bluuebunny/crossref_metadata_2025 huggingface dataset:
Relation between D‐Glucose and L‐ and D‐Alanine in the Initiation of Germination of Bacillus subtilis Spore
Authors: Yasuda Yoko , Tochikubo Kunio
Published: November 2013

The rate of L‐alanine‐initiated germination of Bacillus subtilis spore was measured by both loss of heat resistance and loss of turbidity, and the effect of glucose on the germination response to a wide range of concentrations of the germinant was analyzed in the presence and absence of D‐alanine, an inhibitor. Glucose stimulated L‐alanine germination by means of a cooperative effect: glucose increased the affinity of L‐alanine by about 3‐fold and the rate of germination by about 1.3‐fold. However, glucose had little effect on the binding affinity of D‐alanine. The apparent binding constant of L‐alanine to the spore, which was determined by the next measurable event in the trigger reaction, was 1.2 × 10−5, that of D‐alanine was 6 × 10−6, and that of glucose was 5 × 10−5. The relation between the binding site for glucose and those for L‐ and D‐alanine on the spore is discussed. Effect of glucose analogs was also examined.
Interrelationship between Drug Resistance and Bacteriocinogeny of Clostridium perfringens
Authors: Miyoshi Yasushi , Higa Akiko
Published: November 2013

One hundred and eight strains of Clostridium perfringens were collected from polluted waters and tested for their drug resistance and bacteriocinogeny. Thirty of the strains were tetracycline resistant and 36 were bacteriocinogenic. Only one strain possessed both tetracycline resistance and bacteriocinogen.

Transfer experiments on tetracycline resistance and bacteriocinogeny were carried out with several selected strains among these isolates. Both tetracycline resistance and bacteriocinogeny were shown to be transferable by conjugation‐like procedures. Transfer experiments on these two properties revealed that there were two types of recipient strains among the isolates in regard to the acceptance of tetracycline resistance and bacteriocinogeny. In one type of the recipient strain, these two properties seemed to be incompatible although in the other, both tetracycline resistance and bacteriocinogeny were transferred by two‐step mating. Transcipients of this two‐step mating retransferred their tetracycline resistance but not bacteriocinogeny to another recipient strain. The existence of an incompatibility‐like relationship between tetracycline resistance and bacteriocinogeny in one recipient strain, and the inability of these two properties to be co‐transferred from the two‐step transcipients, suggest a certain interrelationship between the factors responsible for tetracycline resistance and bacteriocinogeny.
Ecology of Non‐O 1 Vibrio cholerae in Toyama Prefecture
Authors: Kodama Hirohide , Gyobu Yotaku , Tokuman Naoko , Okada Itsuko , Uetake Hisao , Shimada Toshio , Sakazaki Riichi
Published: November 2013

The ecology of non‐O 1 Vibrio cholerae and Vibrio mimicus as causes of cholera‐like diarrhea or seafood‐associated gastroenteritis has been investigated in Toyama Prefecture since 1980. The relationship between biological or serological characteristics of the isolates and their enteropathogenicity is discussed.

Overall isolation rates from river water, sea water, and fish were 24.0, 59.5, and 33.7%, respectively, the isolation frequency being, in general, extremely high in the summer season, although the organisms were detected all year around in the case of sea water. Most isolates from river water were unable to grow on plates of TCBS agar to which colistin was added at a concentration of 1 μg/ml (CL‐TCBS). These strains quickly fermented cellobiose. O‐51 and O‐70 were the two most frequently detected serogroups among them and they did not show enteropathogenicity in the rabbit ileal loop (RIL) test. On the other hand, almost all isolates from sea water and fish as well as those from human diarrhea cases were able to grow on CL‐TCBS, but were unable to ferment cellobiose quickly. O‐36, O‐10, O‐6, O‐8, O‐39, and O‐26 were the dominant serogroups of these isolates, and some of them showed enteropathogenicity in the RIL test. Six out of 98 isolates from river water, 14 out of 116 from sea water, and 19 out of 112 from fish were classified as Vibrio mimicus. All of these strains were able to grow on CL‐TCBS and quickly fermented mannose but not cellobiose. I‐41 was the most common serogroup among them and some of these strains showed enteropathogenicity in the RIL test. Production of a cholera‐like enterotoxin among the isolates in Toyama Prefecture, if any, seemed to be poor.
Isolation of Immunogenic and Suppressogenic Glycoproteins from Adenovirus Type 12 Hamster Tumor Cells
Authors: Yamamoto Hiroshi
Published: November 2013

Two types of glycoproteins were isolated from the membrane fraction of adenovirus type 12 (Ad12) hamster tumor cells by recovering detergent‐solubilized glycoproteins using concanavalin A‐affinity chromatography and gel filtration. One of the glycoproteins consisted of a polypeptide of 130,000 daltons (130K) with a pI value of 4.7–5.1, and the other consisted of a polypeptide of 18,500 daltons (18.5K) with a pi value of 6.3–6.6. The glycoproteins were immunologically different. The 18.5K glycoprotein induced in vivo resistance to tumor growth and anti‐tumor cytotoxic T cells, while the 130K glycoprotein induced in vivo suppressor T cells which inhibited the activity of anti‐tumor cytotoxic T cells.
Fucoidan Blocks Macrophage Activation in an Inductive Phase but Promotes Macrophage Activation in an Effector Phase
Authors: Sugawara Isamu , Lee Kwok Choy
Published: November 2013

It has been suggested that some polysaccharides play important roles in immune responses. Therefore, we used various types of polysaccharides for analysis of macrophage‐mediated tumor cell killing.

We report here that fucoidan blocked macrophage activation occurs in an inductive phase but enhanced macrophage activation appears in an effector phase.
The Appearance and Characterization of Cyanide‐Resistant Respiration in the Fungus Candida albicans
Authors: Aoki Shigeji , Ito‐Kuwa Shoko
Published: November 2013

The respiration of yeast‐form cells of the dimorphic fungus Candida albicans became resistant to cyanide during aging treatment in the resting state. An alternative, cyanide‐resistant respiratory pathway was found to develop fully in cells aged at a concentration of 0.75 × 109/ml or more at 25 C, but did not appear at 5 C. Chloramphenicol did not prevent the appearance of the alternative respiratory pathway. The effects of inhibitors, salicylhydroxamic acid (SHAM) and disulfiram (tetraethylthiuram disulfide), on respiration of aged cells were examined, and results indicated that SHAM binds at a site on the alternative respiratory pathway whereas disulfiram binds at two sites, one on the conventional respiratory pathway and the other on the alternative pathway. Thus, SHAM is a more selective inhibitor of the alternative respiration of C. albicans cells. SHAM‐titration of the alternative respiration revealed that less than 10% of the maximal activity of the alternative respiratory pathway was utilized under normal conditions, indicating that the alternative respiratory pathway makes a small contribution to the total respiration. It was therefore concluded that the alternative, cyanide‐resistant respiratory pathway operates fully when the cyanide‐sensitive, cytochrome pathway is blocked although aged cells possess both respiratory pathways.
Characterization of Generalized Transducing Phage φw39 Heteroimmune to Phage P1 in Escherichia coli W39
Authors: Yoshida Yoko , Mise Katsutoshi
Published: November 2013

Generalized transducing phage similar to phage P1 in Escherichia coli was isolated from E. coli W39, an antigenic test strain of the O121 group. This phage, designated φ5w39, was reciprocally heteroimmune to phages P1 and P7, but non‐reciprocally heteroimmune to phage D6. Transduction experiments using various R plasmids with different molecular weights suggested that phage φw39 could transduce at least 65 megadaltons DNA. As in the case of P1 prophage, φ5w39 prophage existed as a plasmid belonging to incompatibility group Y and carried a dnaB‐like function. The molecular weight of φ5w39 plasmid was nearly the same as that of plasmid, i.e., 58.6 megadaltons. Despite the pronounced structural and functional similarity of phages φw39 and P1, restriction cleavage patterns of their genomes differed considerably.
Growth of Measles and Subacute Sclerosing Panencephalitis Viruses in Human Neural Cell Lines
Authors: Sakaguchi Masahiro , Yoshikawa Yasuhiro , Yamanouchi Kazuya
Published: November 2013

Growth of cell‐free subacute sclerosing panencephalitis (SSPE) virus was compared with that of measles virus in three human neural cell lines: neuroblastoma, oligodendroglioma, and glioblastoma. The Edmonston strain of measles virus replicated in these neural cells as efficiently as in Vero cells. In contrast, the growth of the Mantooth strain of SSPE virus was suppressed moderately in neuroblastoma cells and markedly in oligodendroglioma and glioblastoma cells in spite of the induction of apparent cytopathic effects in these cells. Virus adsorption, defective interfering particles, interferon, and temperature sensitivity were not responsible for this low yield of SSPE virus in neural cell lines. Synthesis of viral proteins of SSPE virus was slower than that of measles virus in oligodendroglioma and glioblastoma cells. These results suggest that the slow rate of synthesis of viral proteins may be relevant to the low yield of SSPE virus in neural cells.
4‐Hydroxy‐3‐Nitrophenyl (NP) Acetyl‐Hapten Specific Lymphocyte Proliferation
Authors: Okuda Kenji , Saito Harukazu , Aihara Yukoh , Minami Mutsuhiko , Tadokoro Ichiro
Published: November 2013

Hapten specific T cell proliferation was induced in several strains of mice. When lymph node T cells from 4‐hydroxy‐3‐nitrophenyl acetyl‐keyhole lympet hemocyanin (NP‐KLH)‐primed mice were stimulated in vitro with NP‐polymer glutamic acid‐lysine‐phenyl alanine (NP‐GLφ) or NP‐ovalbumin (NP‐OVA), they displayed a good level of proliferative responses. It was observed that NP‐GLφ could induce NP‐hapten specific proliferation even with NP‐KLH lymphocytes from GLφ non‐responder strains.

NP‐KLH primed lymphocytes from C57BL/6 (H‐2b, Igh‐1b), CKB (H‐2k, Igh‐1b), CWB (H‐2b, Igh‐1b), and B10.BR (H‐2k, Igh‐1b) mice showed good proliferative responses to both 4‐hydroxy‐5‐iodo‐3‐nitrophenyl (NIP) acetyl‐GLφ and NIP‐OVA antigens. However, NP‐KLH primed lymphocytes from C3H/He (H‐2k, Igh‐1j) and C3H. SW (H‐2b, Igh‐1j) mice displayed poor proliferative responses to NIP‐GLφ and NIP‐OVA antigen. These results suggested that the gene coding for the NIP‐cross‐reaction might be mapped in the Ig heavy‐chain linked locus.
Ultrastructure of Klebsiella O3 Lipopolysaccharide Isolated from Culture Supernatant: Comparison with Other Lipopolysaccharides
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Naito Setsuko , Kuno Tsuneharu
Published: November 2013

Klebsiella O3 lipopolysaccharide (KO3 LPS) isolated from the culture supernatant, which was found to exhibit a very strong adjuvant activity in augmenting antibody response and delayed‐type hypersensitivity to protein antigens in mice, was examined by electron microscopy. When negatively stained with uranyl acetate or ammonium molybdate, the KO3 LPS was found to consist principally of flat ribbon‐like structures branching freely (average width 16 nm and average thickness 7 nm) and to contain a small proportion of spheres (diameter 20–50 nm), both structures covered with fine hairy structures (average length approximately 10 nm). When the polysaccharide of KO3 LPS was stained by the periodic acid‐thiosemicarbazide‐silver proteinate procedure, silver granules were deposited on the ribbon‐like structures and around the spheres, suggesting that the polysaccharide moiety is located on their surface and that the fine hairy structures consist of the polysaccharide moiety. Comparison by means of preparations stained with uranyl acetate or ammonium molybdate showed that KO3 LPS isolated from the culture supernatant has structural features in common with KO3 LPS isolated from bacterial cells, Escherichia coli O9 LPS isolated from the culture supernatant, and E. coli O127 LPS isolated from bacterial cells. On the basis of the present results, schematic representations of the common physical structure of LPS were drawn; the fine hairy structures attach to the wide surface of the flat ribbon‐like structures along their lateral margin.
Ultrastructure of Klebsiella O3 Lipopolysaccharide Isolated from Culture Supernatant: Structure of Various Uniform Salt Forms
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Naito Setsuko , Kuno Tsuneharu
Published: November 2013

Various uniform salt forms of Klebsiella O3 lipopolysaccharide (KO3 LPS) isolated from culture supernatant were prepared as follows. Basic materials present in KO3 LPS were rigorously removed by electrodialysis and the electrodialyzed KO3 LPS was neutralized with NaOH, KOH, NH4OH, Ca(OH)2, tris(hydroxymethyl)aminomethane, or triethylamine. The ultrastructure of the uniform salt forms of KO3 LPS was examined using preparations stained with uranyl acetate. The sodium, potassium, ammonium, and trisaminomethane salt forms were structurally very similar to the natural form of KO3 LPS which consisted of a mixture of flat ribbon‐like structures (average width of 16 nm and average thickness of 7 nm) and spheres with various diameters, both covered with fine hairy structures. When KO3 LPS was converted to the triethylamine salt form, the ribbon‐like structures were distrupted into very small granules (7–9 nm × 9–15 nm). The calcium salt form consisted of particles and rods of various sizes and ribbon‐like structures which were markedly extended (maximum width of 50 nm) and presented irregular shapes. When converted to the calcium salt form, the ribbon‐like structures were extended and eventually divided into particles and rods. For reasons still unknown, the sodium salt of KO3 LPS was mostly stained positively with uranyl acetate in contrast to the natural form and the other uniform salt forms which were always negatively stained. In the positively stained preparation of the sodium salt form, it was clearly shown that the ribbon‐like structures consisted of a bilayer.
Viral Pollution of the Rivers in Toyama City
Authors: Matsuura Kumiko , Hasegawa Sumiyo , Nakayama Takashi , Morita Osayuki , Uetake Hisao
Published: November 2013

Viral pollution of the river water in Toyama City was surveyed during the two‐year period from July 1979 to July 1981, and the ecology of viruses in the river water is discussed. Virus isolation from the river water samples, or from the water squeezed from cotton pads that were immersed in the stream for 3 days, was carried out by the “filter adsorption/elution” method.

River waters were found to be contaminated with various species of enteric viruses, that is, poliovirus, echovirus, coxsackievirus, adenovirus, and reovirus. Poliovirus was isolated during the period immediately after the oral administration of polio vaccine, and Coxsackie B virus was frequently isolated all year around. The enterovirus concentration in the river water was significantly high with a maximum of five plaque‐forming units of Coxsackie B2 virus per 250 ml.

The species and type distribution of enteroviruses isolated from the river water coincided well with that of viruses isolated from inhabitants of Toyama Prefecture, with the exception of reovirus which was the largest population of virus species in the river water.
Unique Nature of an Attenuated Strain of Tobacco Mosaic Virus: Autoregulation
Authors: Kiho Yukio , Nishiguchi Masamichi
Published: November 2013

An attenuated strain L11A of tobacco mosaic virus (TMV) multiplied like wild type strain L at an early stage of infection in tomato leaves. Four days after inoculation, however, multiplication of L11A was drastically reduced (autoregulation) compared with the constant multiplication of L. In mixed infections, L11A strongly inhibited the multiplication of homologous strain L. Experiments with cucumber mosaic virus (CMV) or tobacco plants revealed that the inhibitory mechanism of L11A is not host‐specific but virus‐specific, and the auto‐regulatory mechanism is effective only for TMV.

RNA synthesis in L11A infected leaves 4 days after inoculation was studied by polyacrylamide gel electrophoresis. Synthesis of TMV‐RNA and its replicative intermediate were strongly inhibited, whereas the replicative form of TMV‐RNA and ribosomal RNA were synthesized as in the case of L infection.

Synthesis of non‐coat‐protein was studied by the incorporation of radioactive histidine into subcellular fractions derived from leaves infected with L or L11A for 4 days. Different patterns of the two strains in protein synthesis were noted. At least three proteins were predominantly synthesized in L11A infection. One of them was observed in the mitochondria fraction. From its position in Polyacrylamide gel, it could be viral coded 165K protein which is considered to be involved in viral RNA replication.

These results suggest that the unique nature of attenuated virus L11A, i.e. autoregulation, resulted from the inhibitory mechanism of viral RNA synthesis due to overproduction of 165K protein and is quite distinct from interferon, intrinsic interference or interference by defective virus.
Immune Interferon Production by TH69, a Lyophilized Preparation of Streptococcus faecalis, in Murine Spleen Cell Cultures
Authors: Yamaguchi Takahiro , Kuroda Yoko , Saito Motoo , Ebina Takusaburo , Hoshino Fumihiko , Ishida Nakao
Published: November 2013

In mouse spleen cell cultures, TH69, a live Streptococcus faecalis (ATCC 31663) preparation, at a concentration of 20 μg/ml induced immune interferon (IFNγ) with molecular weight ranging from 20,000 to 40,000 daltons together with a small amount of IFNα/β.

By using nonsensitized mouse spleen cells, the fact that both T‐cells and macrophages are required for this IFN production was established. When these spleen cells were obtained from mice sensitized 12 days earlier with 4 mg of TH69, twice as much IFN was produced than in cells obtained from nonsensitized mice. This increase was explained by the presence of both sensitized macrophages and T‐cells in a reconstitution experiment.
A Study of Cyanide‐Insensitive Respiration in the Genus Dekkera and Brettanomyces
Authors: Blondin Bruno , Gonde Pierre , Ratomahenina Robert , Arnaud Alain , Galzy Pierre
Published: November 2013

Dekkera intermedia and Brettanomyces custersii were shown to have a respiratory pathway resistant to cyanide, antimycin A, and azide. This respiration remained sensitive to salicylhydroxamic acid (SHAM). The “cyanide‐resistant” respiration was induced mainly at the end of the growth phase and could reach 50% of total respiratory capacity.

The mitochondrial “petite colony” mutation had no effect on this oxidation pathway. The presence of this respiration pathway in these strains constitutes a compensation mechanism for the reducing activity of acetaldehyde dehydrogenase. This alternate pathway would thus be a fundamental element, of the Custer effect, a characteristic feature of these strains.
Diarrhea in Neonatal Piglets Caused by K99+ST+ Enterotoxigenic Escherichia coli
Authors: Shimizu Mikio , Sakano Tetsuya , Yamamoto Junya , Sakurai Ken‐Ichiro
Published: November 2013

Enterotoxigenic Escherichia coli strains were isolated from the feces of 34.4% of 64 diarrheaic neonatal piglets on seven farms. Of a total of 518 isolates, 86 (16.6%) were enterotoxigenic and grouped into four phenotypes: K99+ST+ (K99 pilus antigen and heat‐stable enterotoxin producing, 36 strains), ST+ (37 strains), K88+LT+(K88 pilus antigen and heat‐labile enterotoxin producing, 11 strains), and K88+ST+ (2 strains). K99+ST+ and ST+ isolates showed multiple drug resistance and most of those (58.3% and 56.8%, respectively) belonged to O serogroup 101. A K99+ST+ isolate proved to be capable of inducing diarrhea and death in hysterectomy‐produced colostrum‐deprived piglets when orally inoculated.
Adjuvant Activity of Klebsiella O3 Lipopolysaccharide: Comparative Study Using Defined Uniform Salt Forms
Authors: Kato Nobuo , Kido Nobuo , Ohta Michio , Naito Setsuko , Nakashima Izumi
Published: November 2013

Previously we showed that Klebsiella O3 lipopolysaccharide (KO3 LPS) is much more potent than other kinds of LPS including Escherichia coli O127 LPS (EO127 LPS) in adjuvant activity in augmenting antibody response and delayed‐type hypersensitivity to protein antigens and in the ability to enlarge the regional lymph node. Various denned uniform salt forms, the triethylamine, sodium, potassium, ammonium, tris(hydroxymethyl)ammomethane, and calcium salt forms, of KO3 LPS and EO127 LPS were prepared by removing basic materials present in LPS preparations by electrodialysis and neutralizing the electrodialyzed LPS preparations with various kinds of alkali. The triethylamine salt form showed the best solubility and consisted of the smallest granules and, on the other hand, the calcium salt form showed the lowest solubility, compared with the natural form and the other uniform salt forms. Even if the natural forms of KO3 LPS and EO127 LPS were converted to the defined uniform salt forms, adjuvanticity of KO3 LPS and EO127 LPS in augmenting delayed‐type hypersensitivity to ovalbumin and the ability to enlarge the regional lymph node did not significantly differ from those of the respective natural forms. From these results it is concluded that the difference in strength of the adjuvanticity between KO3 LPS and EO127 LPS is not due to the difference in their salt forms, solubility or physical state. Moreover, there were no significant differences in lethal toxicity for mice by the intraperitoneal route among the natural form and all the uniform salt forms of KO3 LPS tested.
Environmental Mycobacteria in Korea
Authors: Jin Byoung Won , Saito Hajime , Yoshii Zensaku
Published: November 2013

Environmental mycobacteria in Korea have been investigated by examining 54 soil, 111 house dust, 63 well water, and 98 sewage samples collected from 123 randomly selected areas in Korea during the fourth nationwide tuberculosis prevalence survey in 1980.

A variety of mycobacteria were isolated from 76% of soil, 67% of sewage, 43% of well water, and 7% of house dust samples. Some samples yielded more than one species; thus 56 strains were obtained from soil, 107 strains from sewage, 48 strains from well water and 8 strains from house dust. Mycobacterium fortuitum was the most common species of environmental mycobacteria in Korea and the species was distributed equally in all types of samples tested. The M. terrae complex was also one of the common species of environmental mycobacteria and it seemed to be more abundant in water samples than in soil. Scotochromogenic slow growers M. scrofulaceum and M. gordonae were common microbes in soil and water samples, although the latter was more frequently detected in water samples. Scotochromogenic rapid growers M. flavescens and M. phlei, and photochromogenic rapid grower M. vaccae were isolated more frequently from sewage or water samples than from soil. Nonphotochromogenic rapid growers M. chelonei (chelonae) and M. smegmatis were isolated mostly from sewage and the former was rarely found in soil and well water samples. The clinically important species M. avium‐intracellulare complex was found less frequently in all types of test samples.
Immunogenic Dialyzable Factor Derived from a Ribosomal Fraction of Salmonella typhimurium
Authors: Kita Eiji , Yasui Koichi , Yasui Kiyoshi , Matsuda Yasushi , Matsuda Keiji , Kashiba Shuzo
Published: November 2013

Dialyzable factors (DF) were prepared from ribosomal fractions of several organisms including rough mutants of Salmonella typhimurium LT2, salmonella species of different serogroups, other enteric bacteria and gram‐positive organisms, and tested for their immunogenicity against S. typhimurium infection in mice. All of them conferred local resistance on mice challenged intramuscularly with S. typhimurium LT2 in the early stage of immunization before the establishment of delayed‐type hypersensitivity (DTH) to salmonella antigens. Although DFs of enteric bacteria including rough mutants of S. typhimurium induced DTH to salmonella antigens, only DF of a two‐heptose mutant of S. typhimurium LT2 afforded significant mouse protection but others only prolonged the mean time to death. DF of Listeria monocytogenes induced the cross‐reacting immunity which afforded the low level of mouse protection as well as an increase in mean time to death without inducing DTH. Passive transfer of anti‐O antibody did not enhance the mouse protection provided by each DF. Resistance conferred by DF of S. typhimurium LT2 consisted of two phases: (i) nonspecific macrophage activation resulting in reduction of organisms at the infected site, which became active in the early stage of immunization and (ii) salmonella‐specific immunity capable of preventing systemic infection, which became active in the late stage of immunization.
Ability of Various Oral Bacteria to Bind Human Plasma Fibronectin
Authors: Imai Susumu , Okahashi Nobuo , Koga Toshihiko , Nisizawa Tosiki , Hamada Shigeyuki
Published: November 2013

The present study describes the ability of various oral bacteria to bind human plasma fibronectin (PFN). Avid binding of 125I‐PFN was found for Streptococcus mutans (serotypes a to h), Streptococcus sanguis, group A Streptococcus pyogenes and Staphylococcus aureus, while other gram‐positive bacterial species tested demonstrated only weak or negligible PFN binding ability. Two gram‐negative bacterial species, Bacteroides gingivalis and Escherichia coli, did not significantly bind PFN. 125I‐PFN binding to S. mutans 6715 cells was decreased by pretreatment with unlabeled PFN, and the radiolabeled PFN bound to the cell surface was released on addition of unlabeled PFN. Strong inhibition of 125I‐PFN binding to S. mutans 6715 cells was obtained by protease pretreatment, while partial inhibition was also observed following treatment with acid, alkali, lipase, and monoclonal anti‐polyglycerophosphate. These results suggest that PFN binding to S. mutans cells is reversible and that PFN receptors on the cell surface appear to be heat‐stable multiple proteins.
Increased Sister Chromatid Exchange in Human Lymphocyte Cultures Infected with Mycoplasma
Authors: Ito‐Kuwa Shoko , Aoki Shigeji
Published: November 2013

The effects of fermenting, poorly arginine‐utilizing Mycoplasma fermentans and arginine‐utilizing Mycoplasma salivarium on the frequency of sister chromatid exchange (SCE) in cultured human lymphocytes were examined. M. fermentans caused no apparent mitosis inhibition of lymphocytes and the increase in SCE frequency was dependent on the inoculum size of the mycoplasma. An evident increase in SCE frequency was observed in lymphocytes infected with smaller inoculum sizes of M. salivarium whereas there was mitosis inhibition of lymphocytes infected with larger inoculum sizes of the mycoplasma. In lymphocyte cultures infected with M. salivarium, the addition of arginine to the culture medium reduced mitosis inhibition but did not diminish the increase in SCE frequency, indicating that arginine depletion was not involved in causing the induction of SCEs in mycoplasma‐infected lymphocytes. With regard to the genetic effectiveness of SCE, these results suggested that mycoplasmas are capable of inducing cytogenetic changes in infected host cells.
Protecting Effect of Chitin and Chitosan on Experimentally Induced Murine Candidiasis
Authors: Suzuki Ko , Okawa Yoshio , Hashimoto Kazuhiko , Suzuki Shigeo , Suzuki Masuko
Published: November 2013

Chitin and chitosan were found to exhibit a protective effect on mice administered these polysaccharides intraperitoneally against infection of the viable cells of Candida albicans NIH A‐207 strain. A significant difference was observed between the protective effects of chitin and chitosan, i.e., chitin was much more effective than chitosan when the C. albicans cells were challenged via the intravenous route. In intraperitoneal inoculations of C. albicans cells, however, chitosan provided stronger resistance for mice than chitin.

It has also been revealed that the number of polymorphonuclear leukocytes from circulating blood of chitin‐administered mice increased remarkably compared with that of untreated and chitosan‐treated mice, and that the increase of active oxygen‐generating phagocytic cells was significant. On the other hand, the number of peritoneal exudate cells (PEC) and the amounts of active oxygen generated from these cells in chitosan‐treated mice were larger than those of chitin‐treated mice. However, candidacidal activities of PEC per fixed cell number in mice treated with chitin or with chitosan were almost the same and greater than those of untreated mice.
Biological Activities of Partially Purified Elastase Produced by Flavobacterium meningosepticum
Authors: Miyazaki Shuichi
Published: November 2013

Injection of viable cells of Flavobacterium meningosepticum at doses of 106 colony‐forming units/eye caused ophthalmia in rabbits' corneas.

Elastase, free of protease activity, was partially purified from broth cultures of F. meningosepticum ATCC12535 and TMS516. The optimum pH of the elastase activity was about 8.0. The activity was suppressed by Ni, Zn, Co, EDTA, and o‐phenanthroline, and accelerated by Cu and Fe.

The minimum dose of the elastase causing ophthalmia in the eyes of rabbits was 5 μg/eye (0.065 elastase units). The minimum lethal dose of the elastase for suckling mice by intracerebral injection was 0.4 μg (0.0052 elastase units)/suckling mouse, and its minimum lethal dose for adult mice by intraperitoneal injection was 740 μg/mouse (9.52 elastase units).
Further Studies of the Use of Cycloheximide for Cultivating Mycobacterium lapraemurium in Cell Culture
Authors: Matsuo Yoshiyasu
Published: November 2013

The advantage of using cycloheximide for cultivating Mycobacterium lepraemurium in cell culture was further demonstrated. Continuous multiplication of the bacillus in successive subcultures was obtained in MFP, HEp‐2 and Vero cells maintained in culture medium containing 0.1 μg of cycloheximide per ml. Growth characteristics were comparable to those observed in the cultures of A31 cells previously reported. The procedure was simple and convenient. Comparable results, however, have not been obtained in cultures of other established cell lines, HeLa 229, L, MDCK, and Neuro‐2a.
Dengue Type 2 Virus Infection in Human Peripheral Blood Monocyte Cultures
Authors: Hotta Hak , Wiharta Agus S. , Hotta Susumu , Homma Morio
Published: November 2013

Dengue type 2 virus (D2V) infection in cultured human monocytes was studied. D2V permissiveness of the monocytes was enhanced when the cells were inoculated with D2V in the presence of either polyclonal or type‐specific monoclonal anti‐dengue antibody. The enhancement of D2V permissiveness mediated by the antibodies was more clearly demonstrated when the monocytes had been treated with trypsin before virus inoculation, though treatment of the cells with trypsin alone decreased D2V permissiveness. The enhancement of infection by type‐specific neutralizing monoclonal antibody suggests that the D2V particles possess at least two antigenic determinants closely associated with virus infectivity.

Infectious center assays revealed that the infection enhancement in the presence of the antibodies was due primarily to an increase in the number of D2V‐infected cells, and that only a small proportion of the monocyte population supported D2V replication. The virus‐permissive monocytes did not bear HLA‐DR antigens on their cell surface. The presence of nonadherent lymphocytes in the monocyte cultures before D2V inoculation did not affect the D2V permissiveness of the monocytes.

Treatment of cultured monocytes with the synthetic adjuvants N‐acetylmuramyl‐L‐alanyl‐D‐isoglutamine (MDP) and its lipophilic derivative, [B30]‐MDP, did not significantly affect the D2V permissiveness of the cells.
Determination of Hepatitis B Virus DNA in Serum by Molecular Hybridization
Authors: Shimizu Yoshitaka , Ida Setsuko , Matsukura Toshihiko , Yuasa Tazuko
Published: November 2013

Hepatitis B virus (HBV) DNA was detected by direct spotting of alkali‐denatured serum on a nitrocellulose filter and molecular hybridization with cloned HBV DNA as the probe. Measurement of the autoradiographic signals as the intensity of hybridization allowed the quantitation of HBV DNA content in serum specimens in reference to cloned HBV DNA. Direct spotting of denatured serum was approximately three times as sensitive as the conventional method in which proteinase‐treated serum was extracted with phenol‐chloroform. The intensity of hybridization with 25 specimens of HB virion concentrates correlated well with DNA polymerase activity (r = 0.89, P < 0.01).
Cross Wall Synthesis and the Arrangement of the Wall Polymers in the Cell Wall of Staphylococcus spp.
Authors: Amako Kazunobu , Umeda Akiko
Published: November 2013

The growing process and the fine structure of the cross wall of Staphylococcus were investigated by electron microscopy. Examination of the tangentially sectioned cross wall revealed that it was initially synthesized as a thin cell wall layer by an invaginated cytoplasmic membrane. The wall thickness soon increased by additional synthesis of the wall from the cytoplasmic membrane located at the side region of the cross wall. Scanning electron microscopic observation of sodium dodecyl sulfate‐treated and mechanically separated cross walls revealed that the outer surface of the cross wall exhibits regular circular structures and the inner surface showed has an irregular surface. This indicates that cell wall materials were arranged in a regular circular manner in the initially synthesized thin layer. It is conceivable that in Staphylococcus spp. two cell wall synthesizing systems are present: wall‐elongation synthesis in which wall materials are arranged in a regular circular manner and wall‐thickening synthesis in which wall materials are arranged in an irregular manner.
Enhancement of Endotoxicity and Reactivity with Carbocyanine Dye by Sonication of Lipopolysaccharide
Authors: Ogawa Yoshiyuki , Kanoh Seizaburo
Published: November 2013

The specificity of endotoxin (lipopolysaccharide, LPS) in the carbocyanine dye reaction was investigated, and then a stoichiometric study of the dye‐LPS interaction was conducted with attention to the relationship of biological activities of LPS to the reactivity with the dye. Absorption maxima of some bacterial components in the dye reaction were as follows; LPS from both Escherichia coli and Pseudomonas aeruginosa and lipid A from E. coli LPS, 465 nm; Shigella flexneri LPS, 460 nm; Salmonella minnesota R595 glycolipid, 470 nm; polysaccharide from E. coli LPS, 650 nm; yeast RNA, 620 nm; streptococcal M protein and pyrogenic exotoxin, 610 nm; and free fatty acids, 445–450 nm. The absorbance at 465 nm was increased approximately threefold by sonicating LPS for 1–3 min, which roughly paralleled the decrease in turbidity of the LPS aqueous solution. The Limulus amoebocyte lysate (LAL) gelation activity of LPS increased 10‐fold when LPS was sonicated for 0.5–5 min, but it decreased to the control level after further treatment. This decrease, however, was overcome by sonication in the presence of 5 mmol of L‐ascorbic acid used as an antioxidant. The LAL gelation activity of LPS was inactivated in parallel with an increase in the ratio (w/w) of dye to LPS from 1.73 to 6.90 in the dye‐LPS mixture. Pyrogenicity of LPS was also clearly inactivated when the ratio was over 1.73. The ratios of the height of the β band at 465 nm (dye‐LPS complex) to that of the α band at 510 nm (free dye) were increased by sonicating LPS, indicating that the binding character, or stacking tendency, was increased by sonicating LPS.
Colonial Morphology of Treponemes Observed by Electron Microscopy
Authors: Umemoto Toshihiko , Namikawa Isamu , Yamamoto Mituhito
Published: November 2013

The colonial morphology of three strains of cultivable, nonpathogenic treponemes including a human oral treponeme was examined by light and electron microscopy. Treponema phagedenis strains Kazan and Reiter produced large white colonies on the surface of solid media composed of sterility test broth, 0.9 to 3.1% agar, rifampin, and 12.5% rabbit or horse serum. A human oral treponeme, strain G7201, grew as diffused white zones on 0.9 to 3.1% agar plates. Under the cultural conditions employed agar concentrations slightly affected the time of appearance of colonies of the three strains of treponemes.

When the colonies of these three strains were viewed by scanning electron microscopy, differences in their colonial morphology were observed. The 11‐day‐old colonies of human oral strain G7201 were very small, 5 to 15 μm in diameter, and had a slight irregular border. Kazan treponemes developed circular, entire and low convex colonies. Scanning and transmission electron microscopy revealed that the colonies of Reiter treponemes contained spherical forms almost up to 5 μm in diameter, each consisting of an outer membrane and a treponemal main body. They were very similar to the spherical bodies produced by strain G7201 in sucrose‐containing broth.
Role of Serum Factors in the Phagocytosis of Weakly or Heavily Encapsulated Cryptococcus neoformans Strains by Guinea Pig Peripheral Blood Leukocytes
Authors: Ikeda Reiko , Shinoda Takako , Kagaya Keiko , Fukazawa Yoshimura
Published: November 2013

We investigated the opsonic activity of the serum factors affecting phagocytosis of Cryptococcus neoformans in vitro to elucidate the role of humoral factors in the host defense mechanisms against cryptococcosis. Two strains of C. neoformans, one heavily and one weakly encapsulated, were used. Guinea pig peripheral blood leukocytes (PBLs) were used for phagocytosis. The viable weakly encapsulated cells were ingested effectively by PBLs, in the presence of guinea pig normal fresh serum, while the heavily encapsulated cells were not ingested. Neither immune serum, its IgG fraction alone, nor heated serum promoted the phagocytosis of either the weakly or heavily encapsulated strain. On the other hand, immune serum promoted adherence of PBLs to viable cells of the heavily encapsulated strain, forming rosettes in the presence of fresh serum. A substantial amount of C3b component was detected on yeast cells when weakly encapsulated cells were incubated with human fresh serum, or heavily encapsulated cells were incubated with rabbit immune serum together with human fresh serum. Serum chelation experiments also indicated that the factors involved in the alternative complement pathway are opsonins for the weakly encapsulated strain. These results suggest that the alternative pathway plays an important normal opsonic role for weakly encapsulated strains and that specific antibody plays an immune opsonic role for heavily encapsulated strains of C. neoformans via the classical pathway of complement activation.
Demonstration of T Antigens on the Surface of Cells Transformed with Primate Polyoma Viruses
Authors: Ohno Tadayoshi , Yamaguchi Nobuo , Uchida Seijiro
Published: November 2013

Primate polyoma virus‐transformed hamster, mouse, and rat cell lines were examined by indirect immunofluorescence staining for cell surface‐associated T antigens, by using a rabbit antiserum prepared against sodium dodecyl sulfate‐denatured large T antigen of simian virus 40 (anti‐SV40‐SDS‐T serum). Positive surface staining was shown not only on SV40‐transformed cells, but also on BK and JC virus‐transformed cells. In contrast, normal cells and cells transformed with mouse polyoma‐, human adeno‐, and murine sarcoma viruses were negative. The data on SV40‐transformed cells confirmed the reports of others demonstrating the cell surface location of SV40 large T antigen, and the data on BK and JC virus‐transformed cells proved that these cells have cell‐surface T antigens that cross‐react with anti‐SV40‐SDS‐T serum.
Germinability and Heat Resistance of Spores of Clostridium difficile Strains
Authors: Nakamura Shinichi , Yamakawa Kiyotaka , Izumi Junko , Nakashio Satoshi , Nishida Shoki
Published: November 2013

Out of 111 Clostridium difficile strains, 108 produced spores in numbers of more than 105/ml and the remaining three did not produce any spores in brain heart infusion medium. The germination frequency in the medium without lysozyme varied widely from strain to strain, ranging from less than 10−8 to 100, and in 77 of the 108 strains the germination frequency was 10−5 or less. The spores, when treated with sodium thioglycollate and then inoculated into the medium containing lysozyme, germinated in all of the 108 strains at a frequency of 10−0.5 or more. The spores of two strains germinated at a frequency of more than 10−0.5 in all methods. Spores of C. difficile strains were fairly highly heat‐resistant; D100C values ranged from 2.5 to 33.5 min.
Inhibition of Cortex Hydrolysis during Spore Germination by CdCl2
Authors: Nakatani Yoshihiro , Imagawa Masayoshi , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

When the spores of Bacillus megaterium QM B1551 (ATCC 12872) were incubated with 5 mM CdCl2 at 30 C, they underwent the early germination events, such as loss of heat resistance and release of calcium dipicolinate, in the same way as when they were germinated by glucose+KNO3. However, germination by CdCl2 caused no increase in the reducing groups in the cortex and no excretion of glucosamine‐containing materials due to the hydrolysis of the cortex peptidoglycan. Addition of CdCl2 at any time during germination by glucose + KNO3 inhibited the release of glucosamine‐containing materials from the spores, whereas removal of cadmium from the CdCl2‐germinated spores by treatment with cysteine restored the hydrolysis of peptidoglycan.

These results suggested that CdCl2 caused the early events of spore germination but prevented the spores from undergoing the events following germination by inhibiting the enzymatic lysis of the cortex peptidoglycan. The conclusion from the study is that cortex degradation is not always required for the initiation of germination.
Lateral Flagellar Antigen of Vibrio alginolyticus and Vibrio harveyi: Existence of Serovars Common to the Two Species
Authors: Shinoda Sumio , Nakahara Noriko , Uchida Emiko , Hiraga Michie
Published: November 2013

The antigenicity of lateral (L‐) flagella of two marine vibrios, Vibrio alginolyticus and V. harveyi, was studied, and the two species were found to have common antigenicity of their flagella. Antisera against L‐flagella were prepared by immunizing rabbits with highly purified L‐flagellar filaments. H‐Agglutination tests with the anti‐L‐flagella antisera showed that four H‐serovars existed in these species and that two of them were shared by the two species. Cross reactivity between H‐serovars of these two species and other vibrios having lateral flagella, such as V. parahaemolyticus, V. campbellii, V. proteus, or V. fluvialis, was not observed in the H‐agglutination test, although partial common antigenicity was observed in the gel diffusion test with flagellin monomers. These observations suggest that surface antigenic determinants of the lateral flagella of V. alginolyticus and V. harveyi are specific to these two species but internal antigenic determinants buried in the flagellar filaments are partially shared with other vibrio species.
Production of Monoclonal Antibodies against Legionella pneumophila Serogroup 1
Authors: Miyazaki Takashige , Koga Hironobu , Nakashima Manabu , Tomonaga Akimitsu , Kohno Shigeru , Hirota Masaki , Saito Atsushi , Hara Kohei , Watanabe Takeshi
Published: November 2013

Four monoclonal antibodies to Legionella pneumophila Philadelphia 1 were produced by the fusion of immunized BALB/c lymphocytes to a murine myeloma cell line. Two (Lpl‐1 and Lpl‐3) of the four monoclonal antibodies reacted with 14 L. pneumophila serogroup 1 strains, and the other (Lpl‐2 and Lpl‐4) reacted with only three out of 20 strains tested. These four monoclonal antibodies did not bind to any strains of L. pneumophila serogroup 2–7 and other Legionella species. In addition, it has been shown that these monoclonal antibodies may be useful not only for subserotyping of L. pneumophila but also for retrospective diagnosis using immunopathological methods.
Fatty Acid Composition and Shwartzman Activity of Lipopolysaccharides from Oral Bacteria
Authors: Mashimo Jun‐Ichi , Yoshida Michiko , Ikeughi Kuniko , Hata Seiichi , Arata Satoru , Kasai Nobuhiko , Okuda Katsuji , Takazoe Ichiro
Published: November 2013

The composition and the nature of the linkage of fatty acids and the Shwartzman activity of lipopolysaccharide (LPS) preparations derived from oral gram‐negative bacteria including Bacteroides gingivalis, Bacteroides loesheii, Eikenella corrodens, Fusobacterium nucleatum, and Actinobacillus actinomycetemcomitans were examined. 3‐Hydroxylated and nonhydroxy fatty acids of various chain lengths were found in all of the LPS preparations. All nonhydroxy fatty acids were found to be ester‐bound, and part of the 3‐hydroxy fatty acids in the LPS of B. gingivalis, E. corrodens, F. nucleatum, and A. actinomycetemcomitans were shown to be involved in ester linkage. It was also suggested that the hydroxy group of the ester‐bound 3‐hydroxy fatty acid of the LPS of F. nucleatum and A, actinomycetemcomitans is at least partly substituted by another fatty acid, but in the LPS of B. gingivalis and E. corrodens it is not. The main amide‐linked fatty acid of the LPS of B. gingivalis, E. corrodens, F. nucleatum, and A. actinomycetemcomitans was 3‐hydroxyheptadecanoic, 3‐hydroxydodecanoic, 3‐hydroxyhexadecanoic, and 3‐hydroxytetradecanoic acid, respectively. The results of the Shwartzman assay showed that the E. corrodens LPS was the most active among the preparations tested, and that the Shwartzman toxicity of Bacteroides LPS is extremely low.
Gene Structures of Low‐Neurovirulent Vaccinia Virus LC16m0, LC16m8, and Their Lister Original (LO) Strains
Authors: Sugimoto Masanobu , Yasuda Atsushi , Miki Keizaburo , Morita Michio , Suzuki Kazuyoshi , Uchida Nobuyuki , Hashizume Soh
Published: November 2013

Vaccinia viruses LC16m0 and LC16m8 are temperature‐sensitive and low‐neurovirulent variants derived from the Lister (Elstree) (LO) strain. Analyses of genome DNAs by digestion with restriction endonucleases and cross‐hybridization of the digested fragments revealed that LC16m0 and LC16m8 possess a new XhoI site in addition to the 14 XhoI sites of LO. This new site is located at about 12 × 106 daltons from the right terminal end. There was no significant difference in the genome structures between the LC16 variants and LO except the new XhoI site and their terminal fragments which were not identified in LO owing to their heterogeneity. With HindIII digested fragments, there was no difference among the three viruses. This complete mapping raised the possibility that the putative gene responsible for temperature sensitivity and neurovirulence is located at the region of the XhoI site found in LC16m0 and LC16m8.
Actinomyces viscosus Cell‐Free Synthesis of Extracellular Slime Polysaccharide
Authors: Ooshima Takashi , Kuramitsu Howard K.
Published: November 2013

A cell‐free extract of Actinomyces viscosus T14Av catalyzed the synthesis of extracellular N‐acetylglucosamine‐rich slime polysaccharide. The activity was localized in the cytoplasmic membrane fraction and required the presence of ADP‐glucose and UDP‐N‐acetylglucosamine. Maximal activity was demonstrated at pH 7.5 and also required the presence of divalent cations such as Mg2+ or Mn2+. Extracellular slime appeared to serve as a primer for slime biosynthesis. The antibiotic tunicamycin acted as an inhibitor of slime formation. However, another glucosamine analogue, amphomycin, as well as the antibiotic bacitracin produced moderate stimulatory effects on slime biosynthesis.