bluuebunny/crossref_metadata_2025 huggingface dataset:
Acid and Alkaline Phosphatases of Vibrio parahaemolyticus
Authors: Sakaguchi Osamu , Yokota Katushi , Koshi Tomoyuki
Published: November 2013

Characteristics of phosphatases of Vibrio parahaemolyticus A55 K+ 105 were investigated. Phosphatases were extracted from intact cells in a 3% NaCl solution by gentle treatment on a reciprocal shaker for 15 min. The pH dependence of phosphatase of the extracts showed two optima; one optimum was found at pH 5.1–5.7, the other at pH 10.5–11.1, and the results were identical with that of resting cells. The activities of both acid and alkaline phosphatases were maximum in the presence of 0.1 m salt (KCl or NaCl). The production of alkaline phosphatase of resting cells in the induction medium containing p‐nitrophenylphosphate was largely suppressed by the addition of 0.5% phosphate into the medium. The alkaline phosphatase of the extracts was stimulated about 2.5‐fold by the addition of 10−3m Mg++ or Zn++, whereas it was inhibited completely by 10−3 m ethylenediaminetetraacetic acid. On the other hand, Co++ activated acid phosphatase to 98‐fold. Both acid and alkaline phosphatases were found in the culture filtrated after cultivation of the organism in a 3% NaCl‐nutrient broth medium for 16 hr.
Studies on Temperate Phages of Lactobacillus salivarius
Authors: Tohyama Kiyoshi , Sakurai Toshizo , Arai Hiroshi , Oda Akira
Published: November 2013

Twenty‐three temperate phages of Lactobacillus salivarius isolated from human feces were studied as to their morphological, biological, and serological properties. (1) Among 30 strains of L. salivarius tested, 23 strains were lysed by induction with mitomycin C (MC). In all these lysates, phage particles were detected by electron microscopic examination. (2) These phages were morphologically divided into three groups: particles with a regular hexagonal head and a long flexible tail; particles having a regular hexagonal head with or without a short tail‐like structure; particles with an elongated head and a long noncontractile tail. (3) Only two, phage 223 having an elongated head and phage 227 with a regular hexagonal head and a long noncontractile tail, produced tiny and very turbid plaques on several host bacteria. Six phages could produce only inhibition zones, ranging from complete inhibition through partial inhibition to normal growth by a serial dilution spot test. (4) All these killer particles could also inhibit the growth of their producer cells. (5) A serological relationship was observed between temperate phages and killer particles, and this was somewhat consistent with the morphological groupings.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Makidono Reiko , Takeya Kenji
Published: November 2013

Delayed hypersensitivity against hamster erythrocyte antigen was examined after sensitization with hamster erythrocytes (HRBC) in Freund's complete adjuvant (FCA). Extent of the delayed hypersensitivity was determined by the migration inhibition test, the peritoneal macrophage disappearance test and the skin test in the ear using solubilized HRBC as the test antigen. 1) Delayed hypersensitivity against HRBC developed earlier in high‐responder SL mice than in low‐responder C57BL/6 mice after sensitization. The period required for development of the delayed hypersensitivity in AKR mice was intermediate between periods in high‐responder SL mice and low‐responder C57BL/6 mice. 2) After sensitization with HRBC in FCA, a delayed hypersensitive state without detectable antibody production persisted until day 12 in high‐responder SL mice and until day 16 or later in low‐responder C57BL/6 mice. 3) Delayed hypersensitivity against HRBC antigen persisted even after the appearance of circulating antibody which occurred late after sensitization with HRBC in FCA or after intravenous injection of HRBC into sensitized mice.
Formation of Virus‐Inhibiting Factor or Interferon in Splenectomized Mice
Authors: Maehara Nobutoshi , Nagano Yasuiti
Published: November 2013

Splenectomized mice were injected intravenously with Escherichia coli endotoxin or Newcastle disease virus at different intervals. Serum and various organs were taken at different intervals and tested for virus‐inhibiting factor (IF) or interferon titer. Production of endotoxin‐induced IF was depressed immediately after splenectomy. This depression was recovered in six weeks. IF was then formed mainly in the lung and liver. Splenectomy did not immediately affect IF production induced with virus. In the course of time, the production of late‐appearing virus‐induced IF was gradually depressed, whereas the formation of early‐appearing virus‐induced IF was progressively enhanced. These results were interpreted as follows: (1) Endotoxin‐induced IF and virus‐induced early IF are formed by different mechanisms, although these IFs have similar physicochemical properties; (2) In splenectomized mice, the activity of spleen to form endotoxin‐induced IF may be compensated by certain cells in various organs, whereas that for production of virus‐induced late IF is not; (3) The production of virus‐induced late IF depends on the spleen by a somewhat indirect way. When mice were inoculated with virus within 60 days after splenectomy, the serum IF titer was higher than those of organs. These findings suggested that the organs examined were not the major sources of virus‐induced serum IF in splenectomized mice.
Staphylococcal Enterotoxin D
Authors: Igarashi Hideo
Published: November 2013

Purification of enterotoxin D from Staphylococcus aureus 494 was attempted by utilizing the following techniques: Concentration of bacterial culture supernatant with polyethylene glycol 20000, CM‐cellulose column chromatography, Sephadex G‐100 gel‐filtration, chromatography on a DEAE cellulose column with concave gradient elution and gel‐filtration with Sephadex G‐50. The final preparation obtained gave a single precipitin line with anti‐crude enterotoxin D serum and no precipitation with anti‐enterotoxin A, B or C when it was tested by Ouchterlony's technique. It was capable of producing 100% emesis in monkeys at about 1.25 μg of protein per kg body weight. Antiserum prepared by injecting rabbits with the purified preparation gave a single precipitin line with concentrated crude toxin, and it also gave a single arc when tested in immunoelectrophoresis with concentrated crude toxin. A type‐specific neutralization effect in monkeys was obtained. Thus, enterotoxin D has been identified with a purified preparation.
Biochemical Properties of a Cephalosporin β‐Lactamase from Pseudomonas aeruginosa
Authors: Yaginuma Satoshi , Sawai Tetsuo , Ono Hideo , Yamagishi Saburo , Mitsuhashi Susumu
Published: November 2013

The cephalosporin β‐lactamase from Pseudomonas aeruginosa GN918 was purified using CM‐Sepha‐dex column chromatography. The resulting preparation gave a single protein peak on electrofocusing column chromatography and a single protein band on polyacrylamide‐gel electrophoresis. The specific enzyme activity was 22 950 units per mg of the purified enzyme protein. The optimal pH was 7.5 and the optimal temperature was 40 C for the hydrolysis of cephaloridine. Isoelectric point was 8.7 and the approximate molecular weight of the enzyme was found to be 34 000±2000. The enzyme activity was inhibited by iodine, p‐chloromercuribenzoate and semi‐synthetic penicillins. The enzymological properties of the isolated preparation have been compared with β‐lactamases derived from other gram‐negative enteric bacteria.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Kuroiwa Ataru , Mashiba Harukazu , Takeya Kenji
Published: November 2013

1) A subcutaneous injection of hamster erythrocytes (HRBC) in Freund's complete adjuvant (FCA) or an intravenous injection of hamster lymph node (HLN) cells suppressed antibody production against HRBC in the low‐responder C57BL/6 and AKR mice, when HRBC in saline were given on the same day; 2) The suppressing effect of such treatments was neither detectable in the high‐responder SL mice, nor in the C57BL/6 mice, which had been pre‐sensitized with HRBC in FCA or hamster lymphoma cells; 3) Positive reactions of the peritoneal macrophage disappearance test and the enhanced antibody production were detected seven days after treatment with HRBC in FCA and HRBC in saline, or HLN cells and HRBC in saline; 4) The suppressing effect of such simultaneous treatments on anti‐HRBC antibody production was eliminated by a transfer of normal syngeneic thymus cells to AKR mice or a transfer of thymus cells from SL to C57BL/6 mice. Suppression of the antibody production in the low‐responder mice by the described simultaneous treatments may be due to a competitive involvement of HRBC‐specific thymus‐derived cells (T cells) in the developmental stages of delayed hypersensitivity and antibody production. High‐responder SL mice appear to have enough T cells for development of the delayed hypersensitivity and as helper cells in antibody production. These results appear to support the concept that T cells for delayed hypersensitivity and antibody production to HRBC antigen are derived from the same original pool.
Studies on Temperate Phages of Lactobacillus salivarius
Authors: Tohyama Kiyoshi
Published: November 2013

Comparative studies were carried out on the mode of action of the defective temperate phage 208 against the homologous lysogenic strain S‐208 and a nonlysogenic strain S‐161 of Lactobacillus salivarius. Treatment of both strains with phage 208 led to a specific inhibition of protein synthesis, cell killing without any reversion of protein synthesis, and of viable counts by treatment with trypsin. Killing action of phage 208 followed a single hit kinetics for nonlysogenic S‐161 and S‐208 No. 006, which is a cured strain of S‐208, whereas, two to five hit kinetics was obtained for lysogenic S‐208. Phage particles exposed to ultraviolet light (5.7 kergs/cm2) also killed S‐161 with a single hit kinetics and S‐208 with a 3‐hit kinetics. However, the kinetics of killing approached a single hit when the protein synthesis of S‐208 was inhibited by chloramphenicol prior to the phage addition. Based upon these results, the possible mechanisms of immunity breakdown and of subsequent cell killing were discussed.
Antibody Responses in Germ‐Free Chickens to Bacteria Isolated from Various Sources
Authors: Morishita Yoshiyuki , Mitsuoka Tomotari
Published: November 2013

The germ‐free chickens were mono‐ or dicontaminated with various species of facultative anaerobes and obligate anaerobes isolated from chickens, humans, pigs, and fermentation products, and their sera were studied for agglutinins against these organisms. After contamination, gnotobiotic chickens developed relatively high titers of agglutinins against Streptococcus faecalis var. liquefaciens from a chicken and obligate anaerobes such as Bifidobacterium thermophilum (strain T 111), Bacteroides sp. (Au 21–27), and Fusobacterium (Sphaerophorus) sp. (EBF 59/96p) from chickens, Bacteroides sp. (A 269–5) from a human, and B. thermophilum (P 2–91) and Fusobacterium (Sphaerophorus) sp. (PNC 2–23) from pigs. On the contrary, little or no detectable agglutinins were observed against Bacteroides sp. (Ch 220–17) from a chicken, Bifidobacterium adolescentis and Bifidobacterium bifidum from humans and Lactobacillus casei and Lactobacillus plantarum from fermentation products. Anti‐Escherichia coli agglutinins were sensitive to 2‐mercaptoethanol, but anti‐B. thermophilum agglutinins were not.
Physicochemical Properties of Bovine Respiratory Syncytial Virus
Authors: Inaba Yuji , Tanaka Yoshio , Omori Tuneyoshi , Matumoto Minoru
Published: November 2013

One of virus strains, isolated from Japanese cattle and serologically identified as bovine respiratory syncytial (RS) virus, was examined. The virus was shown to be an RNA virus with a buoyant density of 1.225, filtrable through a Sartorius membrane filter of 200 nm pore size but not through a 100 or 50 nm filter, sensitive to ether, chloroform and deoxycholate, readily inactivated by trypsin, labile at 56 C and 37 C but stable at −80 C, rapidly inactivated at pH 3.0, and resistant to freeze‐thawing. These properties are compatible with those previously reported for RS virus of human and bovine origin.
Alternate Changes of Surface Antigen(s) in Adenovirus Type 12‐Transformed and Tumor Cells1
Authors: Nakajima Setsuko , Hamada Chuya , Uetake Hisao
Published: November 2013

Alternate changes of specific surface antigen(s) (S antigen) were examined in transformed and tumor cells induced by human adenovirus type 12. All of the hamster and mouse cells transformed in vitro showed ring‐form membrane fluorescence staining by anti‐S antigen rabbit sera, whereas tumor cells, either induced by the virus in vivo or produced by inoculation with the S(+)‐transformed cells, did not show any fluorescence. When the S(–) tumor cells were serially subcultured in vitro, all of them converted to S(+) cells, although more than ten subcultures were necessary. For the S(+) cells to form tumors in hamsters about ten times as many cells were necessary as the S(–) cells. This difference became greater when tumor formation was tested in preimmunized hamsters, while little, if any, when tested in X‐irradiated hamsters. In addition, immunogenicity of the S(+) cells was suggested to be higher than that of the S(–) cells. These findings indicate that the S(+) cells are more immunosensitive and immunogenic than the S(–) cells, and that in vivo conversion from S(+) to S(–) may be due to selection of S(–)‐mutant cells. In vitro conversion from S(–) to S(+) was also suggested to be due to the appearance of S(+)‐mutant cells.
Transmission of Bacteriocinogenicity by Conjugation in Group D Streptococci
Authors: Tomura Tsuneko , Hirano Toyo , Ito Takako , Yoshioka Morimasa
Published: November 2013

Seventy‐seven out of eighty‐one group D streptococcal strains isolated from humans and animals were found to produce bacteriocins that were active on other streptococcal strains of gorup A and D, but inactive on their own cells. On the bases of the spectra of indicator strains, and the sensitivities to heat, chloroform, and trypsin, seven types of bacteriocins were classified. Streptococcus faecalis var. liquefaciens strain 4532 or strain A (liq‐A) was UV‐irradiated, and mutants which lost bacteriocin‐ as well as the β‐hemolysin‐forming activities (Bact–.Hem–) were obtained. Cells of the type I bacteriocin producer (SMr.TCr.Bact‐I+.Hem+) and nonproducer 2025 (PCr.Bact‐I–.Hem–), both belonging to S. faecalis var. liquefaciens, were mixed and incubated in broth. Recombinants (PCr.‐SMs.TCs.Bact‐I+.Hem+) were obtained at a high frequency (5.8% preinoculum size of PCr.‐Bact‐I–.Hem–), and the character was stable for at least ten transfers. In the mixed culture, a marked decrease in the recipient 2025 cell number was observed. The occurrence of recombinants was not inhibited by deoxyribonuclease. A cell‐free filtrate of Bact+.Hem+ cells mixed with Bact–.Hem– cells did pot cause a mutation of the latter combined characters. The transfer of a genetic marker is discussed as an event of the cell‐to‐cell contact.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Nakashima Izumi , Kato Nobuo
Published: November 2013

A study was made to characterize the active substance for the extraordinarily strong adjuvant effect of the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) type 1 Kasuya strain. CPS‐K was fractionated into acidic and neutral CPS‐K by the addition of cetyl‐pyridinium chloride. Neutral CPS‐K exhibited an extremely strong adjuvant effect. The active substance in neutral CPS‐K was precipitable when mixed with a rabbit homologous antiserum. The neutral CPS‐K antigen was serologically distinct from the O antigen and from the acidic CPS‐K which was the type‐specific capsular antigen. Among preparations of neutral CPS‐K from eight different strains of K. pneumoniae tested, the preparation from only one strain (MH‐2) exhibited a strong adjuvant effect comparable to that of the neutral CPS‐K from the Kasuya strain. The neutral CPS‐Ks from Kasuya and MH‐2 strains were antigenically identical. This antigen was not found in all preparations of neutral CPS‐Ks obtained from seven different strains. Preparations of acidic CPS‐Ks from all strains of K. pneumoniae tested with various serologic types including Kasuya and MH‐2 strains were found to exhibit only weak adjuvant effects. The active substance (neutral CPS‐K antigen from Kasuya strain) was shown to form a single peak upon analyses by gel filtration (Sephadex G‐100) and ultracentrifugation. Sedimentation coefficient of the substance was approximately 20 S at a concentration of 5 mg per ml in 0.1 M NaCl. The active substance finally purified by gel filtration contained 65% sugars (as glucose equivalents), 6.8% hexuronic acids, 2.6% hexosamine, 2.3% proteins, and very small amounts of lipids.
Intestinal Resistance in the Experimental Enteric Infection of Mice with a Mouse Adenovirus
Authors: Hashimoto Kazuo , Okada Yasuo , Tajiri Takashi , Amano Hiroshi , Aoki Nobuhiro , Sasaki Shogo
Published: November 2013

The influence of cyclophosphamide (Cy) on the establishment and duration of the intestinal resistance against enteric infection with a mouse adenovirus, strain K87, was examined in inbred mice, strain DK1. When Cy (40 mg/kg/day) was administered to mice for 17 days from the time of virus challenge, a clear prolongation of viral growth and a delayed appearance of neutralizing (NT) antibody in the intestinal wall as well as in the serum were observed. When Cy (40 mg/kg/day, for 14 days) was administered after cessation of viral growth (4 to 6 weeks after virus challenge) and part of the mice were rechallenged with the virus, titers of NT antibody and immunoglobulins became significantly lower than those in control mice not treated with Cy, and regrowth of the virus was observed in eight out of twenty‐five Cy‐treated mice, regardless of the presence or absence of re‐challenge. In this experiment, antibody titers in the intestinal contents of eight virus‐positive mice were significantly lower than those of the remaining seventeen virus‐negative mice. The time when the decrease of intestinal NT antibody was maximum coincided with the time of the maximal frequency of viral regrowth. It was discussed that these facts might present an evidence to support the idea that the intestinal resistance was acquired through local NT antibody belonging to IgA in the intestinal tract.
Antiinfluenzal Effect of Rugulosin
Authors: Nakamura Shinichi , Nii Fusako , Inoue Satoru , Nakanishi Isao , Shimizu Masanao
Published: November 2013

(–)Rugulosin showed an antiinfluenzal effect in the allantoic cavity of chicken eggs as well as in cultured chorioallantois, proving itself to be more potent than 1‐adamantanamine. Also, (–)rugulosin, when administered by inhalation, was as effective for protection of mice against aerosol influenzal infection as orally administered 1‐adamantanamine. The effect of (–)rugulosin seemed to be mainly due to direct inactivation of influenza virus.
Production and Purification of Human Leukocyte Interferon
Authors: Matsuo Akio , Hayashi Saburo , Kishida Tsunataro
Published: November 2013

Interferon production in human leukocyte suspensions induced by Newcastle disease virus is significantly affected by the pH of the culture. The optimal pH for interferon production is higher than the physiological pH level. Human leukocyte interferon has at least three types of surface charge as demonstrated by DEAE‐cellulose column chromatography. The molecular wight of these three types of interferon, as measured by gel chromatography on several kinds of Sephadex is equally 25 000. The main fraction of human leukocyte interferon was purified approximately 1400‐fold by a sequence of procedures which involved ammonium sulfate salting out, CM‐ and DEAE‐cellulose column chromatography and gel filtration on Sephadex G‐50. Human leukocyte interferon of a high specific activity of about 2 130 000 units/mg of protein was thus obtained.
The Role of Macrophages in Candida albicans Infection In Vitro
Authors: Ozato Keiko , Uesaka Ichiro
Published: November 2013

Mouse peritoneal macrophages cultivated in vitro were infected with Candida albicans. The uptake of 3H‐uridine and 3H‐leucine as well as morphological changes in macrophages and fungus cells were studied by autoradiography. Phagocytized fungus cells remained in the yeast form and showed no activity toward incorporating 3H‐uridine and 3H‐leucine for 2 hr after infection. Cells not phagocytized began to produce germ tubes and actively incorporated both labeled precursors. Fungus cells within the macrophages produced germ tubes after 3 hr, some of which ruptured the host cells, then grew pseudohyphae and true hyphae. The uptake of both precursors was vigorous in candidal elements, but decreased in macrophages. Networks of long candidal filaments appeared 4 hr after infection and macrophages seemed to be degenerating with severely damaged synthesizing activities. When growth of C. albicans was compared in four different media, one with macrophages and the other three without macrophages, much more mycelial growth was obtained in the medium in which macrophages were present than in media without macrophages. From these results the role of macrophages in Candida infection was discussed.
Studies on the Growth Conditions of Campylobacter fetus
Authors: Miyamae Takeo
Published: November 2013

The nutritional requirement of Campylobacter fetus was investigated by static cultivation at pH 6.80 in the presence of 0.05% thioglycollate, which was added aseptically to avoid reduction of the growth‐promoting effect due to heating. Remarkable multiplication was recognized by the coexistence of l‐cystine and thioglycollate in a vitamin‐free amino acid semisolid medium. l‐Cystine was very important for growth, and the composition of a minimum number of amino acids was also required for growth, such as l‐aspartic acid, l‐glutamic acid, l‐cystine and dl‐methionine. Guanine among the nucleic acid bases brought about a distinctively heavy growth. Ribonucleic acid, ribose, glutathione, pimelic acid and a certain commercial brand of glycogen clearly showed growth‐promoting effects as demonstrated by the increase in additional amounts in a conditional amino acid medium. Similarly, taurocholate acted as an effective growth promoter out of four bile salts tested. Guanine and ribonucleic acid were detected by paper chromatography from the ingredients of media in common use for cultivation of C. fetus. A homemade yeast extract medium retained the typical cell form in almost all of the cultures during long term preservation at room temperature, in contrast to the production of various involution forms in commercial media and a meat extract medium. In conclusion, supplies of both thiol and disulfide groups fulfilled an important role by bringing into action some additives as nutritive substances to be utilized by C. fetus.
Studies on the Growth Conditions of Campylobacter fetus
Authors: Miyamae Takeo
Published: November 2013

The collaborative function of some surfactants for selective cultivation of Campylobacter fetus was examined using a plain homemade yeast extract agar medium supplemented with thioglycollate as a simple basal medium. Out of the combinations of various growth‐inhibiting substances with colistin used as the main selective growth‐inhibiting surfactant, the combined use of colistin (250 u/ml) and either desoxycholate (0.01%) or taurocholate (0.02%) in addition to brilliant green (1: 400 000–500 000) brought about a satisfactory result. There was no serious disorder of each selective inhibitory function for the growth of C. fetus and suppression to other bacteria, particularly to both Pseudomonas aeruginosa and Proteus vulgaris. It was presumed that this selectivity depended upon both the well‐balanced combination in the quantities of the surfactants and the use of a plain basal medium such as the homemade yeast extract agar.
Growth Characteristics of Bordetella pertussis in the Chick Tracheal Organ Culture
Authors: Iida Tsuyoshi , Ajiki Yoko
Published: November 2013

In the chick tracheal organ culture of Bordetella pertussis phase I organisms, marked bacterial growths were observed in the culture medium and on the tracheal fragments. However, serum‐free fresh or conditioned medium alone as well as an emulsion of tracheal tissues prepared in the conditioned medium supported little growth of the bacteria. The need for the existence of the organ fragment was inevitable. Fluorescence microscopy of the infected tracheal fragment and a time course study of bacterial growth in the medium with the organ culture suggested that the principal site of bacterial growth appeared to be on the surface of the tracheal fragments and the increase in the culture medium might be brought about mainly by the release of bacteria therefrom.
Mapping of Antibiotic Resistance Markers in Mycobacterium smegmatis
Authors: Suga Kiyoko , Mizuguchi Yasuo
Published: November 2013

Linkage relationships among loci conferring resistances to neomycin–kanamycin (nek), viomycin‐capreomycin (vic) and streptomycin (str), as well as auxotrophic markers, were analyzed employing the conjugation system of Mycobacterium smegmatis strains Rabinowitchi and PM5. Nek and vic loci were found to be closely linked to each other and also linked to argA‐6 and argB‐1. No linkage was observed between str and nek or vic. The locations of other loci, met‐5, his‐13 and leu‐11, also could not be determined. In certain recombinant types, nek and vic alleles formed stable heterogenotes. When nekr and neks or vier and vics alleles existed in one cell, sensitivity to those drugs was dominant over resistance. Segregants of nekr or vicr from the heterogenotes were detected at a frequency of about 10–4 to 10–5. Such a stable heterogenote was not detected in str allele.
Biochemical Studies on Germination of Bacterial Spores
Authors: Watabe Kazuhito , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

The inhibitory mechanism of D‐alanine on L‐alanine‐induced germination in spores of Bacillus thiaminolyticus was studied. D‐Alanine completely inhibited every L‐amino acid‐induced germinations and had the strongest inhibitory effect among all D‐isomers. A kinetic analysis of the inhibitory effect indicated that D‐alanine competitively inhibited the incorporation of L‐alanine during the initial stages of germination.
Serologic Relationship of Coxsackie A16 Viruses from the Epidemic of Hand‐Foot‐and‐Mouth Disease in Japan, 1970, to the Prototype Strain
Authors: Fujimiya Yoshiaki , Oyama Shinobu , Numazaki Yoshio , Ishida Nakao
Published: November 2013

In 1970, a great outbreak of hand‐foot‐and‐mouth disease (HFMD) due to Coxsackie A16 virus (Cox A16) occurred throughout Japan. When serologic relationship between the viruses isolated during the epidemic and the prototype strain of the serotype was examined by the tube neutralization test, the crude new isolates were found to be poorly neutralized by both an antiserum against the prototype strain and those against the isolates, although they were neutralized significantly in the plaque reduction test. However, about 2% of virus in crude suspensions of the isolates remained unneutralized even in the plaque reduction test and this fraction could be eliminated by filtering the virus materials through a 100 mμ Millipore filter. Therefore, the difficulty in neutralization of the isolates by the tube method could be accounted for by the presence of aggregated viruses. Even when filtered viruses were used, the reciprocal neutralization kinetic studies revealed a significant serological difference between the isolates and the prototype strain. Such serological properties of the isolates were not influenced by the cell types used for virus isolation or passages. All the results suggest that the Cox A16 isolates from the epidemic of HFMD in Japan, 1970, are serologically different from the prototype strain.
Genetic and Biochemical Studies on Drug‐Resistant Mutants in Mycobacterium smegmatis
Authors: Mizuguchi Yasuo , Suga Kiyoko , Masuda Kunitsugu , Yamada Takeshi
Published: November 2013

Genetic analyses of the loci conferring resistance to antibiotics known to affect protein synthesis were made employing the conjugation system of Mycobacterium smegmatis. By the complementation tests, vic (viomycin–capreomycin‐resistant) mutants were classified into two different groups; cistrons A and B. Neomycin–kanamycin‐resistant (nek) mutants, on the contrary, fall into a single cistron. Erythromycin‐resistance (ery) locus was linked to vic–nek region. The map order deduced was argB, argA, (vicA, vicB), nek, ery. Biochemical studies showed that strains resistant to streptomycin, neomycin–kanamycin or viomycin–capreomycin had altered ribosomes.
Experimental Salmonellosis
Authors: Osawa Nobutaka , Kurashige Satonori , Saito Kazuko , Mitsuhashi Susumu
Published: November 2013

An immune ribonucleic acid (RNA) preparation was obtained from the spleens of mice immunized with a live vaccine of Salmonella enteritidis. When peritoneal macrophages were infected with S. enteritidis 116–54 which had been treated by mixed cultivation with the peritoneal exudate cells of mice previously treated with an immune RNA preparation, they showed cellular resistance against the infecting bacteria. According to the results described previously and those described in this article, it can be concluded that the cellular resistance against an infection with S. enteritidis is traceable to a cellular antibody (or antibodies) detected in macrophages of mice immunized with a live vaccine of the same organism or of mice treated in vivo (or in vitro) with an immune RNA preparation.
Effect of a Lactobacillus Product Administration on the Anaerobic Intestinal Flora of Aged Adults
Authors: Yamagishi Takayoshi , Serikawa Toshihiko , Morita Ryoko , Takahashi Kentaro , Nishida Shoki
Published: November 2013

Estimations were performed on the numbers of Clostridium perfringens in 1‐ml samples of fecal specimens collected from 30 healthy aged adults. These measurements were repeated 15 times for each individual during a 16‐week period. Most of the fecal samples proved to consistently contain more than 105 C. perfringens cells and the majority of aged adults were observed to carry over 106 cells of this organism on more than eight testing occasions. Few adults were revealed to carry over 107 cells with fairly strong α‐toxigenicities without any clinical symptoms. An attempt to decrease the numbers of organisms present by continued drinking of a commercial product containing live lactobacilli resulted in failure, whilst a gradual increase in the numbers of lactobacilli and a decrease in the numbers of Bacteroides were observed. Most infants when examined for the numbers of C. perfringens in a 1‐ml portion of their fecal specimens, showed less than 102 cells of this organism and when given the Lactobacillus preparation per os exhibited a rapid increase in the numbers of lactobacilli in their intestinal contents.
Immunochemical Characterization of Cell Wall Protein Antigen Purified from the Cell Wall Autolysate of Clostridium botulinum Type A
Authors: Takumi Kenji , Kawata Tomio
Published: November 2013

The cell wall protein antigen was solubilized from the isolated cell walls of Clostridium botulinum type A by autolysis and purified by diethylaminoethyl‐cellulose column chromatography followed by gel filtration on Sephadex G‐150. The two fractions showed a high degree of the serological activity and produced a main fused precipitin line in immunodiffusion tests against the homologous antiserum. The fact that antigenic fractions contained various kinds of amino acids but no detectable amounts of amino sugars or carbohydrates suggests that the antigens were principally composed of proteins. The protein antigen possessed multiple antigenic components on immunoelectrophoresis. As serological activity, the antigen was heat‐stable and resistant to tryptic digestion but sensitive to the actions of pronase, nagarse or pepsin. The protein antigen appeared to be responsible for the common antigenicity among the proteolytic strains of C. botulinum.
An Electron Microscopic Study of Antagonism between Cephalexin and Erythromycin in Staphylococcus aureus
Authors: Nishino Takeshi
Published: November 2013

This study concerns investigations at the cellular level of antagonism between cephalexin (CEX) and erythromycin (EM) with the aid of electron microscopes and a liquid scintillation counter. Exposure of Staphylococcus aureus 209‐P to CEX and EM in combination was found to result in a marked antagonism between the two antibiotics in their effects on the growth of the organism. Observations under a scanning electron microscope revealed lysed cells in the presence of CEX alone but almost no lysis in the presence of a combination of CEX and EM. Observations under a transmission electron microscope, on the other hand, disclosed that nearly all of the cells exposed to 20 μg/ml of CEX were transformed into protoplasts with their morphological changes being most marked after 4 hr of exposure. When 1 μg/ml of EM was allowed to act alone, this exposure resulted in thickening of the cell walls. The combined use of CEX and EM, however, resulted in neither thickening of the cell walls as in the presence of EM alone nor in the formation of protoplasts as in the presence of CEX alone but merely produced the swelling of separating walls. Cellular uptake of 14C‐L‐lysine and N‐acetylglucosamine‐1‐14C into the cell wall fraction and the protein fraction was affected by CEX and EM, respectively, when used alone or in combination.
Intraleukocytic Bactericidal Activity in Patients Receiving Corticosteroid and Radiation Therapy, and in Patients with Diabetes Mellitus
Authors: Okuda Kenji , Tadokoro Ichiro , Noguchi Yoshikuni
Published: November 2013

Bactericidal activities of peripheral white blood cells obtained from patients and from healthy persons were examined in vitro. The results obtained are summarized as follows. 1. Peripheral white blood cells from patients receiving corticosteroid and radiation therapy showed decreased levels of intracellular bactericidal activities against Staphylococcus aureus. The leukocytes from almost all patients examined displayed intense activities of intracellular bacterial killing against Streptococcus pyogenes. 2. Only polymorphonuclear leukocytes (PMNs) and macrophages obtained from patients in severe stages of diabetes mellitus exhibited decreased levels of intracellular bactericidal activities against S. aureus. 3. The leukocytes from all patients examined exhibited the same levels of intracellular bactericidal effects against S. pyogenes as leukocytes from healthy persons. 4. Pseudomonas aeruginosa, which was phagocytized by PMNs obtained from healthy persons, demonstrated a remarkable degree of resistance to any intracellular bactericidal effect.
Antigenic Analyses of Lactobacillus fermenti
Authors: Shimohashi Hirotaka
Published: November 2013

Antigenic analyses of Lactobacillus fermenti were carried out by double immunodiffusion in agar using extracts prepared with cold trichloroacetic acid (TCA) or hot dilute hydrochloric acid (HCl). A common antigen of L. fermenti, designated as antigen f by the author, was extracted from whole cells with dilute HCl, but not with TCA. The antigen f was also observed in Lactobacillus casei. In addition, all strains isolated from human saliva contained antigen 6 in their cell walls, while the antigen was not observed in most of the isolates from human feces. Therefore, L. fermenti could be divided into two subgroups based upon the existence of antigen 6. Antigen 7 which was demonstrated in some strains of L. fermenti was shared by other species of lactobacilli belonging to the serological groups D and E. The common antigen 3 found in lactobacilli was extracted from all strains of L. fermenti. Sugar components of cell walls were mainly galactose, glucose and glucosamine (including N‐acetylglueosamine), but a small amount of rhamnose was present in the cell wall of only one strain. Inhibition tests with various sugars showed that the serologically active sugars were galactose for antigen f and glucose for antigen 6.
Nonspecific Elicitation of Antibody‐Forming Cells in the Mouse Spleen by Bacterial Lipopolysaccharide
Authors: Nakano Masayasu , Uchiyama Takehiko , Tanabe Masao J. , Saito Kazuhisa
Published: November 2013

Mechanisms of nonspecific elicitation of anti‐sheep erythrocyte (SRBC) hemolytic antibody plaque‐forming cells (PFC) in mouse spleens with an injection of bacterial endotoxin (lipopolysaccharide (LPS)) were studied in comparison with the genesis of naturally occurring ‘background’ PFC in normal mouse spleens and of rapidly arising PFC in mouse spleens after immunization with SRBC. The cytokinetic pattern of anti‐SRBC PFC response after an injection of LPS was quite different from that of the response elicited after immunization with SRBC. In addition, even though LPS nonspecifically elicited anti‐SRBC PFC response in mice, LPS could not confer any immunological memory on mouse immunocytes for a ‘secondary‐type’ anti‐SRBC PFC response to restimulation with LPS or SRBC. The administration of rabbit anti‐mouse thymocyte immunoglobulin or anti‐SRBC antiserum in mice markedly suppressed the PFC response after immunization with SRBC, but did not do so after stimulation with LPS. Neonatally thymectomized mice could still respond to stimulation with LPS, producing anti‐SRBC PFC in their spleens. Injections of actinomycin D or cyclophosphamide into mice resulted in obvious reductions of the PFC responses elicited by either LPS or SRBC. However, injections of these immunosuppressive antisera or drugs did not affect the number of anti‐SRBC PFC in normal mouse spleens. These results suggest that the geneses of anti‐SRBC PFC developed under different conditions, i.e., background PFC, LPS‐stimulated PFC, and antigen‐stimulated PFC, are quite different from each other, and that the nonspecific elicitation of anti‐SRBC PFC by LPS does not require the helper function of T lymphocytes. No obvious difference, however, was observed in the time of ontogenic maturation among these three different anti‐SRBC PFC in the mouse spleens judging from when they were first manifested after birth.
The Skin‐Reactive Antigens of Mycoplasma pneumoniae
Authors: Mizutani Hiroko , Mizutani Hiromichi
Published: November 2013

Guinea pigs experimentally infected with Mycoplasma pneumoniae or immunized with the organism in combination with Freund's complete adjuvant developed a delayed hypersensitive skin reaction following on intradermal injection of the M. pneumoniae antigen. The amount of protein necessary to produce the delayed skin reaction was as low as 0.01 μg. When the sonicated whole cells were extracted with aqueous acetone, the delayed skin reactivity was found mostly in the acetone insoluble (lipid‐depleted) fraction. On the other hand, the lipid fraction which was isolated by a chloroform–methanol extraction of the acetone‐soluble fraction and had a high titer of complement‐fixing activity, exhibited little delayed skin reactivity. The lipid‐depleted antigens as the whole cell antigens produced delayed skin reactivities in human patients.
Changes of Ultrastructure in Spore Coat of Bacillus thiaminolyticus during Germination and Outgrowth
Authors: Watabe Kazuhito , Kondo Masaomi
Published: November 2013

Electron microscopic observation showed that the spore coat of Bacillus thiaminolyticus consisted of at least four layers; a high electron dense outer spore coat layer with five prominent ridges, a middle spore coat layer including two layers of a high and a low electron density, and an inner spore coat layer composing six to seven laminated layers. Rapid breakdown of the cortex and swelling of the core occurred in spores which were allowed to germinate by L‐alanine for 45 min, whereas no change of surface feature was observed by scanning electron microscopy. Germination and outgrowth of spores in nutrient broth proceeded, being accompanied by morphological changes, in three steps; the first is a rapid breakdown of the cortex and swelling of the core, the second degradation of the inner layer at a prominent region of the spore coat, and the last rupture of the spore coat and emergence of a young vegetative cell.
Neutralizing Activities of Early and Late IgG Fragments from Rabbits Immunized with Herpes Simplex Virus
Authors: Shinkai Kenkichi , Yoshino Kamcsaburo
Published: November 2013

Rabbits immunized with herpes virus were bled periodically and bivalent and univalent fragments of IgG from each serum sample were prepared by enzymatic digestion. The 2‐week F(ab′)2 showed a low neutralizing activity only after addition of anti‐IgG. F(ab′)2 of the 4‐week serum retained almost all of the neutralizing activity of IgG, while its univalent fragments demonstrated none even when tested with anti‐IgG. In contrast to these early IgG fragments, univalent fragments of the 9‐week and 20‐week IgG neutralized the virus to considerable extents in the absence of anti‐IgG; after addition of anti‐IgG the activity equaled that of intact IgG in the cases of Fab′ and Fab‐II, though the activity of Fab‐I was relatively low. The three univalent fragments were all sensitive to heating at 70 C and to ultraviolet irradiation, whereas intact IgG resisted these treatments. F(ab′)2 was resistant to the heating and less sensitive to ultraviolet irradiation than univalent fragments. Neutralization kinetic curve experiments to test blocking effects of IgG fragments against the neutralization by intact IgG suggested that the early Fab′ did combine with the virus and that the late Fab′ exerted a higher blocking effect than the early Fab′.
Characterization of “Pasteurella” piscicida Isolated from White Perch and Cultivated Yellowtail
Authors: Koike Yasuo , Kuwahara Akira , Fujiwara Hiroshi
Published: November 2013

The morphological, physiological, and biochemical characteristics of twelve “Pasteurella” piscicida strains isolated from white perch and yellowtail are described and the present uncertain taxonomic status of the organisms is discussed. The organisms isolated were gram‐negative rods showing bipolar staining and pleomorphism. No spores or flagella were observed. They were non motile and viscid colonies were formed. Growth was observed in a temperature range of 20 to 30 C and the salinity range of growth was between 0.5% and 4.0%. They were aerobic and facultative anaerobic. Oxidase and catalase were produced. Nitrates were not reduced to nitrites. Lysine and ornithine decarboxylases were not produced but arginine dihydrolase was produced. The egg yolk reaction was positive. Tween 80 and tributyrin were decomposed. Phosphatase was produced. Beta hemolysis was revealed on a medium containing defibrinated blood from chickens or carp but not from mammals. Methyl red and Voges–Proskauer tests were positive, and acetoin was produced from 2, 3‐butanediol. Glucose was fermented without gas production. Acid was produced from fructose, mannose, galactose, sucrose, maltose, dextrin and glycerol. These organisms differed from all other members of the genus Pasteurella with respect to nitrate reduction, arginine dihydrolase, Voges–Proskauer and methyl red tests. The formation of viscid colonies and inability to grow in a medium without sodium chloride or at 37 C were additional characteristics of these organisms.
Chemical and Immunological Studies on the Cell Walls of Propionibacterium acnes Strain C7 and Corynebacterium parvum ATCC 11829
Authors: Azuma Ichiro , Sugimura Kazuhisa , Taniyama Tadayoshi , Aladin Aminkham A. , Yamamura Yuichi
Published: November 2013

The chemical and immunological properties of the cell walls prepared from the cells of anaerobic coryneforms, Propionibacterium acnes C7 and Corynebacterium parvum ATCC 11829, were partially investigated. The cell walls prepared from P. acnes C7 and C. parvum ATCC 11829 were composed of fatty acids, polysaccharides consisting glucose, galactose and mannose and mucopeptides consisting mainly of alanine, glutamic acid, a, ε‐diaminopimelic acid, glycine, muramic acid and glucosamine. As the fatty acid constituents of the cell wall of P. acnes C7, iso‐pentadecanoic acid and iso‐heptadecanoic acid were detected as major components. Both cell walls prepared from P. acnes C7 and C. parvum ATCC 11829 showed potent adjuvant activity on the formation of circulating antibody and development of delayed type hypersensitivity in vivo and on the primary immune response to sheep erythrocytes in vitro, however, could not augment helper function of carrier‐primed T cells and on the development of cell‐mediated cytotoxicity to mastocytoma P815‐X2 cells in C57BL/6J mice. It is also shown that the cell walls of P. acnes C7 and C. parvum ATCC 11829 act on mouse spleen cells as mitogen.
Effect of Light on Nonphotosynthetic Microorganisms
Authors: Koyama Yasumasa , Kato Fumio , Oshibi Shigenobu , Takamatsu Takeshi , Yamagishi Saburo
Published: November 2013

The effect of light on the pigmentation of various strains belonging to the genus Streptomyces was investigated. It was revealed that six species of streptomycetes, S. massasporeus, S. phaeopurpureus, S. chibaensis, S. salmonicida, S. fluvissimus and S. longispororuber, were photochromogenic, i.e., these strains can be photoinduced to synthesize pigments. On the basis of these results, pigmentation in Streptomyces is discussed.
Synchronous Cell Differentiation in Caulobacter crescentus
Authors: Iba Hideo , Fukuda Akio , Okada Yoshimi
Published: November 2013

The growth of a stalked bacterium, Caulobacter crescentus, has been synchronized easily and reproducibly by a new method. When this bacterium is grown to a late log phase in nutrient broth at 30 C with aeration, swarmer cells are accumulated in the culture to 80% of the whole cell population. When this culture is inoculated into fresh pre‐warmed broth at twentyfold dilution, it immediately initiates synchronous cell growth. Simultaneously, synchronous cell differentiation is monitored by the susceptibility of the cells to RNA phage infection. The swarmer cells accumulated in the late log phase of growth possess nearly the same susceptibility to RNA phage infection as those in the early log phase of growth while RNA phage‐adsorbing capacity is lower in such swarmer cells. It is suggested that the swarmer cells accumulated in the late log phase of growth have lost some pili.
Further Studies of Specificity of Antibodies Contained in Antiserum against Poliovirus
Authors: Urasawa Shozo , Urasawa Tomoko , Kanamitsu Masatsugu
Published: November 2013

Antigenic components of Mahoney strain (poliovirus type 1) involved in virus neutralization reaction were analyzed with mutant Mahoney strains resistant to inhibitors in equine serum (inhibitor‐resistant mutants) by means of the kinetic neutralization test. It was shown that absorption of anti‐Mahoney serum with five inhibitor‐resistant mutants yielded sera with different antibodies, of which three had distinct specificities and two specificities possibly partly related to one of those three sera. Further, it was found that stepwise selection of Mahoney variants resistant to one, two, three and four different inhibitors resulted in gradual deviation of its antigenic composition from that of the original strain. From these results, the possible presence of three or more distinct antigenic determinant sites on the surface of Mahoney strain was indicated.