bluuebunny/crossref_metadata_2025 huggingface dataset:
Surface Hydrophobicity of “Rheumatogenic” and “Nephritogenic” Strains of Group A Streptococci and the Ultrastructural Surface Feature of Pharyngeal Cells Exposed to Group A Streptococci
Authors: Kumar Krishnadas Subhadramma Nanda , Ganguly Nirmal Kumar , Anand Inder Singh , Wahi Purshotam Lal
Published: November 2013

The present study was carried out to determine the surface hydrophobicity of group A streptococcal strains responsible for rheumatic fever (RF), “rheumatogenic” strains (RG strains) and strains causing glomerulonephritis, “nephritogenic” strains (NG strains) in relation to their adhesion to human pharyngeal cells. Scanning electronmicroscopic (SEM) studies were carried out to the difference, if any, in the adherence of group A streptococci (M type 5) to pharyngeal and buccal cells (PEC and BEC). By employing two techniques for hydrophobicity determination, salt aggregation titre (SAT) and n‐hexadecane binding‐technique, it was observed that RG strains (M5, Ml and M6) were more hydrophobic than NG strain, M49. However, NG strain M12 was almost equally as hydrophobic as RG strains. The adherence of RG strains, except M1 and M24, to PEC was greater in number than that of NG strains. Although M1 strain was hydrophobic, its adherence to PEC was less. Pepsin and trypsin treatment with streptococci reduced the hydrophobicity and adherence of RG and NG strains to PEC. SEM studies revealed firmly adhered indigenous bacteria on PEC and BEC. Streptococci (M5) adhered more to PEC than to BEC. SEM studies also showed that PEC had a peculiar ultrastructural surface feature to which streptococci adhered. These findings suggest that streptococcal hydrophobicity alone does not determine their adhesion to PEC. The surface nature of PEC might be a characteristic feature of the epithelial cells that allows streptococci to adhere and colonize or it might be a consequence of streptococcal adhesion.
Antibacterial Activity of Cefpodoxime against Branhamella catarrhalis
Authors: Takenouchi Takashi , Nishino Takeshi
Published: November 2013

The antibacterial activity of cefpodoxime against Branhamella catarrhalis was studied. All of the 65 clinical isolates tested were inhibited at and below 1.56 μg/ml, both at 107 and at 105 CFUs. The following was further studied on B. catarrhalis N‐5 which showed average susceptibility to each drug examined. Bactericidal activity was observed at and above the MIC. Scanning and transmission electron microscopy revealed morphological changes, such as cellular swelling, bleb formation, inhibition of septum formation, and lysis, of the cells exposed to cefpodoxime at concentrations around the MIC. Cefpodoxime was poorly hydrolyzed by the β‐lactamase and it showed affinity for two penicillin‐binding proteins that had approximate molecular weights of 83 and 74 kilodaltons, with I50 values of 3.7 and 2.1 μg/ml, respectively.
Characterization of Neurotoxigenic Clostridium butyricum Strain by DNA Hybridization Test and by in vivo and in vitro Germination Tests of Spores
Authors: Takeshi Kouichi , Fujii Nobuhiro , Kimura Kouichi , Yamakawa Kiyotaka , Nakamura Shinichi , Kumagai Mitsuru , Oguma Keiji
Published: November 2013

The germination of spores of a neurotoxigenic Clostridium butyricum strain (BL 6340), which was isolated from infant botulism in Italy, and that of a non‐toxigenic C. butyricum type strain (NCIB 7423) were studied. The spores of BL 6340 strain were killed at 80 C for 10 min, and required the mixture of L‐alanine, L‐lactate, glucose and bicarbonate for their optimal germination. These characteristics are the same as those of Clostridium botulinum type E strain, but different from those of NCIB 7423 strain. In a hybridization test, however, the labeled DNAs extracted from NCIB 7423 strain highly (98%) hybridized to the DNAs of the BL 6340 strain, but little (45%) to the DNAs of C. botulinum type E strain. The biochemical properties of the BL 6340 and NCIB 7423 strains were identical, but different from those of C. botulinum type E. These data confirmed that the BL 6340 strain belongs to C. butyricum species, but that only its characteristics of toxin production, its minimum requirements for germination, and the behavior of its spores to heat treatment are the same as those of C. botulinum type E. When conventionally raised suckling mice were injected with 5 × 107 spores of BL 6340 strain intra‐ or orogastrically, botulism was not observed. However, 8‐ to 13‐day‐old mice had type E botulinum toxin in the large intestine 3 days after introduction of its spores.
Prethymic Nylon Wool‐Passed Bone Marrow Cells, Substituting for Helper T Cells, Can Augment the Generation of Cytotoxic T Lymphocytes from Their Precursors
Authors: Tanabe Masao J.
Published: November 2013

Nylon wool‐passed bone marrow (NW‐BM) cells treated with anti‐Thy.l monoclonal antibody and complement were added to a mixed lymphocyte culture which contained a limiting number of lymph node cells, as responder cells, and a sufficient number of mitomycin‐c‐treated allogeneic spleen cells as stimulator cells. NW‐BM cells of the same MH.C haplotype as responder cells enhanced the generation of allo‐specific cytotoxic T lymphocytes (CTL) not only at a relatively high dose (3 × 103 cells/well) of responder cells, but also at an extremely dilute dose (1 × 103 cells/well). NW‐BM cells which had a third‐party MHC haplotype, a haplotype different from both responder and stimulator cells, also enhanced the generation of CTL at relatively high doses, but not at low doses, of responder cells. NW‐BM cells which had MHC haplotypes identical with those of responder cells induced CTL from helper T cell‐depleted responder cells, but NW‐BM cells which had the third‐party haplotype did not. These results showed that the enhancing effects of NW‐BM cells of the same MHC haplotype as responder cells might be due to a specific helper effect and the enhancing effect of NW‐BM cells of the third‐party haplotype might be due to a nonspecific filler effect, which only conditioned the cultured cells. It was also found that, to exhibit the helper effect, NW‐BM cells had to possess MHC class II, but not MHC class I, molecules in common with CTL precursors. This study showed that in the induction of CTL, prethymic NW‐BM cells had a capability comparable to that of mature helper T cells.
Accumulation of Ascites and Increase in Skin Vascular Permeability Observed by Injection of Adsorbed Diphtheria‐Purified Pertussis‐Tetanus Combined Vaccine in Guinea Pigs
Authors: Goto Norihisa , Kato Hiroshi
Published: November 2013

Intraperitoneal injection of adsorbed diphtheria‐purified pertussis‐tetanus combined vaccine (DPPT) often causes an ascites‐accumulating (A‐A) reaction in guinea pigs. Those vaccines which caused A‐A reaction increase the vascular permeability (VP) of the skin tissue at the intracutaneous injection sites. A total of 23 lots of DPPT were assayed for A‐A and VP‐increasing activities. A positive correlation between the volume of ascites accumulated and the intensity of VP was significant. Other kinds of aluminum‐adsorbed (Al‐Ad) vaccines such as Al‐Ad tetanus and Al‐Ad diphtheria‐tetanus combined and Al‐Ad hepatitis B vaccines induced little or no A‐A reaction. These results were discussed from the viewpoint of quality control in the process of the production of DPPT.
An Indirect Immunofluorescence Method for Detection of Mycoplasma hominis in Vaginal Smears
Authors: Hirai Yoshikazu , Kanatani Tomohisa , Ono Masako , Matsushita Osamu , Kanemasa Yasuhiro
Published: November 2013

We developed a novel method for the detection of Mycoplasma hominis from vaginal swabs using an indirect immunofluorescence technique. It is a rapid and simple method that can be finished in only 5 hr and is more sensitive than the usual culture isolation method. The indirect immunofluorescence method was applied to vaginal smears from 193 healthy women and 33.7% gave a positive test. This value was much higher than that (11.4%) obtained from the same specimens by the culture method. When vaginal smears were subjected to Papanicolaou staining after the indirect immunofluorescence method, the specific immunofluorescence of the epithelial cells was located exactly at the sites of granular aggregates stained with Papanicolaou stain. A histological examination by Papanicolaou staining showed that the incidence of inflammation seems to be slightly higher in M. hominis‐carriers than in non‐carriers.
Synthetic Oligonucleotides with Particular Base Sequences from the cDNA Encoding Proteins of Mycobacterium bovis BCG Induce Interferons and Activate Natural Killer Cells
Authors: Tokunaga Tohru , Yano Osamu , Kuramoto Etsuro , Kimura Yoshimitsu , Yamamoto Toshiko , Kataoka Tetsuro , Yamamoto Saburo
Published: November 2013

Thirteen kinds of 45‐mer single‐stranded oligonucleotide, having sequence randomly selected from the known cDNA encoding BCG proteins, were tested for their capability to augment natural killer (NK) cell activity of mouse spleen cells in vitro. Six out of the 13 oligonucleotides showed the activity, while the others did not. In order to know the minimal and essential sequence(s) responsible for the biological activity, 2 kinds of 30‐mer and 5 kinds of 15‐mer oligonucleotide fragments of an active 45‐mer nucleotide were tested for their activity. One of the 30‐mer oligonucleotides, designated BCG‐A4a, was active, but the other 30‐mer was inactive. All of the 15‐mer oligonucleotide fragments were inactive. The BCG‐A4a also stimulated the spleen cells to produce interferon (IFN)‐α and‐γ An experiment using anti‐IFN antisera showed that the NK cell activation by the oligonucleotide was ascribed to the IFN‐α produced. It was noticed that all of the biologically active oligonucleotides possessed one or more palindrome sequence(s), and the inactive ones did not, with an exception of a 45‐mer inactive oligonucleotide containing overlapping palindrome sequences (GGGCCCGGG). These findings strongly suggest that certain palindrome sequences, like GACGTC, GGCGCC and TGCGCA, are essential for 30‐mer oligonucleotides, like BCG‐A4a, to induce IFNs.
Acceleration of Adenovirus Replication and Increased Virion Production by Treatment with the Steroid Hormone 17 Beta‐Estradiol
Authors: James Calvin B. L. , Vanderpool Eustace A. , Roane Philip
Published: November 2013

We report here that concentration of an estrogen known to promote enhanced transformation and to increase oncogenicity of rat embryo cells, accelerate the production and increase the yield of progeny virions in adenovirus type 12 (Ad 12)‐infected HEp‐2 cells. Further, measurement of the incorporation of radioactive RNA and DNA precursors indicated that macromolecular synthesis in the estrogen‐treated, infected cells was accelerated. Possible explanations for this observation are discussed.
The Role of Pili in Colonization of the Rabbit Intestine by Vibrio parahaemolyticus Na2
Authors: Nakasone Noboru , Iwanaga Masaaki
Published: November 2013

Vibrio parahaemolyticus Na2 and its pili were studied in relation to intestinal colonization. The isolated pili were adhesive to the intestinal epithelium. The adhesion of the organisms was blocked by masking the epithelial receptor with the purified pili, or by treating the organisms with anti‐pilus antibody (Fab fraction). These results suggest that the pili of V. parahaemolyticus Na2 play an important role in the adhesion of the organisms to the rabbit intestine.
Comparison of Indirect Immunofluorescence (IF) Test with Enzyme‐Linked Immunosorbent Assay (ELISA) in Screening of Hybridomas to Very Virulent Marek's Disease Virus
Authors: Nakajima Kazuhiro , Shibayama Takeshi , Naito Matao , Kurimura Takashi , Hirai Kanji
Published: November 2013

For identifying virus‐specific antigens of Marek's disease virus (MDV), monoclonal antibodies (MAbs) against strain Md5 of serotype 1, which is known to be a very virulent MDV (vvMDV), were isolated. Fifty‐eight hybridoma clones that secreted MAbs against vvMDV were obtained. Of these MAbs, 36 gave positive reactions in an immunofluorescence (IF) test, and 22 gave positive reactions on enzyme‐linked immunosorbent assay (ELISA). None of these MAbs gave positive reactions in both the IF test and ELISA. Of the MAbs that gave positive reactions in the IF test, 33 clones reacted with MDV1‐specific epitopes, the other three reacting with MDV1‐HVT intertypic epitopes. None of the clones reacted with MDV1‐MDV2 intertypic epitopes. Three virus‐specific polypeptides were identified by radioimmunoprecipitation and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) or immunoblotting. These polypeptides were recognized by 12 MAbs giving positive reactions by IF, but by none of those giving positive reactions by ELISA. In addition, size heterogeneity of the MDV1‐specific phosphorylated polypeptides in the MDV1 strains was shown using the MAbs against Md5.
Interactions between Immune‐Stimulating Complexes (ISCOMs) and Peritoneal Mononuclear Leucocytes
Authors: Watson Dennis L. , Watson Nikki A. , Fossum Caroline , Lovgren Karin , Morein Bror
Published: November 2013

Studies were undertaken in mice using immune‐stimulating complexes (ISCOMs) or micelles prepared from envelope glycoproteins of human influenza virus (PR8) and matrix (i.e., ISCOM skeleton without incorporated antigen). Electron microscopic studies showed that ISCOMs, in contrast to micelles, have a remarkable affinity for cell membranes and seem to rapidly promote their own internalization by cells to which they adhere. PR8 ISCOMs, but not matrix nor micelles, significantly increased the expression of membrane Ia by peritoneal mononuclear leucocytes 24 hr after intraperitoneal immunization.
Establishment of a Mouse Model of Cystitis and Roles of Type 1 Fimbriated Escherichia coli in Its Pathogenesis
Authors: Liao Jie , Tomochika Ken‐ichi , Watanabe Sadahiro , Kanemasa Yasuhiro
Published: November 2013

The role of type 1 fimbriae in promoting bladder colonization and the course of Escherichia coli cystitis were examined with type 1 fimbriated strains of clinically isolated E. coli. In the experiments of mice in vivo, intact bladder epithelium showed natural resistance to the adherence of type 1 fimbriated and non‐fimbriated E. coli. However, the exfoliation of bladder superficial cells by trypsinization before the bacterial inoculation promoted the adhesion and colonization of type 1 fimbriated E. coli onto bladder epithelium. After colonization of E. coli, maximum numbers of E. coli and leukocytes were observed 3 days after inoculation. Nine days after inoculation, both of E. coli and leukocytes disappeared and the regeneration of superficial cells was observed. On the other hand, superficial cells in mice injected with phosphate‐buffered saline or non‐fimbriated E. coli regenerated 5 days after trypsinization. The present study demonstrated that the removal of superficial cells is essential for the adhesion and colonization of type 1 fimbriated E. coli onto bladder epithelium in vivo and a new model of E. coli cystitis in mice was established. The model which we established is valuable for histopathological, immunological, and therapeutic studies.
Heterogeneity of S‐Layer Proteins of Lactobacillus acidophilus Strains
Authors: Masuda Kuniyoshi
Published: November 2013

An S‐layer (surface regular array) was found in the cell wall from six out of ten strains of Lactobacillus acidophilus examined by electron microscopic observations. All of the six strains which were shown to carry the S‐layers belonged to the deoxyribonucleic acid (DNA) homology group A, but not to B, which had been classified by Johnson et al (Int. J. Syst. Bacteriol. 30: 53–68, 1980). On the other hand, the other four strains which possessed no S‐layers were in the homology group B. The apparent molecular weights of the S‐layer proteins ranged from 41 to 49 kDa as estimated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Of the S‐layer proteins from the six strains, three were susceptible to chemical cleavage with N‐chlorosuccinimide, giving different peptide maps. All of the six S‐layer proteins were fragmented by limited proteolysis with Staphylococcus aureus V8 protease, and gave markedly different peptide patterns by the subsequent peptide mapping analysis, except that the peptide maps of the S‐layer proteins from the two strains which were in the same subgroup were identical.
Role of Type 1 and Type 3 Fimbriae on the Adherence and Pathogenesis of Salmonella enteritidis in Mice
Authors: Aslanzadeh Jaber , Paulissen Leo J.
Published: November 2013

Through hemagglutination tests two isogenic strains of Salmonella enteritidis were shown to possess type 1 fimbriae (strain V) and type 1 and type 3 fimbriae (strain A). The two strains bound to human buccal and mouse small intestine epithelial cells. Strain A attached to the epithelial cells more readily and in larger numbers in comparison to strain V. Adherence of both strains were sensitive to the presence of D‐mannose and pretreatment of the epithelial cells with tannic acid did not promote D‐mannose resistant type binding of strain A S. enteritidis to human buccal and mouse small intestine epithelial cells. Furthermore, results from LD50 study indicated that, when the tests were carried out through oral inoculation of the mice the highly fimbriated stain A appeared to be more virulent. However, when the tests were carried out through intraperitoneal inoculation strain V was more virulent. These results indicate that adherence is a major contributing factor to the virulence of S. enteritidis and both type 1 and type 3 fimbriae contribute to this phenomenon.
Detection of Leishmania Antigen in Kala Azar Patients Using Monoclonal Antibodies
Authors: Sinha Raghu , Arora Sunil K. , Datta Usha , Sehgal Shobha
Published: November 2013

Twenty‐one monoclonal antibodies were produced against promastigote antigens of Leishmania donovani, Five monoclonal antibodies (Hyb.17, 6, 5, 4 and 2) identifying molecules associated with various L. donovani antigenic determinants ranging from 42–116 kDa were selected as ‘capture antibodies’ and compared with specific anti‐leishmania antisera for detection of circulating leishmania antigens in kala azar patients' sera in a competitive‐enzyme‐linked immunosorbent assay system (ELISA). The anti‐leishmania antisera could detect circulating antigen in 30% of kala azar cases while out of the five monoclonals, Hyb.17 could effectively detect circulating leishmania antigen in 85.4%. The efficacy of Hyb.6 was however low (31.7%). The antigens recognized by these monoclonal antibodies in the western blot assay could possibly represent the ones circulating in sera of patients suffering from kala azar. A cocktail of these monoclonal antibodies may be more useful than the conventional polyclonal antisera in detection of circulating antigen for clinical diagnosis of kala azar.
Recognition of the Self Idiotype by T Cells: Induction of a Rapid Increase in Cytoplasmic Free Calcium in T Cells Recognizing a Variable L Chain Determinant
Authors: Utsunomiya‐Tate Naoko , Nakanishi Mamoru , Arata Yoji , Sugiyama Haruo , Vera‐Antola Maria E. , Fujio Hajime , Sakato Nobuo
Published: November 2013

To investigate the initial stages of recognition of the self idiotype (Id) by T cells, we examined the early increase in cytoplasmic free calcium ([Ca2+]i) occurring in murine CD4+ T cells specific for a model Id, Id315, following their interaction with the Id. The changes in [Ca2+]i were monitored with stopped‐flow fluorometry bv loading T cells with fura 2, a Ca2+‐binding fluorescent dye. An increase of [Ca2+]i in the Id‐specific T cell line was dependent on the presence of both antigen‐presenting cells (APC) and Id315. When T cells were mixed with APC pulsed with M315 for 90 min at 37 C, a significant increase in T cell [Ca2+]i was observed within one second. A pronounced elevation in [Ca2+]i was also observed in T cells after their interaction with APC which had been pulsed for 90 min with VL‐315 Id‐containing proteins (such as VL‐315, L315, FV‐315 or Fab′‐315 fragments). In contrast, pulsing APC for 5 min with the VL fragment produced little or no change in the [Ca2+]i. These results suggest that VL must be further processed by APC before it can be recognized by T cells. Indeed, a synthetic VL region peptide (positions 91–108, designated as P18) produced an elevation in T cell [Ca2+]i when mixed with APC without pulsing.
A Simple Purification Method of Vibrio cholerae Non‐O1 Hemagglutinin/Protease by Immunoaffinity Column Chromatography Using a Monoclonal Antibody
Authors: Naka Atsuko , Honda Takeshi , Miwatani Toshio
Published: November 2013

A new simple purification method (I) for Vibrio cholerae non‐O1 hemagglutinin/protease (NAG‐HA/P) was developed. The method (I) requires only an immunoaffinity column chromatography using a monoclonal antibody against NAG‐HA/P. The method (I) is much simpler than previously reported purification method (II) (Honda, T. et al, Infection and Immunity 57: 2799–2803, 1989) which required four or more complicated chromatographic procedures. Method (I) also gave an improved recovery rate (about 27%) compared with (II), The molecular weight of NAG‐HA/P purified by method (I) was mainly 34 kilodaltons (kDa) with a little of 32 kDa, whereas that of NAG‐HA/P purified by (II) was usually 32 kDa. Immunological analysis by the Ouchterlony double gel diffusion test and Western blotting test using polyclonal antibody against 32 kDa protein revealed that the 34 and 32 kDa proteins are immunologically indistinguishable and thus it is supposed that 34 K protein is an isoform or a preform of the 32 K protein.
Relationship between an 85 kDa Protein and the Protective Effects of Mycoplasma pneumoniae
Authors: Yayoshi Masami , Sasaki Tsuguo , Yoshioka Morimasa
Published: November 2013

In the immunoblot analysis, sera from patients infected with Mycoplasma pneumoniae reacted with the 168 kDa (P1) and the 85 kDa proteins of virulent strain FH‐P24 and P24‐S1 mutant strain but not with the 85 kDa protein of P24‐S11. Sera of hamsters and BALB/c mice, which had been immunized with live vaccines, were tested. In FH‐P24 immunized animals, 100% or 80%, and in P24‐S1, 40% of hamsters and 60% of BALB/c mice, developed antibodies against the 85 kDa protein, but antibodies were not detected in sera of P24‐S11 immunized animals. The correlation between the development of antibodies to 85 kDa protein in the sera of vaccinated animals and the effects of protection by living vaccines were suggested.
Genetic Variation among Malaysian Isolates of Salmonella typhi as Detected by Ribosomal RNA Gene Restriction Patterns
Authors: Pang Tikki , Altwegg Martin , Martinetti Gladys , Koh Chong Lek , Puthucheary Savithri
Published: November 2013

Genetic variation among Malaysian isolates of Salmonella typhi was determined by analysis of ribosomal RNA gene restriction patterns. Of the 20 isolates analyzed, eight different pattern combinations were detected. The amount of variation observed was also dependent upon the restriction endonuclease used; PstI produced more different patterns than did SmaI. The results suggested that disease activity was due to a number of different clones circulating simultaneously rather than a single strain. Further implications of the data are discussed.
Sequence Comparison of the Capsid Region of Hepatitis E Viruses Isolated from Myanmar and China
Authors: Aye Thein Thein , Uchida Toshikazu , Ma Xuezhung , Iida Fusae , Shikata Toshio , Zhuang Hui , Win Khin Maung
Published: November 2013

Hepatitis E viruses (HEVs) were isolated during epidemics, one from Myanmar (formerly called Burma) and one from China and were partially sequenced. Another HEV Myanmar strain from sporadic hepatitis was previously sequenced by us. A cDNA sequence comparison was performed among them in the 3′‐terminal region, approximately 750‐base long. This region contained at least two immunological epitopes and was considered to correspond to the structural protein. The nucleotide sequence identity was 97.2% between the two Myanmar strains and 93.3 and 92.5% between the two Myanmar and the China strain. The deduced amino acid sequence identity ranged from 98.4 to 100.0% among the three strains. Thus this segment was well conserved on the amino acid level among the different strains isolated from these two Asian countries, although the China strain diverged more from the Myanmar strains on the nucleotide sequence level. This data may provide important information for the development of a vaccine and for identification of the virological link between different geographical locations.
Heat‐Labile Toxin from Bordetella parapertussis Induces Contraction of Smooth Muscle Cells in Culture
Authors: Nagai Masaaki , Endoh Masahiko , Burns Drusilla L. , Nakase Yasukiyo
Published: November 2013

The ability of Bordetella heat‐labile toxin (HLT) to contract various types of cells in culture was examined. HLT from B. parapertussis induced contraction of cultured smooth muscle cells from trachea, intestine, uterus and vas deferens as well as from aorta. The time required for contraction decreased as the dose of B. parapertussis HLT increased from 3 to 100 MID/ml. Upon exposure of cells to concentrations of toxin greater than 100 MID/ml, at least 2 hr was required for contraction. HLT from B. parapertussis did not affect cultured cardiac or skeletal muscle cells within 8 hr after the exposure to HLT (100 MID/ml). No effect on other types of primary culture cells or established cells such as Chinese hamster ovary (CHO) cells has been described. These data indicate that the primary target cells for HLT might be smooth muscle cells.
Characterization of an Exo‐β‐d‐Fructosidase from Streptococcus mutans Ingbritt
Authors: Igarashi Takeshi , Yamamoto Ayako , Goto Nobuichi
Published: November 2013

An extracellular enzyme β‐d‐fructosidase was purified from the culture supernatant of Streptococcus mutans Ingbritt and characterized. The molecular weight of the enzyme was 127,000 as determined by SDS‐polyacrylamide gel electrophoresis. The enzyme was specific for levan which mainly consists of β‐(2,6)‐linked d‐fructose and was also able to hydrolyze inulin, sucrose and raffinose at the activities of 13, 9 and 5% of that hydrolyzing levan, respectively. The pH optima for levan, inulin and sucrose were approximately 5.5, 6.0 and 5.0, respectively. The enzyme was optimally reactive at 55 C for levan. The enzyme was inhibited by Fe3+, Hg2+ and Zn2+ and not by either anionic or non‐ionic detergents. Paper chromatographic analysis revealed that the enzyme attacked levan by an exo‐type mechanism.
Structural Similarity of the HLA‐DQ Region in DQ3 and DQ4 Haplotypes and Structural Diversity of the HLA‐DQ Region in HLA‐DR7 Haplotypes
Authors: Tate Genshu , Ishizawa Mitsugu
Published: November 2013

Genomic DNA obtained from a B lymphoblastoid cell line was digested with appropriate restriction endonuclease and hybridized with several probes specific for genes encoding HLA‐DQ. Southern hybridization with a DQA1 3′untranslated (UT) region probe showed DQ2‐type hybridization pattern in DR7DQ3 haplotype. On the contrary, DQB1 3′UT probe showed DQ3‐type pattern in the same haplotype. Gene cloning and DNA sequencing analysis revealed a repetitive sequence, (TG)19, between DQA1 and DQB1 gene in the DR7DQ3 haplotype. These results suggest that a recombination event has occurred near this potential Z‐DNA structure in the haplotype, DR7DQ3. The 3′UT region probes of DQA1 and DQB1 genes failed to detect restriction fragment length polymorphism (RFLP) differences between DR4DQ3 and DR4DQ4 haplotypes in this experiment, suggesting that the gene structure between DQA1 and DQB1 is conserved in these haplotypes.
Comparison of Immunological Effects of Cholera Toxin on Autoimmune MRL/lpr and BXSB Mice
Authors: Zhou Nan‐nan , Nakai Shoji , Oka Mikio , Nagasawa Hideyuki , Himeno Kunisuke
Published: November 2013

MRL/lpr and BXSB mice were treated weekly or biweekly with cholera toxin (CT) in intravenous dose of 2 μg/mouse. CT treatment notably alleviated proteinuria in MRL/lpr mice, but did not influence the course of lupus nephritis in BXSB male mice. Flow cytometric analysis showed that anomalous B220+ T cells in spleen and thymus were reduced in CT‐treated MRL/lpr mice while no significant change in lymphocyte populations was induced in BXSB male mice by this treatment. The suppressive effect of CT treatment on Con A response and the augmentative action on LPS response were observed in MRL/lpr mice. The latter may reflect increased B cells in relative number in the peripheral lymphoid organs. Mitogenic responses in CT‐treated BXSB male mice remained unchanged in comparison with those of untreated group. Increased production of IL‐6 by spleen cells was demonstrated in MRL/lpr mice treated with CT while in BXSB mice the level of IL‐6 was not changed by the treatment with CT. Production of IFNγ was suppressed by CT treatment in both strains of mice. This may be attributed to the inhibitory effect of CT on IFNγ‐producing Th1 cells as reported previously (Munoz et al, J. Exp. Med. 172: 95–103, 1990). However, CT treatment did not inhibit anti‐DNA antibody production in BXSB mice, whereas the autoantibodies were markedly decreased in MRL/lpr mice treated with CT.
Extracellular Localization of a Nonfimbrial Hemagglutinin of Yersinia pseudotuberculosis
Authors: Takumi Kenji , Koga Tetsuro , Oka Tatsuzo , Tsuji Hideaki , Tsubokura Misao , Sheng Yao Jian
Published: November 2013

A nonfimbrial hemagglutinin (HA) of Yersinia pseudotuberculosis was observed by immunoelectron microscopy using monospecific HA antiserum and protein A‐gold conjugate. The HA, an amorphous but morphologically identifiable entity, was located in a region distal to or detached from the outer edge of bacteria.
Production of a Monoclonal Antibody That Recognizes the Lipopolysaccharide of a Campylobacter‐Like Organism
Authors: Kuan Steven Kok Chee , Coloe Peter John , Alderton Malcolm Raymond
Published: November 2013

A monoclonal antibody was produced to a Campylobacter‐like organism (RMIT 32A) which was isolated from the terminal ileum of a pig with proliferative enteritis. Isotyping of the antibody revealed that it was an IgG2a with kappa light chains. Immunoblots using the antibody against proteinase‐K‐treated whole cell lysates of RMIT 32A, a selection of Campylobacter species and other enteric bacteria showed that the antibody was specific for RMIT 32A and was directed against the lipopolysaccharide. This antibody can be used for the specific detection of RMIT 32A.
Spontaneous Proliferation of HTLV‐II‐Infected Peripheral Blood Lymphocytes: HLA‐DR‐Driven, IL‐2‐Dependent Response
Authors: Lal Renu B. , Hjelle Brian , Rudolph Donna L.
Published: November 2013

Peripheral blood lymphocytes obtained from HTLV‐II‐infected persons (n = 13) and cultured in the absence of exogenous stimulator demonstrated augmented spontaneous proliferation (17,672 ± 5,498 cpm) when compared with cells from healthy donors (1,921 ± 1,306 cpm). Removal of non‐T population did not abrogate the proliferative response of patients' PBMC, suggesting that the proliferation is not related to the autologous mixed lymphocyte reaction. Addition of recombinant interleukin‐2 (rIL‐2; 0.1 U/ml) to spontaneously proliferating cultures from HTLV‐II‐infected persons resulted in a 3‐ to 4‐fold increase in proliferation (61,985 ± 16,003); in contrast, PBMC from controls demonstrated 38‐ to 42‐fold increase in their proliferative capacity in response to rIL‐2 (77,256 ± 13,044). Antibodies to both IL‐2 receptor and HLA‐DR were able to inhibit the spontaneous proliferation of PBMC from HTLV‐II‐infected persons in a dose‐dependent manner. Furthermore, addition of cyclosporin A, which preferentially blocks accumulation of IL‐2 mRNA, also inhibited spontaneous proliferation in a dose‐dependent manner. These observations suggest that the spontaneous proliferation of HTLV‐II‐infected PBMC is at least in part an HLA‐DR‐driven, IL‐2‐dependent event, which is not analogous to the AMLR.
Influence of Inoculation Route on Virulence of Rhodococcus equi in Mice
Authors: Takai Shinji , Sasaki Yukako , Tsubaki Shiro
Published: November 2013

Viruleuce of R. equi ATCC 33701 was compared by the intraperitoneal (i.p.) and intravenous (i.v.) routes in mice. Strain ATCC 33701 was more virulent by the i.v. than the i.p. route. The LD50 of strain ATCC 33701 by either route correlated with the initial number of bacteria in the liver and spleen at day 0.
Correlative Detection of Human Immunodeficiency Virus (HIV) Antigen p24 and Epstein‐Barr Virus DNA In Vitro: Clinical Influence on HIV Infection
Authors: Lardelli Pilar , Manzano Diego , Steinberg Seth M. , Madariaga Lucila , Antón Isabel , Cisterna Ramón
Published: November 2013

The existence of molecular transactivations between EBV and HIV‐1, as well as reactivations of EBV latent infections in AIDS patients, have been recently documented. In order to shed more light on the putative association between EBV and HIV, and its role in the evolution to AIDS, we have determined simultaneously p24 protein and EBV DNA in culture supernatants of peripheral blood mononuclear cells from 47 individuals suspected of having HIV infection. The results of the in vitro assays were correlated with the clinical stage of the individuals and their serologic status to EBV. Statistical analysis showed a concordance between HIV infection and in vitro detection of EBV DNA (P < 0.002); particularly, a strong correlation between the presence of EBV DNA and p24 in culture was observed (P < 0.001). These results are consistent with the occurrence of viral interactions, manifested in vitro. However, in our series, the appearance of EBV DNA in culture was not concomitant with an elevation of anti‐VCA IgG titers, anti‐EA titers or the development of symptomatology, suggestive of a reactivation of a latent EBV infection or a progression of HIV infection. Therefore we conclude that, although interaction between both viruses may take place at the molecular level, there is no clear evidence of the repercussion that this event may have on the clinical course of HIV infection.
Structural Studies of Peptidoglycans in Campylobacter Species
Authors: Amano Ken‐ichi , Shibata Yoshiko
Published: November 2013

Peptidoglycans (PG) from Campylobacter coli, Campylobacter jejuni, and Campylobacter fetus were composed of muramic acid, glucosamine, alanine, glutamic acid, and diaminopimelic acid in a molar ratio of 1.1: 1: 1.7: 1.1:0.9. Thirty percent of the amino groups of diaminopimelic acid were involved in cross‐linkages between peptides. During cultivation, C. coli and C. jejuni changed from a spiral to a coccoid form. In C. coli, we could isolate PG only from the spiral forms in yields of 0.8‐1.2% by dry weight. C. fetus did not change to a coccoid form, and always contained PG. Thus, it is possible that the morphological transformation from the spirals to the coccoid forms of C. coli and C. jejuni is accompanied by, and probably due to, the degradation of PG.
Naturally Occurring SLE Anti‐DNA Antibodies Recognize Unique Conformation on DNA‐Lysine Photoadduct
Authors: Alam Khurshid , Islam Najmul , Hasan Rashid , Ali Asif , Ali Rashid
Published: November 2013

Native calf thymus DNA has been covalently modified with lysine under UV‐A light. Human autoantibodies on purification through affinity column of native DNA linked to polylysyl‐Sepharose 4B showed almost equal recognition of DNA and photoadduct. The recognition of DNA‐lysine photoadduct by the affinity‐purified autoantibodies might be helpful in understanding their origin in SLE vis‐à‐vis the role of positively charged amino acids in the pathogenesis of autoimmune diseases.
The Effects of Magnesium, Calcium, EDTA, and pH on the In Vitro Adhesion of Staphylococcus epidermidis to Plastic
Authors: Dunne Wm. Michael , Burd Eileen M.
Published: November 2013

The effects of increasing concentrations of magnesium (Mg2+), calcium (Ca2+) or EDTA, and pH on the adhesion of five slime‐positive strains of Staphylococcus epidermidis (Se+) to plastic were examined using an in vitro microwell assay. The addition of Mg2+ (as either MgSO4 or MgCl2) to the bacterial suspension in concentrations as low as 16 μM significantly enhanced the adhesion of all test strains to plastic (P<0.001). Similarly, the addition of Ca2+ (as CaCl2) in concentrations exceeding 128 μM produced a significant increase in the adhesion of all test strains, but not to the extent observed with Mg2+. In contrast, the adhesion of all test strains to plastic was significantly reduced in the presence of EDTA at concentrations greater than 8 mM. However, EDTA in concentrations as low as 0.25 mM caused a significant decrease in the adhesion of two strains of Se+. The effect of pH was variable, but at a pH of 5.0 and 6.0, the adhesion of all test strains was significantly reduced compared to control values at a pH of 7.0. Two strains showed a significant increase in adhesion at a pH of 8.0. We also compared the effects of these variables on the adherence of a slime‐negative phase variant derived from a slime‐positive parent strain. With the exception of pH, the adhesion of both strains in response to increasing divalent cations or EDTA was similar. These data indicate that, in addition to hydrophobic interactions, ligand‐specific binding, and slime production, pH and divalent cations, especially Mg2+, are important determinants of the adhesion of S. epidermidis to plastic surfaces in vitro. This has possible implications in the pathogenesis of biomedical implant infections caused by these organisms.
Co‐Existence of Colonies with Different Serotypes and Other Biological Characteristics in Clinical Isolates of Pseudomonas aeruginosa
Authors: Kobayashi Intetsu , Hasegawa Miyuki , Nishida Minoru , Goto Sachiko
Published: November 2013

The biological characteristics of individual colonies of Pseudomonas aeruginosa from 138 specimens were investigated. Of these isolates, 90 (65.2%) formed colonies of similar appearance and morphology, and 48 (34.8%) formed colonies which differed either in appearance or morphology. The individual colonies of 138 isolates were tested for serotype. The former 90 isolates formed only the colonies with one kind of serotype, whereas 17 of the latter 48 isolates formed the colonies with more than one kind of serotype. All the 9 isolates tested also differed in other biochemical characteristics: acid productions from xylose, mannitol and maltose, urease production and gelatin liquefaction. β‐Lactamase activity was investigated in 7 isolates forming colonies with more than one serotype. There were no marked differences in β‐lactamase activity among the different colonies in 5 isolates but marked differences among those in the other 2 isolates.
Melibiose Transport System in Lactobacillus plantarum
Authors: Tamura Chiyuki , Matsushita Osamu
Published: November 2013

Lactobacillus plantarum ATCC 8014 grew on melibiose at 30 C, but not at 37 C, although it grew on galactose or lactose at either temperature. ATCC 8014 grown on lactose at 30 or 37 C accumulated melibiose slowly, suggesting that melibiose may partly be transported by a lactose transport system. A lactose‐ negative mutant, NTG 21, derived from ATCC 8014 was isolated. The mutant was totally deficient in lactose transport, but retained normal melibiose transport activity. In NTG 21, the melibiose transport activity was induced by melibiose at 30 C, but not at 37 C. The transport activity itself was found to be stable for at least 3 hr at 37 C, suggesting that the induction process in the cytoplasm rather than the inducer entrance is temperature‐sensitive in the organism. The organism also failed to form α‐galactosidase at 37 C when grown on melibiose. The enzyme synthesis, however, was induced by galactose in NTG 21 (and also by lactose in ATCC 8014) even at 37 C, indicating that the induction of the enzyme is essentially not temperature‐sensitive. In NTG 21, melibiose transport system and a‐galactosidase were induced by galactose, melibiose and o‐nitrophenyl‐α‐d‐galactopyranoside when the strain was grown at 30 C. Raffinose induced melibiose transport system only a little, while it was a good inducer for α‐galactosidase. Inhibition studies revealed that galactose may be a weak substrate of the melibiose transport system; no inhibition was demonstrated with lactose and raffinose.
Killing of Escherichia coli by Mononuclear Phagocytes and Neutrophils Stimulated In Vitro with β‐1,3‐d‐Polyglucose Derivatives
Authors: Busund Lill‐Tove Rasmussen , Pål Øian Zbigniew Konopski , Seljelid Rolf
Published: November 2013

Human monocytes, human peritoneal macrophages, mouse peritoneal macrophages and human peripheral neutrophils pretreated with β‐1,3‐d‐polyglucose derivatives showed pronounced bactericidal capacity to Escherichia coli compared to control cells. The increased bactericidal capacity was detectable in mononuclear phagocytes over a wide range of concentrations of bacteria. Granulocytes, however, showed bactericidal capacity only at low concentrations of bacteria. The pretreated mononuclear phagocytes released significant amounts of IL‐1 and PGE2. However, there was no significant release of tumor necrosis factor (TNF). By incubating unstimulated cells with purified IL‐1 and TNF, the bactericidal activity of neutrophils and mononuclear phagocytes was enhanced. Our data indicate that the inability of neutrophils stimulated with β‐1,3‐d‐polyglucose derivatives to kill large numbers of bacteria could be overcome by a combined treatment with purified IL‐1 or TNF in addition to β‐1,3‐D‐polyglucose derivatives. By incubating unstimulated cells with medium from β‐1,3‐d‐polyglucose‐treated human peritoneal macrophages, the bactericidal activity of the cells was enhanced to the same extent as cells pretreated with purified TNF and IL‐1. Cells incubated with IL‐1‐depleted medium from β‐1,3‐d‐polyglucose‐treated human peritoneal macrophages, showed reduced bactericidal activity compared to cells incubated with undepleted medium. These studies demonstrate that β‐1,3‐d‐polyglucose‐treated mononuclear phagocytes and neutrophils show enhanced bactericidal activity. The enhanced activity is partly caused by stimulation of the cells with IL‐1 released from mononuclear phagocytes and partly by other unknown effects of β‐1,3‐d‐polyglucose derivatives on both mononuclear phagocytes and neutrophils.
Changes with Time in the Oral Microflora and Dental Caries Induction in Hyposalivated Rats Fed on Sucrose Diet
Authors: Ooshima Takashi , Yoshida Toshihiko , Aono Wataru , Takei Tsutomu , Izumitani Akira , Sobue Shizuo , Hamada Shigeyuki
Published: November 2013

The effects of hyposalivation on the induction of dental caries and the change in oral microflora were examined at weekly intervals in Sprague‐Dawley rats fed on diet 2000 containing 56% sucrose. In hyposalivated rats, significant dental caries was induced within one week and its severity increased with the experimental period. Bacteriological examinations demonstrated that the number of total cultivable microorganisms, lactobacilli and Staphylococcus aureus increased significantly shortly after surgical induction of hyposalivation, while the number of streptococci and yeasts did not increase significantly until the 7th week, beyond which time remarkable gross caries developed. A positive correlation was found between the caries score and the recovery of lactobacilli from mandibles of hyposalivated rats, while there was no statistically significant correlation between the caries score and the recoveries of S. aureus. On the other hand, dental caries was not induced in control rats fed on sucrose diet with no surgically‐induced hyposalivation. It was also found that the number of lactobacilli increased significantly shortly after diet 2000 was given to control rats, but S. aureus was rarely recovered from the mandibles of control rats throughout the experiments. The roles of lactobacilli and S. aureus in the induction of dental caries under the hyposalivated condition were discussed and it was suggested that lactobacilli may play some significant role in the induction of dental caries in hyposalivated rats.
Identification of Oklahoma Isolate as a Strain of Pseudomonas pseudomallei
Authors: Yabuuchi Eiko , Kosako Yoshimasa , Arakawa Michio , Hotta Hisako , Yano Ikuya
Published: November 2013

Based on the morphological, physiological, biochemical, and nutritional characteristics, cellular fatty acid and lipid composition, ubiquinone‐8 as the major respiratory quinone, guanine‐plus‐cytosine content of DNA, DNA‐DNA homology value, and sequence alignment of 16S rRNA nucleotides, Oklahoma isolate was reidentified as a strain of Pseudomonas pseudomallei.
Neutrophil Response to Pseudomonas aeruginosa in Respiratory Infection
Authors: Ahmed Kamruddin , Dai Tran Cong , Ichinose Akitoyo , Masaki Hironori , Nagatake Tsuyoshi , Matsumoto Keizo
Published: November 2013

Sputum from patients with acute exacerbation of respiratory infection by Pseudomonas aeruginosa was observed under the electron microscope. External to the cell wall of P. aeruginosa a granular, electron‐dense material was observed which is suggestive of capsule. It is supposed that stabilization of capsule occurred by the host antibody, which was produced due to chronic infection by P. aeruginosa. Mucoid type of microcolonies were observed with a fibrous matrix of exopolysaccharide. Other types of microcolonies were surrounded by granular substances or fine fibers. Neutrophil was found to be partially surrounding the microcolony in an attempt to defense. Debris was formed mainly by the destruction of the neutrophil. Most neutrophils were found full of phagocytosed debris; in contrast only a few neutrophils were found to have phagocytosed P. aeruginosa. This study concludes that instead of phagocytosing bacteria, neutrophil phagocytosed debris and bacteria were not completely eradicated. Therefore, this might be one of the factors in the pathogenesis of respiratory infection and persistent colonization by P. aeruginosa.
Comparison of Virulence between Seoul Virus Strain SR‐11 and Hantaan Virus Strain 76–118 of Hantaviruses in Newborn Mice
Authors: Yoo Yung Choon , Yoshimatsu Kumiko , Yoshida Ryu , Tamura Mizuho , Azuma Ichiro , Arikawa Jiro
Published: November 2013

Virulence of hantavirus strain of SR‐11 Seoul virus and Hantaan 76–118 (HTN) of Hantaan virus were compared. Infections of both strains were lethal in newborn mice. However, inoculum required to cause lethal infection was about 4,000 times higher for strain HTN (1.65 × 103 PFU/mouse/LD50) than for strain SR‐11 (0.36 PFU). Thus, both strains were considered pathogenic to newborn mice but they possessed different levels of virulence. The assay system used for these strains in newborn mice proved to be useful in the study of hantavirus vilurence. Growth curves of the two strains in CV‐7 cell cultures were compared. Strain SR‐11 was shown to have higher activity of virus replication and virus release into the culture fluids than strain HTN. The possibility of a relationship between replication activity and high levels of virulence in mice was suggested.