fusion proteins were cleaved between pre sequence and pro sequence during secretion and the cleaved fusion proteins were accumulated in periplasmic space. But the amount of hybrid protein accumulated in the periplasmic space varied among the strains. That is, the amount of the pre‐pro‐nuclease gene product that accumulated in the periplasmic space was the highest of all fusion gene products. These results indicate that the existence of the mST region strongly interferes with the translocation of the gene product into the periplasmic space and that the pro region functions to guide the mST region into the periplasmic space.
Restriction Fragment Length Polymorphism Analysis of Epidemiologically Related Mycobacterium tuberculosis Isolates
Authors: Takahashi Mitsuyoshi , Kazumi Yuko , Fukasawa Yutaka , Hirano Kazue , Mori Toru , Dale Jeremy W. , Abe Chiyoji 
Published: November 2013

AbstractRestriction fragment length polymorphism (RFLP) analysis of a large number of Japanese isolates of Mycobacterium tuberculosis, containing isolates from small outbreaks of M. tuberculosis infection, and clinical isolates of M. bovis BCG, was carried out using a DNA probe derived from the insertion sequence IS986. Clinical isolates of M. tuberculosis had a high degree of RFLP. The occurrences of the IS element varied from 1 to 19, the majority of isolates having 8 to 15 copies. Very similar fingerprints, however, were seen among strains isolated in the Kanto district. In particular, 3 strains were of the same pattern with or without an additional band. Similarity of the banding patterns of strains islated in the same district was observed in other areas. Six groups of strains, each group arising from a suspected common source of infection, were analyzed. Of these, 5 showed identical fingerprints within each group, but one showed different fingerprints. RFLP patterns of three strains isolated from individuals with lymphadenitis developed about two months after BCG vaccination, and one strain isolated from a bladder cancer patient with BCG instillation therapy were identical to those of BCG‐Tokyo which had been used for the vaccination and therapy. These results confirm that RFLP analysis using IS986 is a suitable tool for epidemiology of tuberculosis.
In Vitro Modification of Human Immunodeficiency Virus Type 1 (HIV‐1) Infectivity by the U937 Cells
Authors: El‐Farrash Mohamed A. , Masuda Takao , Kuroda Marcelo J. , Harada Shinji 
Published: November 2013

AbstractThe effect of host cell factors on infectivity of human immunodeficiency virus type 1 (HIV‐1) was studied by infecting a monoblastoid cell line (U937) or a T‐cell line (MOLT‐4) with a highly infective single clone of HIV‐1 and comparing the infectivity of the produced viruses to different cell lines. Chronically infected U937 cells consistently produced viruses with minimal infectivity. This phenotypic change was host‐dependent as the back‐passage of the U937‐produced low infective viruses into MOLT‐4 cells resulted in regaining their original high infectivity. Southern and Northern blot analyses of the HIV‐1 grown in U937 cells did not reveal any genomic difference between it and the virus grown it MOLT‐4 cells. The radioimmunoprecipitation analysis of viral proteins showed that the HIV‐1‐infected U937 cells had a different pattern of envelope glycoproteins and core proteins, which well correlated with the low infectivity of the produced viruses. This experimental system using MOLT‐4 and U937 cell lines would be useful to further explore host cell factor(s) which play an important role in the regulation of HIV‐1 infectivity.