bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence of the Antibody to Influenza C Virus in a Northern Luzon Highland Village, Philippines
Authors: Nishimura Hidekazu , Sugawara Kanetsu , Kitame Fumio , Nakamura Kiyoto , Sasaki Hideo
Published: November 2013

A total of 101 serum samples were collected from the persons (1 to 85 years of age) living in a Philippine mountain village where the contact with other communities has largely been restricted. These sera were tested for the presence of antibody to influenza C virus with hemagglutination‐inhibition and radioimmunoprecipitation tests. The results showed that all the subjects, including the persons who had never been outside the village, contained the antibody to the surface glycoprotein of the virus, and that the age of acquisition of the antibody was significantly lower in this village than in any of the previously studied communities. Thus it appeared that infection with influenza C virus was prevalent even in this small mountain village, presumably with a higher incidence than in the larger, industrialized communities.
Synthesis of Sterols and 5‐Lipoxygenase Products are Required for the G1‐S Phase Transition of Interleukin‐2‐Dependent Lymphocyte Proliferation
Authors: Hata Shingo , Sugama Kazushige , You‐Li Zu , Hatanaka Masakazu , Namba Yuziro , Hanaoka Masao
Published: November 2013

A murine killer T cell line, G‐CTLL 1, whose proliferation depends on the presence of interleukin 2 (IL‐2), was used to analyze the mechanism of IL‐2 action with respect to sterol synthesis and arachidonate metabolism. De novo sterol synthesis was substantially enhanced much earlier than DNA synthesis, and the rate reached a maximum at 13 hr after the addition of IL‐2. Compactin, which is a potent competitive inhibitor of 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase (HMG CoA reductase, the enzyme in the rate‐limiting step of the sterol synthesis), inhibited the IL‐2‐induced DNA synthesis. The addition of mevalonate, the product of HMG CoA reductase, prevented the inhibition of DNA synthesis by compactin, suggesting that the supply of a sufficient amount of sterol is an essential prerequisite for IL‐2 action. The IL‐2‐induced DNA synthesis was also inhibited by AA861, a specific inhibitor of arachidonate 5‐lipoxygenase, and by other lipoxygenase inhibitors such as nordihydroguaiaretic acid and esculetin. In contrast, indomethacin, an inhibitor of arachidonate cyclooxygenase, had no effect. These findings suggest that synthesis of 5‐lipoxygenase products is also a prerequisite. The inhibition of DNA synthesis was effectively inhibited only when compactin or lipoxygenase inhibitors were added early enough to block the synthesis of sterols or 5‐lipoxygenase products; addition of the reagents after 3 hr decreased the inhibition with time. Therefore, about 3 hr after the addition of IL‐2, several drastic intracellular changes are assumed to begin and to lead to DNA synthesis.
Hemagglutination by Pilus Antigen 987P of Enterotoxigenic Escherichia coli
Authors: Ike Kazunori , Nakazawa Muneo , Tsuchimoto Mayumi , Ide Seiya , Kashiwazaki Mamoru
Published: November 2013

The hemagglutination (HA) by pilus antigen 987P of an enterotoxigenic Escherichia coli strain 987 was examined using fresh and glutaraldehyde (GA)‐fixed erythrocytes (RBC) of various animals. Only when GA‐fixed RBC was employed. a strain 987 exhibited striking HA activities. This was also demonstrated by using latex heads sensitized with the 987P antigen. The 987P‐specific antiserum inhibited HA of strain 987 and 987P sensitized latex beads against GA‐fixed RBC. We concluded that HA of strain 987 against GA‐fixed RBC was specifically associated with the presence of 987P pilus antigen but do not exclude a possibility that adhesin is distinct from pili antigen.
Electron Microscopic Demonstration and Isolation of Ribosomes in Mesosomes from Staphylococcus aureus
Authors: Nakasone Noboru , Kawata Tomio , Masuda Kuniyoshi
Published: November 2013

Mesosomes of Staphylococcus aureus 209P were observed to be extruded as tubules upon protoplast formation by electron microscopy and isolated under hypertonic conditions to maintain their structural integrity by differential centrifugation followed by sucrose density gradient centrifugation. Isolated mesosomes were composed of long, branched tubules of irregular sizes and they were shortened during purification. Thin sections of isolated mesosomes showed that the mesosomal tubule was surrounded by a triple‐layered membrane and contained ribosome‐like particles in diameter of about 15 to 20 nm. These particles were isolated from purified mesosomal preparation by disrupting the mesosomal tubule with deoxycholate and Triton X‐100 under hypotonic conditions followed by a linear sucrose density gradient centrifugation. Negatively stained preparations of the isolated particles revealed the same appearance as those of the ribosomes isolated from the cytoplasm. The mesosomal particles sedimented at 70S in sucrose gradients in the presence of 10 mm Mg2+, but they were dissociated into two subparticles, 50S and 30S subunits, upon lowering the Mg2+ concentration to 1 mm. These findings indicate that the mesosomal tubule is packed with ribosomes.
Total Nucleotide Sequences of the Infectious Cloned DNAs of Bean Golden Mosaic Virus
Authors: Morinaga Tsuto , Ikegami Masato , Shimotohno Kunitada , Miura Kin‐Ichiro
Published: November 2013

Complete nucleotide sequences of the infectious cloned DNA components (DNA A and B) of bean bolden mosaic virus were determined. The DNA A (2585 nucleotides) and DNA B (2647 nucleotides) have little sequence homology with each other, but both A and B contain a common region of 205 nucleotides. A possible large hairpin structure is detected in the common region. Nucleotide sequences of DNAs A and B revealed the presence of 8 potential coding regions for proteins (m.w.> 10,000). Among them, four open reading frames (ORFs 1–4) encode proteins of m.w. 30,000 or greater, and are individually coded in virion DNA A and B senses (+) and their complementary senses (‐), respectively. The other four ORFs 5–8 are in virion DNA B(+) and its complementary sense (‐). All of the ORFs 1–4 have regulatory signals for RNA synthesis (TATAA/T) in the region 5′ upstream from a potential start codon ATG.
Differential Recognition of Oral Indigenous Bacteria by Salivary Immunoglobulins A and G
Authors: Deslauriers Noëlla , Seguin Jacynthe , Trudel Luc
Published: November 2013

Assuming that salivary immunity to indigenous microorganisms could develop, we assessed antibacterial reactivities of natural salivary antibodies in specific pathogen‐free inbred mice. An ELISA was set up, using whole bacterial cells, to map reactivities of salivary IgA and IgG which accounted respectively for 91% and 8.7% of salivary Ig's in the BALB/c mouse. Representative strains of seven species from three genera (Lactobacilli, Staphylococci, and Streptococci), including major and minor components of the murine oral flora (38, 43, and 8%, respectively), were used to determine the presence and level of specific antibodies in individual saliva. It was verified that naturally occurring IgA antibodies can display diverse antibacterial reactivities. A characteristic profile emerged for salivary IgA where antibodies to Streptococcus faecalis predominate. Natural salivary IgG antibodies did not show the same reactivity pattern as IgA, anti‐Lactobacilli and anti‐Staphylococci reactivities being much less frequent in the salivary IgG repertoire. However, antibodies to S. faecalis occurred at the same high frequency for both isotypes (62–70% of the samples). Besides being species‐specific, antibacterial reactivities were also found to be strain‐specific. Broad variations in antibacterial titers were detected among individual mice under standardized experimental conditions. Present data thus suggest that the dynamics of salivary antibody production in the mouse reflect a differential natural sensitization of the secretory (IgA) versus the systemic (IgG) immune systems by distinct populations of indigenous bacteria.
Comparison of Wheat Germ Agglutinin‐ and Phorbol Myristate Acetate‐Mediated Triggering for Macrophage H2O2 Release
Authors: Tomioka Haruaki , Saito Hajime
Published: November 2013

Murine peritoneal macrophages (Møs) elicited by a single injection of zymosan A showed a higher responsiveness to the wheat germ agglutinin (WGA)‐ mediated triggering for H2O2 release than Møs activated by double injections of the agent. On the contrary, the response to phorbol myristate acetate(PMA)‐ mediated triggering was higher in the latter Møs than in the case of the former Møs. Furthermore, PMA‐triggered Mø H2O2 release was found to be inhibited by sarcoma 180 tumor cell‐derived proteinaceous factor in a much more marked fashion than the WGA‐triggered H2O2 release. These results indicate some significant differences between the cellular mechanisms of the WGA‐ and PMA‐triggering for Mø oxidative burst. On the other hand, microfilament‐inhibitors (cytochalasins B and E) and serine protease‐inhibitors (tosyl‐l‐lysine‐chloromethyl ketone and tosylamido‐2‐phenylethyl‐chloromethyl ketone), but not microtubule‐disrupting agents (colchicine, vinblastine, and vincristine), suppressed both the WGA‐ and PMA‐triggerings for Mø H2O2 release to a similar degree, thereby indicating that the WGA‐ and PMA‐triggerings for Mø oxidative burst have a common process which is dependent on microfilament and serine protease functions. In relation to this, the WGA‐ and PMA‐triggerings for Mø spreading were also depressed by microfilament‐ and serine protease‐inhibitors but not by microtubule‐inhibitors, indicating a participation of common membrane functions in the signal transduction in cases of Mø oxidative burst, and cell spreading induced by the WGA‐ as well as PMA‐triggering.
Mechanisms of Acquired Resistance in Mouse Typhoid
Authors: Chander Ramesh , Lewis Norman Francis
Published: November 2013

A polyvalent radiovaccine of Salmonella was shown to induce protective immunity in mice. The results revealed that the immunized mice progressively eliminated the challenged organisms and no Salmonella could be isolated after a period of 21 days. In contrast, Salmonella grew in the control mice and reached levels of 108 to 109 cfu/spleen resulting in the death of animals. Sera from both control and the immune mice were found to lack bactericidal activity. It was further observed that the vaccine induced delayed type of hypersensitivity and that antibody production as measured by bacterial agglutination and passive hemagglutination were low in response to the vaccine. However, the phagocytic activity of the reticuloendothelial system was considerably enhanced by the immunization. The results of experiments with immunosuppressed mice suggested the role of thymus derived (T) lymphocytes in the protection.
17K‐Spore Coat Protein Antigen in Sporulating Cells of Bacillus megaterium ATCC 19213
Authors: El‐Belbasi Hussein I. , Imagawa Masayoshi , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

Using immunological techniques, we studied the behavior of spore coat protein during sporulation of Bacillus megaterium ATCC 19213. Antibody specific to the main coat protein of 17,000 daltons was prepared and used to demonstrate that the spore coat protein was synthesized and deposited at a later stage during sporulation.
Immunoelectron Microscopic Localization of Mammary Tumor Virus (MTV) Antigens in Normal and Cancerous Mammary Glands of Mice
Authors: Tsubura Airo , Shikata Nobuaki , Hilgers Jo , Zotter Stefan , Tanaka Harutaka , Morii Sotokichi
Published: November 2013

Peroxidase‐labeled Fab' fragments of rabbit antisera against gp52 (major envelope protein) and A‐particles of mammary tumor virus (MTV) were prepared and used for investigation by immunoelectron microscopy of the replication cycle of MTV‐specific envelope and core antigens in normal and malignant mammary gland cells of female mice. The specificity of the antisera was proven by absorption tests and lack of reactivity to MTV‐free mammary tissues. Periodate‐lysine‐paraformaldehyde (PLP) fixation sufficiently preserved the antigenicity of gp52, while Zamboni's fixative was useful to preserve the core antigen. Saponin pretreatment was necessary to reveal the intracellular antigen of A particles but had no influence on gp52. In addition to its presence at the envelope of B particles, gp52 was clearly associated with the biomembrane system, including the nuclear membrane, endoplasmic reticulum, Golgi apparatus and plasma membrane independent of the presence of virus particles. In mammary tumors, a significant level of gp52 antigen was often expressed on the entire cell surface membrane. In contrast, it was localized only to the apical plasma membrane in normal mammary gland cells. A particle antigens were confined to the intracytoplasmic A particles, usually visible as clusters, and to the inner part of B particles. These ultrastructural findings support the available biochemical data on the morphogenesis of MTV particles.
The Immune System Response to Campylobacter Infection
Authors: Melamed Aizic , Zakuth Vera , Schwartz David , Spirer Zvi
Published: November 2013

Campylobacter may be one of the most common causes of bacterial gastroenteritis (GE) in children. It has recently been suggested that it is one of the bacterial pathogens most likely to infect immune‐compromised children, and it may facilitate colonization of enteric pathogens. The immune system response was studied in 12 children with Campylobacter fetus subspecies jejuni (CBJ) infections. Serum concentrations of IgA, IgM, and IgG were analyzed using a Beckman auto‐analyzer. Sera specific Ab to CBJ were tested with CBJ specific enzyme‐linked immunosorbant assay (ELISA). Mitogen stimulation of lymphocytes was performed to three lectins: Con A, PWM, and PHA. The lymphocyte blast transformation to Campylobacter was studied using the Campylobacter antigen. T‐cell subsets were studied using the monoclonal antibodies Leu 2, 3, and 4 (Becton Dickinson). Chemotaxis was measured in modified Boyden chambers; chemotactic stimulants were the Formyl Met Leu Phe, Campylobacter antigen virion, and E. coli 0111 B. Immunoglobulins were normal in nine cases and abnormal in two children previously diagnosed as agammaglobulinemic and one diagnosed as hypoagammaglobulinemic. Specific serum Ab level was significantly higher in the CBJ group, except in the agammaglobulinemic group. Stimulation indices to mitogens and monoclonal subset were in the normal range. The blastogenic transformation to CBJ Ag was decreased compared to normal lectins, and positive and high compared to controls. The chemotactic activity to campylobacter Ag was decreased in comparison to other stimulants. Most CBJ infections are self‐limiting due to a normal immune response and collaboration of all cellular limbs. When, however, the immune response is disturbed, we may find a prolonged and complicated course of CBJ.
Role of Pili in the Pathogenesis of Pseudomonas aeruginosa Burn Infection
Authors: Sato Hisaaki , Okinaga Kota , Saito Hiroshi
Published: November 2013

The present study using three isogenic mutants (F+P−, F−P+, F−P−) of Pseudomonas aeruginosa indicates that the presence of pili enhances the virulence of the organisms in experimental P. aeruginosa burn infection of mice. The 50% lethal dose (LD50) value for burned mice inoculated with non‐piliated (P−) mutant was at least ten times higher than those inoculated with piliated (P+) bacteria. Meanwhile the LD50 value for burned mice inoculated with non‐flagellated (F−) mutant was at least 105 times higher than those inoculated with flagellated (F+) bacteria. At 24 hr after inoculation, the bacterial counts in burned skin of mice inoculated with P+ bacteria were ten times higher than those inoculated with P− bacteria; and at 48 hr the bacterial counts became a hundred times higher in the former mice than the latter. At 24 hr after inoculation, P+ bacteria were isolated from blood, liver (F+P+), lung (F+P+), and kidney, while P− bacteria were not present in these tissues. And at 48 hr after inoculation, P+ bacteria were isolated from all tissues, while P− bacteria were isolated from some sites only. These results suggested that pili and flagella each play an important role as virulence factors independently, and that pili‐mediated enhancement of virulence of P. aeruginosa was attributed to pili‐mediated enhanced colonization of the organisms at the burned skin surfaces.
Small‐Scale DNA Preparation for Rapid Genetic Identification of Campylobacter Species without Radioisotope
Authors: Ezaki Takayuki , Takeuchi Noriko , Liu Shulin , Kai Akemi , Yamamoto Hiroaki , Yabuuchi Eiko
Published: November 2013

A simplified and rapid genetic identification method for Campylobacter species without radioisotope was established. Three different amounts of DNA (200, 50, and 12.5 ng) extracted from each type strain of Campylobacter species with standard Marmur's procedure were spotted on a nitrocellulose filter. DNA obtained from one ml bacterial suspension at a concentration of McFarland standard turbidity No. 1 of Campylobacter fetus, C. jejuni, C. coli, and C. pylori isolates were sufficiently labeled with photo‐biotin within 15 min and clearly hybridized with the type strain of the corresponding species within four to six hours. Hybridized spots were visualized with alkaline‐phosphatase‐conjugated streptavidin color‐detection method. The reaction was usually stopped within 30 min. Atypical clinical isolates such as a nitrate‐negative C. jejuni, two nalidixic acid‐resistant C. jejuni, and two strains of C. fetus able to grow at 42 C, which were tentatively identified as such, were definitely identified by the simplified DNA hybridization method presented here. This method will be applicable routinely for the definite identification of atypical strains of Campylobacter species and other gram‐negative bacteria difficult to identify biochemically.
In vitro Hexagonal Assembly of R‐Form Lipopolysaccharides: Effect of pH on the Mg+2‐Mediated Hexagonal Assembly
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Ito Hideo , Naito Setsuko
Published: November 2013

The R‐form lipopolysaccharide (LPS) from Escherichia coli K‐12, from which cationic material had been removed by electrodialysis and the pH of which had fallen to 3.6, formed a rough hexagonal lattice structure with the lattice constant of about 19 nm. The rough hexagonal structure was maintained in buffers at pH 5 or lower but disintegrated into the ribbon‐like structures in buffers at pH 6 or higher. However, in the presence of 10 mm Mg2+, the hexagonal lattice structure was not disintegrated even at alkaline pH levels but conversely it became more dense. At pH 8.3 to 8.9, the hexagonal lattice structure with the shortest lattice constant (15 nm) was formed. The same optimal pH levels were obtained for formation of the dense hexagonal lattice structure (lattice constant, 14 to 15 nm) by the electrodialyzed LPS from Klebsiella pneumoniae strain LEN‐111 (O3‐:K1‐). The ability of Mg2+ to induce formation of the dense hexagonal lattice structure of the K‐12 LPS depends upon the presence of buffers showing the optimal pH levels, since a very high concentration of Mg2+ such as 500 mm was required for the lattice formation in distilled water. The amount of the magnesium bound to the K‐12 LPS did not significantly differ throughout the pH range of 3 to 9. Therefore, the optimal pH range is another essential factor for formation of the dense hexagonal lattice structure of the LPS in addition to binding of the magnesium to the LPS.
Purification and Antigenic Analysis of Flagella of Campylobacter jejuni
Authors: Ueki Yuji , Fujimoto Shuji , Umeda Akiko , Amako Kazunobu
Published: November 2013

The flagella of Campylobacter jejuni strain FUM158432 were purified and a flagellin preparation consisting of only a single peptide of 63,000 daltons was obtained. The peptide of 92,000 daltons usually associated with a flagellar preparation was shown to be a peptide derived from the hook region. Antiserum was prepared by immunizing a rabbit with the flagellin preparation. The reaction of the antiserum was found to be highly specific for the flagellar filament by immunoelectron microscopy and for flagellin peptide by the immunoblotting method. Seventeen of 23 clinically isolated strains of C. jejuni reacted with this antiserum but the other six strains did not, indicating the existence of antigenic variation of the flagella of C. jejuni. The flagella of a few strains of C. coli also reacted with this antiserum.
Effect of Wheat Germ Agglutinin on T Lymphocyte Activation
Authors: Kawakami Kazuyoshi , Yamamoto Yusei , Onoue Kaoru
Published: November 2013

Wheat germ agglutinin (WGA) is low mitogenic or nonmitogenic for human T lymphocytes and inhibits phytohemagglutinin (PHA)‐induced mitotic response of the lymphocytes. In this study, the effect of WGA was analyzed in terms of interleukin 2 (IL2) production, expression of IL2 receptor, and IL2 responsiveness of the T lymphocytes. WGA as well as PHA could induce IL2 mRNA and IL2 production and also elevate cytoplasmic free Ca2+ concentration. The IL2 production was reduced by inhibitors of calmodulin and protein kinase C. The IL2 receptor (Tac) expression was induced at about 20% of the lymphocytes by WGA and the expression induced by PHA was not blocked by the addition of WGA. The lymphocytes precultured with WGA for 3 days could proliferate by the addition of IL2 after removal of WGA. The IL2‐dependent proliferation of PHA‐blasts was blocked by the addition of WGA. These results indicate that WGA inhibits T lymphocyte proliferation by inhibiting the responsiveness of the lymphocytes to IL2 but not by interfering with IL2 production and IL2 receptor expression.
Effect of Rickettsial Antigen‐Specific T Cell Line on the Interaction of Rickettsia tsutsugamushi with Macrophages
Authors: Kodama Kazuya , Yasukawa Masaki , Kobayashi Yuzuru
Published: November 2013

The effect of rickettsial antigen‐specific T cell line on macrophages infected with Rickettsia tsutsugamushi was studied. It was suggested that the T cells could produce a factor which acts cytotoxically and selectively on infected macrophages.
Mechanism of Streptomycin Resistance in Leptospira biflexa Strain Urawa
Authors: Fukunaga Masahito , Mifuchi Ichiji
Published: November 2013

The mechanism of streptomycin resistance of Leptospira biflexa was investigated. A streptomycin‐resistant mutant of Leptospira showed cross‐resistance to dihydrostreptomycin but not to other antibiotics. Enzymatic inactivation of the drug could not be demonstrated in this mutant. Protein synthesis on the ribosomes from the mutant was insensitive to streptomycin. These results suggest that ribosomal resistance is the reason for streptomycin resistance in Leptospira biflexa.
The Isolation of Mycobacterium avium Complex from Soil, Water, and Dusts
Authors: Ichiyama Satoshi , Shimokata Kaoru , Tsukamura Michio
Published: November 2013

Previously, it was difficult to isolate the Mycobacterium avium complex from soil, water, and dusts, because rapidly growing mycobacteria always overgrew slowly growing ones. We used Ogawa egg medium containing both ethambutol and ofloxacin, which inhibit the nonpathogenic slowly growing mycobacteria and most rapidly growing mycobacteria, respectively, as an aid to screen for pathogenic slowly growing mycobacteria; we could thereby isolate a number of the M. avium complex and M. scrofulaceum strains from soil, water, and dusts in this country.
Polyclonal B‐Cell Stimulative and Immunosuppressive Activities at Different Developmental Stages of Trypanosoma gambiense
Authors: Oka Mikio , Yabu Yoshisada , Ito Yoshihiro , Takayanagi Tan
Published: November 2013

Bloodstream trypomastigote and cultured procyclic (insect midgut) forms of a monomophic strain of Trypanosoma gambiense were tested for their abilities to induce polyclonal B‐cell activation (PBA) and immunosuppression (IS) in mice. Injection of a cell homogenate of bloodstream trypomastigotes induced both PBA and IS, while neither PBA nor IS was observed in mice injected with a cell homogenate of cultured procyclics. The results indicate that the substance(s) inducing PBA or IS is related to the developmental stage of the parasites.
Enhancement of Host Resistance against Listeria Infection by Lactobacillus casei: Efficacy of Cell Wall Preparation of Lactobacillus casei
Authors: Sato Katsumasa , Saito Hajime , Tomioka Haruaki , Yokokura Teruo
Published: November 2013

Cell wall, cytoplasm, polysaccharide, and peptidoglycan fractions prepared from Lactobacillus casei, L. plantarum, and L. acidophilus were examined for their efficacies to enhance resistance of host mice against Listeria monocytogenes infection. Intraperitoneal injections of those cellular fractions of L. casei led to elicitation of inflammatory cells in the peritoneal cavity and the efficacy was highest in the case of peptidoglycan. Macrophage ratio in the resultant peritoneal exudate cells was also highest in mice given peptidoglycan. Macrophages induced with cell wall fraction of L. casei showed the most potent phorbol myristate acetate (PMA)‐triggered respiratory burst (chemiluminescence and O2− production determined on the basis of nitroblue tetrazolium reduction) followed by those elicited with peptidoglycan. All the macrophages induced with cell wall of L. casei (two strains) and L. acidophilus enhanced O2− production in response to PMA but L. plantarum did not enhance O2−‐producing ability in such a manner. The L. casei‐cell wall also enhanced in vitro listericidal activity of mouse peritoneal macrophages, but such an activity was not noted in the case of L. acdiophilus‐cell wall. When mice were intravenously given the cellular fractions 7 or 13 days before L. monocytogenes infection, cell wall fractions of L. casei caused the most potent protective activity. A weak protective activity was also found in peptidoglycan of L. casei. Therefore, the protective action of L. casei against L. monocytogenes infection in host mice may be attributed to cell wall compounds and partially to the peptidoglycan moiety.
Characteristics of Mouse Lymphoid Cells Proliferating in vitro by Recombinant Human Interleukin 2 (TGP‐3): Comparison between Normal and Nude Mice
Authors: Hinuma Shuji , Naruo Ken‐Ichi , Shiho Osamu , Tsukamoto Kyozo
Published: November 2013

Various lymphoid cells obtained from BALB/c and BALB/c nu/nu mice were cultured in vitro with recombinant human interleukin 2 (rIL 2), and the characteristics of responder cells to rIL 2 were analyzed. Spleen cells, lymph node cells, and thymocytes except for bone marrow cells obtained from BALB/c mice remarkably proliferated in response to rIL 2. On the other hand, among lymphoid cells obtained from BALB/c nu/nu mice, only lymph node cells showed significant proliferation by rIL 2. Flow cytometric analyses revealed that mainly two types of lymphoid cells were proliferating in response to rIL 2 in BALB/c mice, i.e., Thy 1+, Lyt 1−, Lyt 2− and Thy 1+, Lyt 1−, Lyt 2+ cells. On the other hand, most of the proliferating cells were Thy 1+, Lyt 1−, Lyt 2− cells in BALB/c nu/nu mice. Treatment with various antibodies plus complement revealed that the majority of IL 2‐responsive cells in BALB/c mice were Thy 1+, Lyt 1+, and Lyt 2+, although a minor part of them were Thy 1−, Lyt 1−, and Lyt 2−. On the other hand, a predominant type of the IL 2‐responsive cells in BALB/c nu/nu mice were Thy 1−, Lyt 1−, and Lyt 2−, though some were Thy 1+. Nonspecific killer activity against tumor cells increased to variable extents in all of the lymphoid cells of both strains after culture with rIL 2. Our results indicate that mouse responder cells to rIL 2 have the following characteristics. First, the responder cells exist abundantly among spleen, lymph nodes, and thymus in normal mice, though their cell lineages are heterogeneous; one is of T cell lineage and the other of natural killer (NK) cell lineage. Second, nude mice are defective in the responder cells of T cell lineage but not of NK cell lineage. Moreover, the responder cells in nude mice predominantly accumulate in the lymph nodes but not other lymphoid organs.
Inhibition of Macromolecular Synthesis in Cultured Macrophages by Pseudomonas pseudomallei Exotoxin
Authors: Mohamed Rahmah , Nathan Sheila , Embi Noor , Razak Nyonya , Ismail Ghazally
Published: November 2013

Pseudomonas pseudomallei exotoxin was found to be a potent inhibitor of protein and DNA synthesis in cultured macrophages. Inhibition of DNA synthesis occurred at toxin concentrations as low as 1–2 μg/ml and inhibition of 3H‐thymidine uptake was almost complete at concentrations of 8 μg/ml or more. A close correlation between cell damage and inhibition by DNA synthesis was observed. For protein synthesis, inhibition was obtained at much lower doses (0.06–2.0 μg/ml) of the toxin. At similar toxin concentrations, DNA synthesis was marginally affected. Further, it was shown that protein synthesis inhibition occurred almost immediately after incubation, reaching its maximal inhibitory effect of 70% after 6 hr. DNA synthesis, however, was minimally affected by a similar toxin concentration even after 10 hr of incubation. The inhibition of macromolecular synthesis in macrophages by P. pseudomallei exotoxin may be relevant to its modulatory effect on the host defense mechanism.
Electron Microscopic Study of a Herpes‐Type Virus Isolated from an Infant with Exanthem Subitum
Authors: Yoshida Mariko , Uno Fumio , Bai Zeng Liang , Yamada Masao , Nii Shiro , Sata Tetsutaro , Kurata Takeshi , Yamanishi Koichi , Takahashi Michiaki
Published: November 2013

Human lymphocytes and MT4 cells infected with a virus isolated from a patient with exanthem subitum were examined by transmission and scanning electron microscopy. The most striking characteristic of the ultrastructure of this herpes‐type virus was that nucleocapsids located outside the nucleus were each coated distinctly with a tegument of moderate electron density. Tubular structures formed due to some mistakes in the viral assembly were also detected in the nucleus. Morphological differentiation of this virus from the other human herpesviruses was discussed. From these observations it was concluded that this virus has the same ultrastructural characteristics as HBLV (HHV‐6).
Prevalence of Small Round Structured Virus Infections in Acute Gastroenteritis Outbreaks in Tokyo
Authors: Sekine Seiji , Okada Shojiro , Hayashi Yukinao , Ando Tamie , Terayama Takeshi , Yabuuchi Kiyoshi , Miki Takashi , Ohashi Makoto
Published: November 2013

During the three‐year period from 1984 to 1987, 506 acute gastroenteritis outbreaks involving 14,383 patients were reported to the Bureau of Public Health, Tokyo Metropolitan Government. Eighty (4,324 patients) of 150 outbreaks (4,860 patients) from which etiologic agents were not identified were subjected to virological investigation. Spherical particles of 28–32 nm in diameter with capsomere‐like structures on the surface were detected in patients' stool specimens. Buoyant density of the particles appeared to be 1.36 to 1.40 g/ml in CsCl. Seroconversion to the particles was observed in patients by immune electron microscopy. From these observations, we concluded that the detected particles were members of small round structured virus (SRSV), and that they were implicated in the etiologically ill‐defined outbreaks encountered. Prevalence of SRSV infections in these outbreaks was examined by electron microscopy. SRSV was positive in 83.8% of the outbreaks, and 96.4% of the cases. SRSV‐positive outbreaks usually occurred during winter in contrast to bacterial outbreaks which often occurred in the summer season. Of 80 outbreaks examined, 53 were associated with the ingestion of oysters, and the remaining 27 mostly with food other than oysters. Oyster‐associated outbreaks usually occurred on a small scale, while unassociated ones on diverse scales ranged from family clusters to large outbreaks.
Strain Differences in the Early Development of the Thymus‐Dependent Cells: Precocity of T Lineage Cells in AKR Mice as Compared to Those in C3H Mice
Authors: Katsume Chikako , Fernandes Gabriel , Iwabuchi Kazuya , Ogasawara Kazumasa , Gotohda Toshihiko , Good Robert A. , Onoé Kazunori
Published: November 2013

Early development of T lineage cells were compared between AKR and C3H mice by using two experimental strategies—neonatal thymectomy (NTx) and bone marrow transplantation (BMT)—between these two strains of mice. After NTx, AKR mice developed less wasting disease and showed better maintenance of several T cell functions. In addition, the response of neonatal spleen cells to PHA and ConA was much greater in AKR mice than in C3H mice. Further, when AKR mice were used as recipients of BMT, cell numbers recovered from thymuses between 2 and 7 weeks after reconstitution were consistently much greater (about 10 times greater) than those from chimeras where C3H mice were used as recipients, regardless of the donor strains of bone marrow cells. However, 4 weeks after BMT the proliferative responses to ConA were consistently higher in the donor‐derived thymocytes from chimeras where AKR mice were used as bone marrow donors than in those from chimeras in which C3H were donors. The present findings suggest that these differences may be attributed to characteristics of recipient microenvironment (e.g., thymic stroma) which maintain developing thymocytes and supply them to the peripheral lymphoid tissue. Alternatively the differences may to some degree also be attributable to characteristics of the thymic progenitors themselves, which may determine the rates of maturation of thymocyte functions.
The Number of Large Ribosomal RNA Genes in Leptospira interrogans and Leptospira biflexa
Authors: Fukunaga Masahito , Mifuchi Ichiji
Published: November 2013

We determined the number of large ribosomal RNA genes in five strains of Leptospira by hybridization of 15 restriction endonuclease digests of genomic DNA to the [32P]‐labeled fragment of 23s rRNA gene. Almost all the restriction gels gave two radioactive bands. The conclusion from these results is that there are at least two rRNA genes in these leptospiral strains. Furthermore, the hybridization patterns of L. icterohaemorrhagiae strains Ictero No. I and RGA are almost identical. The number of rRNA genes and taxonomic relationships of these leptospires were discussed.
Delayed‐Type Hypersensitivity Skin Reaction to HBsAg and Immunohistopathologic Study of Liver in Patients with Acute Type B Viral Hepatitis
Authors: Nagafuchi Seiho , Kashiwagi Seizaburo , Tsuji Yuji , Miyata Yasuji , Iwakiri Ryuichi , Takeshita Morishige , Niho Yoshiyuki , Kikuchi Masahiro
Published: November 2013

In an attempt to clarify the role of cellular immunity in the pathogenesis of acute type B viral hepatitis (AHB), we studied delayed‐type hypersensitivity (DTH) skin reaction to hepatitis B surface antigen (HBsAg) and immunohistopathology of livers in patients with AHB. DTH skin reaction to HBsAg developed early in the convalescent phase in all 14 patients with AHB. In contrast, the production of anti‐HBs was significantly delayed in these patients, compared with healthy controls immunized with HB vaccine intradermally (P< 0.001). These observations suggest that DTH to HBsAg but not anti‐HBs may be associated with recovery from AHB. In the biopsied livers obtained from patients with AHB, the proliferation of Kupffer cells was extensive and immunohistopathologic studies revealed an accumulation of CD‐4 positive lymphocytes in the portal area, a finding which suggested a DTH reaction in the liver with AHB. CD‐8 positive cells had infiltrated the lobule and made contact with affected hepatocytes, thereby indicating that a cytotoxic T cell response is involved in damaging of the infected cells. All these observations taken together, we propose that not only a cytotoxic T cell response but also a DTH reaction may be involved in the pathogenesis of AHB.
Syncytium‐Inducing Capacity of Human Immunodeficiency Virus (HIV): Analysis by the Use of Cloned Viruses
Authors: El‐Farrash Mohamed A. , Harada Shinji
Published: November 2013

The marked cytopathic effects of human immunodeficiency virus (HIV) for susceptible cells are caused mainly by fusion between cells expressing viral envelope glycoproteins and cells expressing CD4 molecule. In this study, we tested the ability of different clones of HIV to induce syncytia in CD4‐positive cells. We have reported marked difference in syncytium‐inducing capacity of 2 clones of human T lymphotropic virus type III (HTLV‐IIIB) isolate despite no detectable difference in expression of viral glycoprotein (gpl20). This difference in syncytium induction could be explained by the difference detected in their infectivity and binding activities to CD4‐positive cells. Meanwhile we reported difference in syncytium‐inducing capacity of 2 clones of lymphadenopathy associated virus (LAV1) isolate parallel to the different amounts of gpl20 and other viral proteins expressed by these 2 clones. These results suggest that viral factors like infectivity and binding affinity of the virus to the susceptible cells and the amount of viral gpl20 expressed by the infected cells may interact in a complex manner affecting fusion activity and syncytium induction in CD4‐positive cells.
Hemolytic Vibrio cholerae O1 That Is Sensitive to Mukerjee's Cholera Phage IV and the Phage Produced by the Hemolytic Vibrio Lysogenized with the Infection of Mukerjee's Cholera Phage IV
Authors: Iwanaga Masaaki , Nakamura Satoshi , Kuyyakanond Thicumporn
Published: November 2013

Strains of hemolytic Vibrio cholerae Ol (El Tor vibrio) which are sensitive to Mukerjee's cholera phage group IV were isolated from cholera patients in North‐East Thailand in 1986. Plaques of the phage on these hemolytic V. cholerae Ol were usually translucent but almost transparent on some strains, just like the plaques on non‐hemolytic V. cholerae Ol (classical vibrio). These hemolytic V. cholerae Ol were lysogenized with the infection of cholera phage IV, and the lysogenized strains produced phage different from cholera phage IV. These hemolytic strains were classified into Cured type in prophage typing of V. cholerae Ol, El Tor, because they were also lysogenized with Kappa phage and were hemolytic. When Cured‐type V. cholerae Ol, El Tor previously isolated in various countries were examined for the sensitivity to cholera phage IV, some of the isolates were sensitive.
Chemotaxonomic Study of Vibrio cholerae Bio‐Serogroup Hakata on the Basis of the Sugar Composition of the Polysaccharide Portion of Its Lipopolysaccharides
Authors: Kondo Seiichi , Iguchi Takehiro , Haishima Yuji , Isshiki Yasunori , Hisatsune Kazuhito
Published: November 2013

A chemotaxonomic study was carried out on 31 strains of non‐O1 Vibrio cholerae bio‐serogroup Hakata, isolated in Japan, which possesses the Inaba antigen C of O1 V. cholerae. On the basis of the compositional sugar pattern of the polysaccharide portion of their lipopolysaccharides, the 23 strains isolated from the environment were separated into two groups, one (20 strains) containing mannose, glucose, fructose, L‐glycero‐D‐mannoheptose, glucosamine, perosamine, quinovosamine, and an unidentified amino sugar AS, and the other (3 strains) containing two additional sugars, galactose and a trace amount of galactosamine. All of the eight strains isolated from imported seafoods belonged to the former group.
Morphological Detection of Filipin‐Sterol Complexes in the Cytoplasmic Membrane of Staphylococcal L‐Form
Authors: Nishiyama Yayoi , Yamaguchi Hideyo
Published: November 2013

Filipin, a sterol‐specific antibiotic, and freeze‐fracture electron microscopy were used to study the presence and distribution of sterol in the cytoplasmic membrane of stable staphylococcal L‐form cells. Fixed cells were treated with filipin, and then observed by freeze‐fracture electron microscopy. Freeze‐fractured profiles of the L‐form cells treated with filipin demonstrated irregular distribution of protuberances or pits of 25–30 nm, representing filipin‐sterol complexes, on the protoplasmic fracture face (PF) and exoplasmic fracture face (EF) of the cytoplasmic membrane. In contrast, no such structure was detected in the filipin‐treated parent cells or protoplasts. The results suggest that some sterol molecules, which are usually not found in staphylococcal or other bacterial cells, emerged on the cytoplasmic membrane after the cells were converted to the stable L‐form.
Genotypic and Phenotypic Differentiation of Flavobacterium indologenes Yabuuchi et al 1983 from Flavobacterium gleum Holmes et al 1984
Authors: Yabuuchi Eiko , Hashimoto Yasuhiro , Ezaki Takayuki , Ido Yoshimi , Takeuchi Noriko
Published: November 2013

The type and eight strains of Flavobacterium indologenes were clearly differentiated from the type and two reference strains of Flavobacterium gleum by deoxyribonucleic acid‐deoxyribonucleic acid homology data and phenotypic characteristics. Phenotypic characteristics useful to differentiate the two species are presented.
Protective Effect of Recombinant Human Interleukin‐2 against Lethal Infection Caused by Klebsiella pneumoniae
Authors: Iizawa Yuji , Nakao Masafumi , Kondo Masahiro , Yamazaki Toshiyuki
Published: November 2013

The effects of recombinant human interleukin‐2 (rIL‐2) administered prophylactically on the death of CBA/J mice challenged with Klebsiella pneumoniae 27 intraperitoneally were examined. rIL‐2 administered subcutaneously at 20 μg per mouse for 7 days enhanced survival after a lethal challenge. The injection of anti‐asialo GM1 antibody did not influence the effect of rIL‐2. In mice given rIL‐2. the number of peritoneal macrophages increased, and the infiltration of polymorphonuclear leukocytes (PMN) into the peritoneal cavity after the bacterial challenge was enhanced. In addition, adoptive transfer of sera and peritoneal exudate cells (PEC), consisting of an approximately equal number of macrophages and PMN, obtained from mice given rIL‐2 enhanced resistance to a K. pneumoniae infection, compared with adoptive transfer of sera and PEC obtained from mice not given rIL‐2. These results indicate that rIL‐2 protects mice from a lethal challenge with K. pneumoniae. and suggest that the protective effect is due to an increase in the number of phagocytic cells and in the cooperative activity of the sera and the phagocytic cells.
Changes in Interleukin 1 Levels in Human Amniotic Fluid with Gestational Ages and Delivery
Authors: Tsunoda Hajime , Tamatani Takuya , Oomoto Yasukazu , Hirai Yoshikatsu , Kasahara Tadashi , Iwasaki Hirokazu , Onozaki Kikuo
Published: November 2013

In order to understand the role of interleukin 1 (IL‐1) in pregnancy, the amount of IL‐1 in normal human amniotic fluid (AF) from various gestational ages and delivery was measured using an ELISA. AF samples were divided into three groups of varying gestational ages. Group 1 of AF was collected by amniocentesis from gestational ages <24 weeks (n = 13). Group 2 was collected transvaginally during delivery following labor ≥36 weeks (n = 36). Group 3 was transabdominally collected from elective cesarean section without labor ≥36 weeks (n −8). IL‐1α was present in AF of early gestational age, 19.2±21.7 pg/ml, in group 1, and appeared to increase with gestational age, 63.4±50.1 pg/ml, in group 3. In contrast, IL‐1β was not detectable in either group 1 or 3. However, the concentration of IL‐1 in group 2 was extremely high (IL‐lα, 233.1±351.9 pg/ml; IL‐1β, 1,093.5±1,369.7 pg/ml) compared to the other groups. Moreover, these concentrations tended to increase with the duration of labor. IL‐1α and IL‐1β concentrations in AF were intimately related. These findings suggest that IL‐1 has some roles during pregnancy and especially during labor.
Isolation of Listeria monocytogenes from Raw Milk and Its Environment at Dairy Farms in Japan
Authors: Takai Shinji , Orii Fumie , Yasuda Kenzou , Inoue Satoshi , Tsubaki Shiro
Published: November 2013

The incidence of Listeria monocytogenes contamination in raw milk from farm bulk tanks and silage was investigated monthly at 20 dairy farms in Aomori prefecture, Japan, from January to July in 1989. Listeria sp. was isolated from one sample (5%) of raw milk collected in May. but L. monocytogenes was not isolated from raw milk. L. monocytogenes was isolated from two samples of silage (10%) collected in June. Silage, cow feces, sawdust bedding, dust and soil in these farms were tested for listerias, and we confirmed listerias contamination of the environment in the dairy farms.
Titer and Specificity of Autoantibody to β2‐Microglobulin in Sera from Patients with Rheumatic Disease
Authors: Yokochi Takashi , Yanagawa Akira , Nakagawa Takemasa , Mizushima Yutaka
Published: November 2013

Sera from patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and mixed connective tissue disease (MCTD) possessed higher titer of antibody to human β2‐microglobulin (β2m) than those from healthy controls and patients with Behçet's disease in the enzyme‐linked immunosorbent assay. It was also confirmed by the immunoprecipitation method. Anti‐β2m antibody in sera from those patients immunoprecipitated free β2m but not β2m in association with major histocompatibility complex class I antigen heavy chain. It was suggested that anti‐β2m antibody in sera from either patients or healthy controls might be directed mainly against free β2m. The relationship between the anti‐β2m antibody and anti‐lymphocytotoxic antibody found in those patients is discussed.
Participation of Calcium Ion on Depletion Mechanism of Liver Glycogen by Purified Glucocorticoid Antagonizing Factor Released in Blood during Endotoxemia
Authors: Sakaguchi Shuhei , Ibata Hiroshi , Yokota Katsushi
Published: November 2013

The present study was conducted by the use of purified glucocorticoid antagonizing factor (GAF) released in blood of endotoxemic mice to determine whether or not the factor (GAF and Ca2+) may play a possible role of mediator in depletion mechanism of liver glycogen in endotoxemia. The liver glycogen level in 2 hr after injection with GAF plus cortisone‐treated mice was markedly lower than that in cortisone alone‐treated mice. However, the administration of trifluoperazine or verapamil markedly increased glycogen levels in liver of GAF plus cortisone‐injected mice. On the other hand, when the mice fed a calcium‐free diet were injected with GAF plus cortisone, there was merely a significant difference in liver glycogen level as compared to cortisone alone‐treated mice. The level of Ca24+ in liver cytosol fraction in cortisone‐treated mice was higher 2 hr after GAF injection than that in the cortisone alone‐treated one. The phosphorylase a activity in liver 2 hr after injection of GAF plus cortisone did not show a significant difference as compared to that in mice treated with cortisone alone. However, the activity ratio of glycogen synthase enzyme (synthase I/synthase I + D) was decreased in GAF plus cortisone‐treated mice as compared to that in cortisone alone‐treated mice. These findings suggest that there are participations of Ca2+ and mediator GAF released from reticuloendothelial system (RES) macrophages in glucoregulation of endotoxemia. Thus, it may be speculated that intracellular Ca2+ may mediate glycogenesis rather than glycogenolysis in the depletion mechanism of liver glycogen during GAF‐poisoning.