bluuebunny/crossref_metadata_2025 huggingface dataset:
Initiation of the Primary Immune Response to Sheep Red Blood Cells in the Dissociated Mouse Spleen Cell Culture
Authors: Tadakuma Takushi , Watanabe Keiichi , Mitsuma Takashi , Saito Kazuhisa
Published: November 2013

From a suspension of mouse spleen cells were separated two functionally different cell types on the basis of their ability or inability to adhere to plastic dishes during a short period of incubation. Morphological observations of cells of these two fractions were made with the aid of histochemical methods. The majority of cells in the adherent fraction possessed β‐glucuronidase, which is one of the lysosomal enzymes rich in macrophages or phagocytic cells. In contrast, almost all cells in the non‐adherent fraction were devoid of this enzyme activity and identified morphologically as small lymphocytes. The adherent and non‐adherent cells were found to associate in cell clusters during the cultivation. In the cultures stimulated with sheep red blood cells, some of the non‐adherent cells which were located peripherically in the cell clusters began to show alkaline phosphatase activity in their cytoplasm. This may perhaps indicate that antibody synthesis is going on in these alkaline phosphatase‐positive cells, since in an in vivo study such cells were found to arise in the lymph nodes of immunized animals concomitantly with the appearance of specific serum antibody.
Establishment of Microorganisms Isolated from Chickens in the Digestive Tract of Germ‐Free Chickens
Authors: Morishita Yoshiyuki , Mitsuoka Tomotari , Kaneuchi Choji , Yamamoto Tetsuzo , Yamamoto Shutaro , Ogata Manabu
Published: November 2013

Germ‐free chickens were monocontaminated or dicontaminated with microorganisms isolated from the cecal contents and feces of chickens. In the case of monocontamination, the viable number of organisms in the alimentary tract considerably varied depending on the species or genus. Staphylococcus epidennidis. Streptococcus faecalis var. liquefaciens and Escherichia coli became established in much larger numbers in germ‐free chickens than in ordinary birds. While Bifidobacterium thermophilum and Catenabacterium sp. were unable to become established, other anaerobic organisms and a strain of yeast became established almost at the same level as in the ordinary microflora. Anaerobic non‐sporeforming gram‐positive rods (AR), which were occasionally found in the ordinary chickens, were always well established throughout the gut of monocontaminated birds. In the case of dicontamination, the numbers of S. epidermidis, Catenabacterium sp. and B. thermophilum were comparable to those in the ordinary microflora. Growth of S. epidermidis was suppressed by E. coli. Catenabacterium sp. and B. thermophilum became well established when given in combination with other bacteria, except that the establishment of B. thermophilum in combination with Catenabacterium sp. was irregular. Growth of AR was prevented when administered together with E. coli. On the other hand, the numbers of E. coli and S. faecalis var. liquefaciens were continuously large even in the presence of any other bacterium.
Superinfection of HVJ Carrier HeLa Cells with Ultraviolet‐Irradiated Newcastle Disease Virus
Authors: Iinuma Masao
Published: November 2013

The mechanism of enhancement of growth of partially ultraviolet (UV)‐inactivated Newcastle disease virus (NDV) in HeLa cells persistently infected with hemagglutinating virus of Japan (HVJ) (HeLaHvJ) was investigated. HeLaHVJ cells and normal HeLa cells were inoculated with UV‐irracliated NDV under similar experimental conditions, and numbers of both infective centers and NDV‐antigen producing cells in each culture were counted. It was found that the percentage of HeLaHVJ cells which produced infectious NDV or NDV‐antigen during the first cycle of infection was approximately 1.5 times higher than that of normal HeLa cells. Survival curves of UV‐irradiated NDV showed that NDV appeared more resistant to UV when titrated in HeLaHVJ cells than in assay in normal HeLa cells. Possible explanations for this phenomenon were discussed
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Yamada Hiroyasu , Takeya Kenji
Published: November 2013

Golden hamsters were used as hosts in this work, and mice of various strains as donors of antigens. 1) There were no strain differences in immunogenicity of erythrocytes from C57BL/6, AKR, SL and CF1 mice. 2) Primary intravenous immunization with mouse erythrocytes (MRBC) induced the production of hemolysin plaque‐forming cells (PFC) in a large number, but elicited only in a negligible titer production of 2‐mercaptoethanol‐resistant antibody. 3) 2‐Mercaptoethanol‐resistant antibody was produced more efficiently in hamsters pre‐sensitized with mouse lymph node (MLN) cells rather than those pre‐immunized with MRBC after a booster with MRBC. 4) Numbers of PFC in pre‐sensitized hamsters were three‐times that of the non‐sensitized hamsters after a booster with MRBC, when pre‐sensitization was performed intradermally with a small number of MLN cells. 5) Average diameter of the hemolysin plaques in pre‐sensitized hamsters was one and a half times larger than that in non‐sensitized hamsters. Conclusions agree well with the results in our previous papers that the reversed combination of hosts and antigen donors employed support the concept that certain processes required for delayed hypersensitivity contributed to antibody production under a condition suitable for antibody response.
Enhanced Synthesis of Phosphatidylethanolamine in Chick Embryo Cells Infected with Sendai Virus of Parainfluenza Type 1
Authors: Shibuta Hiroshi , Matumoto Minoru
Published: November 2013

3H‐Serine principally labeled phosphatidylserine (PS) and phosphatidylethanolamine (PE) of lipids of chick embryo cells. 3H‐Ethanolamine labeled almost entirely PE of lipids of the cells. In turnover experiment with 3H‐serine incorporated into cells before infection, the pathway from PS to PE was shown to become more active by Sendai virus infection. Similar turnover experiment using 3H‐ethanolamine revealed that PE synthesis directly utilizing ethanolamine was also slightly enhanced by the infection. The most marked difference of infected cells from control cells was the enhanced rate of 3H‐serine incorporation into lipids. These results indicated that the Sendai virus infection enhanced the metabolic pathway serine–PS–PE.
Effect of Trypsin on the Infectivity of Sendai Virus Grown in Several Host Cells
Authors: Shibuta Hiroshi
Published: November 2013

As described in a previous study, Sendai virus formed plaques in chick embryo (CE) cell monolayers with the aid of trypsin incorporated into the agar overlay medium. The virus also formed plaqůes in monkey kidney (MR) cell monolayers without the aid of trypsin. Sendai virus grown in CE cells, as well as that grown in either of other stable cell lines tested, showed much lower plaque‐forming unit (PFU) titers in MK monolayers than in CE monolayers. However, a mild trypsin treatment of the virus grown in these cells enhanced its PFU titers in MK monolayers to the level obtained in CE monolayers. In spite of this marked infectivity enhancement, the enzyme treatment little affected the hemagglutinin titers of the virus. On the other hand, Sendai virus grown in embryonated eggs or MK cells showed similar PFU titers in both CE and MK monolayers and also showed no evidence of infectivity enhancement due to the enzyme treatment.
Studies on γM and γG Antibody Response of Mice to Bovine Serum Albumin
Authors: Katsura Yoshimoto
Published: November 2013

Response of CBA mice with γM and γG antibodies to bovine serum albumin (BSA) was studied in relation to a variety of conditions of antigen administration. The variables in the conditions were doses and physical forms of antigen, and injection routes. It was realized that γG antibody response to soluble BSA and both γM and γG antibody responses to particulate forms of BSA were augmented as the dose was increased. The γM response to soluble BSA was not elevated by an increase in the amount of antigen up to 1 mg. The soluble form was not so immunogenic as the particulate forms, in which alum‐precipitated BSA was capable of inducing both γM and γG antibodies to high titers, and heat‐denatured BSA elicited preferably antibody. Alum‐precipitated BSA and the emulsified BSA were strong inducers for γG antibody response when injected subcutaneously. In any antigen form, γM response was markedly influenced by changing the injection route, the order of decreasing efficiency for antibody being intravenous, intraperitoneal and subcutaneous. The γG antibody response was hardly affected by the injection route. The effect of a single intravenous injection of 0.01 mg of endotoxin, given 1 to 2 hr after antigen injection, on γM and γG antibody production differed according to the antigen administration procedures. Generally speaking, this agent had an enhancing effect when the antigen was given in the particulate forms, and it depressed the response when the antigen was given in the soluble form.
Behavior of Lambda Bacteriophage in Nicotinamide Adenine Dinucleotide Deficient Escherichia coli Cells
Authors: Nozawa Ryushi , Mizuno Den'ichi
Published: November 2013

When niacin requiring Escherichia coli (λ) was superinfected with λ phage during development of niacin deficiency, conversion of the injected molecule to covalently closed or nicked circles decreased in accordance with the reduction of cellular NAD (nicotinamide adenine dinucleotide) content. This finding suggested that host DNA ligase, which required NAD and induced the conversion, was not functional. In niacin deficient E. coli, vegetative growth of λ occurred, indicating that there were enough materials within the deficient cells to support multiplication of the phage. However, neither induction of prophage λ nor growth of superinfecting λCI after treatment with mitomycin C was observed in E. coli (λ). This indicated that the immunity substance was active in niacin‐deficient E. coli (λ) even after treatment with mitomycin C.
Polycistronic Translation of Plant Viral Ribonucleic Acid
Authors: Kiho Yukio
Published: November 2013

Polysomes specific for tobacco mosaic virus (TMV) infection were analyzed. Two to three days after inoculation of tobacco leaves with TMV, polysomes of the large size, synthesizing TMV‐antigenic protein were observed. Molecular weight of TMV‐antigenic protein found under these conditions ranged from 2×104 to 6–8×104 daltons which might represent coat protein subunit and its trimer. Studies on the structure of polysomes bearing infecting RNA as messenger suggested that cistrons near the 3′‐end of TMV‐RNA was translated at first immediately after infection. These results favored the concept of a polycistronic translation of plant viral RNA, in contrast to a monocistronic translation observed in the mammalian system.
Studies on the Adjuvant Action of Mineral Oil and Bacterial Endotoxin
Authors: Sohmura Yasunobu
Published: November 2013

Water‐in‐oil emulsion (WOE) of the Freund's type and a bacterial endotoxin (ET) enhanced the antibody response of mice to bovine γ‐globulin (BGG) in a different manner. WOE even without the antigen revealed an adjuvant action when given prior to or simultaneously with the antigen, while ET was effectual when given simultaneously with or after the antigen. Thus, the concurrent administration of these two adjuvants either before or after the antigen secured enhancement. It was shown that ET facilitated IgM antibody formation. WOE including antigen (BGG‐WOE) was found to form an ‘antigen‐depot’ at the injected site. Antigen released bit by bit from this depot thus might supply a continuous stimulus for the antibody response. This was mimicked by a divided daily injection of a small amount of antigen without adjuvants. Surgical removal of the hind foot containing the depot resulted in reduction of the circulating antibody. The popliteal lymph node cells from mice given BGG‐WOE via hind foot pads could adoptively immunize X‐irradiated recipients without the additional administration of antigen, axillary lymph node cells and spleen cells being unable to do so. ET was inadequate for this purpose. The morphological changes of the nodes seemed compatible with these results.
Magnesium‐Stimulated Resynthesis of Osmoresistant Layer by Candida albicans Spheroplasts
Authors: Ota Fusao
Published: November 2013

Spheroplasts of Candida albicans could be obtained by treating cells with a snail enzyme in the presence of 0.75 m MgSO4 as a stabilizer and cells could subsequently be regenerated by the thin‐layer‐agar plating method. The initial regeneration process was affected by stabilizers such as MgSO4, MgCl2, MnSO4, CaCl2, KCl, NaCl and sucrose. It was found that 0.68 m MgSO4 was the best stabilizer. Good stabilization was also achieved in any medium with 0.5–0.75 M sucrose, although stabilizing agents were in general most effective at concentrations giving about 22 atmospheres of osmolality. This osmolarity was lower than that used to prepare spheroplasts. Using this thin‐layer plating method at least three factors were essential for resynthesis of a wall layer: (1) a carbon source, (2) a stabilizer and (3) several minerals. Nitrogen in the form of an amino acid stimulated formation of the wall layer. Relationship between these factors and osmolarity of the medium during regeneration of the wall layer is discussed.
Mechanism Determining the Host Specificity of Tobacco Mosaic Virus
Authors: Kiho Yukio , Machida Haruhiko , Oshima Nobuyuki
Published: November 2013

In order to know the mechanism controling the host specificity of tobacco mosaic virus (TMV), three species of plants showing various degrees of resistance to TMV or TMV‐RNA infection were selected and the fate of infecting viral genome was studied. Extract was obtained from leaves 0.5–6 hr after inoculation of 32P‐TMV or 32P‐TMV‐RNA and analyzed by sucrose density gradient centrifugation. It was found that polysomes containing infecting 32P‐RNA were formed in plants resistant to TMV to the same extent as in susceptible tobacco plants, suggesting that the host specificity of TMV is determined at a stage of viral multiplication later than the step of translation of infecting viral genome.
Extracellular Crystalline Lattice Material of Corynebacterium diphtheriae Revealed by Electron Microscopy
Authors: Kawata Tomio , Masuda Kuniyoshi
Published: November 2013

Crystalline material showing square lattice arrays attached to the cell envelope of Corynebacterium diphtheriae grown in liquid cultures was demonstrated under the electron microscope using negative staining and sectioning techniques. Two types of lattice patterns could be found in the crystal. One was a square lattice consisting of parallel lines (about 3.2 nm in diameter) with an average distance between their centers of 5.3 nm. The second type of lattice pattern was composed of thinner parallel lines (approximately 2.2 nm in diameter) with a closer periodicity being about 3.5 nm. The two types of lattice patterns appeared to intersect each other at an angle of 45 degrees. The lattice arrays were disorganized and disappeared upon treatment with proteolytic enzymes, alkali and sodium dodecyl sulfate. This crystalline material seemed to be composed of protein(s) but not to be related to diphtheria toxin.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Makidono Reiko , Takeya Kenji
Published: November 2013

It was confirmed by passive transfer experiments that the function of thymus‐derived cells specific for hamster erythrocytes (HRBC) was deficient in the low‐responder mouse strains. 1) Antibody production against HRBC was enhanced by passive transfer of thymus cells from normal SL mice (high‐responder) to normal C57BL/6 mice (low‐responder). 2) The enhancing effect of passive transfer of thymus cells from SL mice was abrogated by pre‐sensitization of the recipients (C57BL/6) with thymus cells from SL mice. 3) In C57BL/6 mice, antibody production against HRBC was enhanced by the transfer of lymph node or spleen cells from C57BL/6 mice which had been sensitized with HRBC in Freund's complete adjuvant or hamster lymphoma cells.
Isolation and Characterization of Spontaneous Mutants of Staphylococcus aureus S6 Possessing Quantitative Differences in Enterotoxin B Production
Authors: Kondo Masaomi , Katsuno Shingo
Published: November 2013

Spontaneous mutants possessing quantitative differences in enterotoxin B production were isolated from the parent strain of Staphylococcus aureus S6 by using NZ‐amine agar plate which contained anti‐enterotoxin B antiserum. A shake‐flask culture of S. aureus S6 715H, a high enterotoxin B‐producing strain, extracellularly produced 370 μg/ml of enterotoxin B after 16 hr of incubation at 37 C. A shake‐flask culture of S. aureus S6 715NH, a low enterotoxin B‐producing strain, produced 22 μg/ml of enterotoxin B under the same conditions. The difference in the ability to produce enterotoxin B among the strains resulted from differences in their activities of de novo enterotoxin B synthesis. A quantitative difference in the production of individual extracellular proteins existed in the strains. S. aureus S6 715H produced higher levels of α‐and δ‐hemolysins, alkaline phosphatase, ribonuclease and staphylokinase (fibrinolysin) than S. aureus S6 715NH. These results suggest that the production of enterotoxin B and most of the other extracellular proteins, at least the above five toxins or enzymes, may be dependent upon some coordinated control mechanisms in S. aureus S6.
Ampicillin‐Resistant R Factors Derived from Shigella Strains
Authors: Tanaka Tokumitsu , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Sixteen Shigella strains resistant to ampicillin(APC) were isolated from 3760 strains examined in 1965. Five strains among these isolates were resistant to tetracycline(TC), chloramphenicol (CM), streptomycin(SM) and sulfanilamide(SA) in addition to APC, and transferred their APC resistances by conjugation together with the aforementioned four drugs. The 5‐drug resistance was transferred to an Escherichia coli strain and the resistance was analyzed by both conjugation and transduction. Three out of the five Shigella strains transferred a single type of R(TC.CM.SM.SA.APC) factor, but the linkage relationship between the resistant markers were different from each other. The ampicillin gene, however, was always linked close to the genes governing both transfer and replication. The 5‐drug resistance was transferred to an E. coli strain from the remaining two Shigella strains. According to the genetic studies of E. coli strains which had acquired the 5‐drug resistance, it was concluded that the two parent Shigella strains carried two types of R factors in a cell, i.e., fi+R(TC.CM.SM.SA) and fi‐(SM.APC). These two R factors in a cell sometimes recombined and formed a recombinant R factor possessing the 5‐drug resistance in which the ampicillin gene was linked close to the transfer gene. The enzymological properties of the five penicillinases coded by R factors possessed by the five Shigella strains were all similar to each other and of the type 1 penicillinase, which was demonstrated from Klebsiella pneumoniae and mediated by most of the R factors.
Genetic Structure of an R Factor Conferring Ampicillin Resistance
Authors: Hashimoto Hajime , Tanaka Tokumitsu , Mitsuhashi Susumu
Published: November 2013

An R factor carrying resistant genes, ampicillin (amp), tetracycline (tet), chloramphenicol (cml), dihydrostreptomycin (sir), and sulfanilamide (sul), was analyzed by three genetic techniques: spontaneous segregation, transduction, and recombination. The R factor was stable in Shigella and Escherichia coli, but in Salmonella the (cml.str.sul) genes were spontaneously lost at a high frequency. Almost all of the transductants with Pl contained the five drug markers, but a very small number of them carried both tet and amp genes, or the (amp.cml.str.sul) genes. In conjugal transfer from a strain carrying a single type of R factor to R+ recipient, the recombination took place between the amp and tet, amp and cml, and between the cml and str genes, at a frequency of 23.5, 39.5, and 3.0%, respectively. Crossover between the amp and transfer (tra) loci was not observed. A circular genetic model is proposed in which the genes are arranged in the order –tet9–tet1‐tra.amp–cml–str–.
Cephalexin‐Induced Morphological Alterations in the Surface Structures of Staphylococcus aureus and Escherichia coli Demonstrated by Scanning Electron Microscopy
Authors: Nishino Takeshi , Nakazawa Shozo
Published: November 2013

The scanning electron microscope was used to investigate the alterations in surface morphology of Staphylococcus aureus 209P and Escherichia coli NIH induced by the action of cephalexin known to interfere with cell‐wall synthesis. Exposure to cephalexin produced a series of changes on the surface morphology in proportion to the concentrations of cephalexin added. Untreated S. aureus cells had smooth contours. Exposure to 1 μg/ml of cephalexin during the logarithmic phase of growth in S. aureus did not produce any detectable changes. Upon exposure of S. aureus to 5 μ/ml or 10 μg/ml, some cells were larger than normal and showed abnormal cell division‐like structures in part. When S. aureus was exposed to 50 μg/ml, cell division was completely inhibited, and no formation of grape‐like clusters was observed. Untreated E. coli cells appeared to have smooth and regular contours. E. coli propagated almost normally upon exposure of the organisms to 1 μg/ml of cephalexin. Filamentous structures were observed with the exposure of E. coli to 12.5 μg/ml or 25 μg/ml, but spheroplast‐like structures were not observed. Exposure to 100 μg/ml of cephalexin resulted in the formation of marked filamentous cells and spheroplast‐like structures having multiple small saccular outpouchings. Scanning electron microscope demonstrated more completely the morphological abnormalities induced by cephalexin.
Transformability of Group H Streptococcus Challis
Authors: Ito Takako , Hirano Toyo , Tomura Tsuneko , Yoshioka Morimasa
Published: November 2013

Two types of hemolysis, α and β were formed by group H streptococcus wild type strain Challis on horse blood agar, and both of the hemolytic markers could be transformed in each. α‐Hemolysis‐acquired transformants lost the competence‐provoking factor producibility (CPF–). When DNAs, obtained from strains Challis α, dihydrostreptomycin sulfate‐resistant Challis α, and Wicky, were used CPF– transformants appeared in high frequencies. DNAs from incompetent β cells grown in ET3 medium for 6 hr or in heated ET3 medium for 2 hr also transferred the CPF–. CPF producibility (CPF+) was transformed with DNA from competent Challis β, while it was not transformed to Challis β with DNA from Wicky cells in competent and incompetent phases. Among β transformants obtained there were (1) those having CPF+ and competency, (2) those having competency but CPF–, and also (3) those defective in both. Based upon these results, the transformability is discussed regarding close relation to DNA receptor site or CPF attachment and also on the probable transfer of itself by transforming DNA.
The Incidence of R Factors in Bordetella bronchiseptica Isolated from Pigs
Authors: Terakado Nobuyuki , Azechi Hayami , Ninomiya Kiyoji , Fukuyasu Tsuguaki , Shimizu Takeshi
Published: November 2013

Bordetella bronchiseptica strains isolated from the nasal cavities of young pigs in Japan from 1969 to 1972 were surveyed for drug resistance and distribution of R factors. Of 304 strains examined, 71 (23%) were resistant to either one or more of following three drugs, streptomycin (SM), sulfadimethoxine (SA), and aminobenzyl penicillin (APC). Triple (SM.SA.APC)‐resistance was most frequent among these resistant strains. Strains of double (SM. SA)‐ or single (SM)‐ and (SA)‐resistance were also isolated, but were very few in numbers. Of the 71 drug‐resistant strains, 61 (86%) were found to carry R factors which were capable of conjugal transfer. All of these R factors had the triple (SM.SA.APC)‐resistant markers and were identified as fi– (no fertility inhibition) type. The (SM.SA.APC)‐resistant strains carrying R factors had been isolated from pigs reared on various farms in different districts, and consequently the prevalence of B. bronchiseptica strains carrying R factors was considered to be relatively wide‐spread in young pigs.
Studies on the Growth Conditions of Campylobacter fetus
Authors: Miyamae Takeo
Published: November 2013

The optimal growth pH and conditions necessary to give a full manifestation of the effect of reductants upon the growth of Campylobacter fetus were investigated using static cultivation. The multiplication of cells was limited to a narrow range of added reductants. Thioglycollate supplemented without heating produced heavy growth in combinations of 0.05% at pH 6.80 or 0.1% at pH 6.60, and 0.2% at pH 7.00 or 7.20. Although the stimulating effect depended upon the time between the addition of the supplement and inoculation, two prominent peaks in the growth curve always appeared with both alkaline and acidic pH ranges of the medium. Autoclaving the medium with some SH compounds brought about this growth only at around pH 6.50. Consequently there was a uniform decrease in the initial pH. l‐Cysteine and ascorbic acid also gave similar effects. Glucose remarkably prevented the growth promotion due to SH compounds. Optimal initial reduction potential for growth in a semisolid medium was considered to be −0.05 to −0.08 volt at an initial pH of 6.60. SH compounds supplemented without heating brought about a marked longevity of the culture in comparison with a culture in an autoclaved medium of the same composition. For the growth of C. fetus in semisolid medium, it was important to avoid decreasing the growth‐promoting activity of some of the supplemented SH compounds by not heating but adding the supplement aseptically. Also a rigid regulation of the pH, the concentration of the added reductants, and the time of inoculation into the fresh medium were important.
Isolation of a Common Cell Wall Antigen from the Proteolytic Strains of Clostridium botulinum
Authors: Takumi Kenji , Kawata Tomio
Published: November 2013

A protein‐rich antigen was extracted from the cell walls of Clostridium botulinum type A strain 190L by relatively mild treatments with detergents, tryptic digestion or cell wall autolysis. The cell wall antigen was relatively labile, and destroyed by acid or formamide hydrolysis. Immunodiffusion studies with the trypsin‐treated cell wall extracts from various strains of C. botulinum indicated that the antigenic cell wall polymers among the proteolytic strains of types A, B and F but not among the nonproteolytic strains of C. botulinum shared a common antigenic specificity.
β‐Lactamase Formation and Resistance of Proteus morganii to Various Penicillins and Cephalosporins
Authors: Yaginuma Satoshi , Sawai Tetsuo , Yamagishi Saburo , Mitsuhashi Susumu
Published: November 2013

Ten strains of Proteus morganii were selected and the production of β‐lactamase was studied. The cellular level of β‐lactamase from P. morganii was regulated by the concentration of the inducer in the growth media, and the enzyme activity appeared within 10 min while its highest value was obtained 2 hr after the addition of benzylpenicillin as the inducer. The resistance level to cephaloridine was generally related to the amount of β‐lactamase activity among the ten strains of P. morganii, but no relation was found between the ampicillin susceptibilities and the amounts of β‐lactamase in their cell‐free preparations. The maximum rate of hydrolysis of cephalosporin C and several other derivatives of 7‐aminocephalosporanic acid by a crude enzyme was five times higher than that of benzylpenicillin. Semisynthetic penicillins were resistant to hydrolysis and exhibited competitive inhibition. Among the semisynthetic penicillins, dicloxacillin and carbenicillin were powerful competitive inhibitors of the β‐lactamase from P. morganii.
A Capillary Tube Method for Counting Viable Cells of Bifidobacterium bifidum Grown in a Solid Medium1
Authors: Nakamura Hiroshi , Samejima Keijiro , Tamura Zenzo
Published: November 2013

A method which permitted counting viable cells of Bifidobacterium bifidum N4 in a solid medium was developed. A piece of the solid medium (0.7 ml) was quantitatively obtained with the aid of an agar‐puncher device and was homogenized in a Potter–Elvehjem homogenizer after the addition of 9.3 ml of sterile physiological saline. A 10‐fold dilution of the homogenate was repeated several times to make a series of dilutions. An aliquot (0.2 ml) of the appropriate dilution was used for counting the viable cells using a capillary tube method. The accuracy and the reproducibility of the method were comparable with those of the conventional plate counting method. By using established procedures the behaviors of B. bifidum N4 in a solid medium were studied. Viability of the organism in a solid medium lacking an energy source (lactose) was generally correlated to the period of preculture; the longer the period of preculture, the shorter was the span of cell life.
Biochemical Studies on Germination of Bacterial Spores
Authors: Watabe Kazuhito , Ichikawa Tomio , Kondo Masaomi
Published: November 2013

The initiating mechanism in the germination of Bacillus thiaminolyticus spores was studied with 14C‐L‐alanine. A characteristic pattern of incorporation of L‐alanine into the spores was observed during the early stages of germination with two incorporation peaks, one occurred just after contact with L‐alanine (first incorporation) and the other 5 min later (second incorporation). L‐Glutamine, L‐valine, or L‐serine substituted for the incorporation of L‐alanine during the first stage of germination. Although, L‐alanine taken up during the first incorporation phase was extractable with trichloroacetic acid (TCA), that taken up during the second incorporation phase was not extractable. The distribution of radioactivity showed that incorporated L‐alanine was located in the spore coat, mainly in the paracrystal fraction. The radioactive material which remained in the germination medium or was extractable from the spore coat fraction with TCA treatment or pronase digestion was identified as alanine. Significance of incorporation of L‐alanine and its location in the spore in reference to the initiation of germination is discussed.
Model System for the Adsorption of Tobacco Mosaic Virus
Authors: Kiho Yukio
Published: November 2013

Materials which can adsorb tobacco mosaic virus (TMV) were isolated from tobacco leaves and studied for applicability as a model system for TMV adsorption. Leaves were homogenized and fractionated by sucrose density gradient centrifugation. One fraction adsorbed TMV in the presence of polyornithine. Deduced from its sensitivity to trypsin and detergent as well as from its manner of isolation, the material responsible for adsorption of TMV seemed to be cytoplasmic membrane. Membrane derived from light particulate, as well as cytoplasmic membrane, seemed to be capable of adsorbing TMV. Shorter rods obtained by sodium dodecyl sulfate or sonic treatment of TMV could adsorb to membrane as efficiently as TMV. Viral protein subunit could not adsorb whereas helical rods made of viral protein aggregates could. A two‐step nature of the adsorption of TMV was suggested: a salt‐sensitive and a subsequent salt‐resistant steps. In the first step, ionic bonding plays a main role in the combination between TMV and membrane. Adsorption of 14C‐labeled TMV was inhibited by an excess amount of non‐labeled TMV or cucumber green mottle mosaic virus but not by potato virus X or rice dwarf virus, suggesting the specific nature of adsorption. In contrast to the observed specificity on the part of virus, a membrane fraction isolated from various plants, including non‐hosts for TMV, could adsorb TMV. This may imply that adsorption and injection are not the determinant of host specificity in plant viral infection.
Effect of Calcium on the Cell Infectivity of Virulent Shigella flexneri 2a
Authors: Osada Yasuaki , Une Tsutomu , Ikeuchi Tora , Ogawa Hidemasa
Published: November 2013

Effect of calcium ions (Ca++) on the virulence of Shigella flexneri 2a was examined with reference to its infectivity to cultured cells, the guinea pig eye, and the ligated small intestine of rabbits. The organism grown in a calcium‐containing medium showed a significantly higher ability to penetrate HeLa cells than that of organism grown in a calcium‐deficient medium. This ability was constantly maintained in the presence of Ca++, while readily lost in the absence of Ca++. Similarly, its pathogenicity for the guinea pig eye and the ligated small intestine of rabbits was more or less greater in cases with Ca++ than in cases without Ca++. Ca++ seemed to play an important role in bacterial adhesion to the cell membrane, by which the cellular engulfment was induced. The presence of Ca++ in the environment of bacterial growth was emphasized on development and maintenance of virulence of S. flexneri 2a.
Drug Resistance and Distribution of R Factors among Escherichia coli Strains
Authors: Tanaka Tokumitsu , Kobayashi Akio , Ikemura Kengo , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Three hundred and thirty‐eight strains of Escherichia coli from clinical sources were examined for their drug resistances and R factors in 1971. With reference to four drugs, i.e., tetracycline(TC), chloramphenicol(CM), streptomycin(SM) and sulfanilamide(SA), the SA‐resistant strains were isolated most frequently followed by SM‐, TC‐ and CM‐resistant ones in decreasing order. Among those resistant to the aforementioned four drugs, 20.3, 19.0, 7.7 and 18.3% were single‐, double‐, triple‐ and quadruple‐resistant, respectively. The R factors were demonstrated at a frequency of 29.2% among these resistant strains. From the strains possessing quadruple‐resistance, the conjugationally transferable R factors were demonstrated most frequently (60.5%) followed by triple‐ (59.1%), double‐ (21.7%) and single‐resistant strains (4.5%). The isolation frequencies of strains resistant to kanamycin (KM) and ampicillin (APC) were 1.5 and 4.7%, respectively; being fewer in numbers compared with those resistant to the four drugs. Strains resistant to cephaloridine, nalidixic acid, Colistin, gentamicin C complex and furatridine were not isolated. Strains resistant to KM and APC were demonstrated mostly from those possessing multiple resistance with reference to the aforementioned four drugs. Among the strains carrying either APC‐ or KM‐resistance, 76.9 and 100% respectively, were found to carry R factors, indicating that most of the APC‐ and KM‐resistances were conferred by the R factors. Twenty percent of the R+ strains with reference to resistance to the four drugs carried two types of R factors in a cell, being in a hetero‐R state, and 66.7% of the strains carrying R factors with APC‐ or KM‐resistance were in a hetero‐R state.
Studies on Complement‐Fixation Reaction in Equine Infectious Anemia
Authors: Nakamura Junji , Kuroda Sadao , Ueda Susumu , Samejima Togo
Published: November 2013

Complement‐fixing (CF) and complement fixation‐inhibiting (CFI) antibodies were investigated sequentially in three horses infected with equine infectious anemia (EIA) virus. The CF activity was first demonstrated 6 or 8 days after onset of the first pyrexia. The CFI activity developed a short period later, and caused a decrease of apparent CF titer of the whole serum. However, both antibodies tended to increase with advance of the disease course in two persistently infected horses, whereas they became completely undetectable during the late‐stage in the remaining horse which showed no evidence for recurrence of pyrexia or persistence of viremia. The CF activities determined with varying dilutions of serum were distinctly different in pattern between the early‐stage serum having the CF activity alone and the late‐stage serum having both the CF and the CFI activities. The CF antibody was precipitated by 27–30% saturation with ammonium sulfate (SAS) while the CFI activity distributed in a wider range of precipitates formed by 26–32% SAS. The CFI activity was demonstrated to a higher titer when a relatively small amount of antigen was sensitized with CFI antibody prior to addition of reference CF antibody than when the three reagents were mixed at one time. The late‐stage serum with a strong CFI activity against EIA antigen had no effect on the CF reaction of other viral antigen–antibody systems such as equine influenza and equine rhinopneumonitis.
Interferon Production and Resistance to Viral Infection Induced by Poly I · Poly C in Chick Embryo Cells
Authors: Machida Haruhiko , Kuninaka Akira , Yoshino Hiroshi
Published: November 2013

Two different growth media, one based on Eagle's minimum essential medium (MEM) and the other on Earle's balanced salt solution–lactalbumin hydrolysate–yeast extract (YLE), were used for growing primary chick embryo cells (CEC), and resistance to viral infection and interferon production induced by polyinosinic‐polycytidylic acid (poly I·poly C) were compared. In CEC grown in Eagle's MEM, treatment with poly I·poly C at a concentration as low as 1.0 ng/ml was sufficient to induce a detectable resistance to infection with vesicular stomatitis virus (VSV), while more than 300‐fold concentrated poly I·poly C was required to induce a similar resistance when the cells were grown in YLE. The cells grown in YLE did not produce an appreciable amount of interferon, whereas a significantly higher level of interferon was produced by the cells grown in Eagle's MEM. A similar phenomenon was observed in the interferon production of chick embryo cells treated with ultraviolet light (UV)‐irradiated Newcastle disease virus (NDV) and in the induction of resistance to vaccinia virus in cells treated with poly I·poly C. It was found that the response of cells, bathing in one growth medium, to poly I·poly C was not affected by replacing it with the other at the same time with the addition of poly I·poly C, and that the response of CEC was strongly dependent upon the medium used for cultivation. These facts suggested that the observed difference in the response of cells to poly I·poly C was not due to a direct interaction between the inducer and medium components but to the physiological state of CEC established during their growth. Which component of YLE was responsible for such a lowered response of cells to poly I·poly C was also examined, and the marked reduction of PDD50 by the replacement of lactalbumin hydrolysate of YLE with amino acids and the increase of PDD50 by addition of lactalbumin hydrolysate to MEM suggested that lactalbumin hydrolysate might play an important role in this phenomenon.
A Rapid and Simple Method for Assaying Interferon1
Authors: Suzuki Jiro , Akaboshi Tomoko , Kobayashi Sigeyasu
Published: November 2013

A new procedure of assaying interferon (IF) has been developed. Cell suspension was dispensed into liquid scintillation counting vials together with IF sample. During an overnight incubation, the cells adhered sufficiently to the bottom of the vials and all the subsequent procedures were carried out without transfer of the cells from the vials. Vesicular stomatitis virus was inoculated and virus‐specific RNA was labeled by adding 3H‐uridine and actinomycin D to the medium. Incubation was terminated prior to completion of a single‐step growth of the virus and radioactivity of the labeled cells in each vial was determined. The reciprocal of the IF dilution which reduced the radioactivity in viral RNA by 50% was taken as the titer. The present procedure consists of simple manipulations and can be completed within 24 hr. Furthermore, it is quite reproducible and gives a titer almost identical to that obtained by the conventional plaque‐reduction dose method. The procedure can be applied to mouse L cells, rabbit RK‐13 cells and human FL cells, without modification.
Microbial Adjuvant and Autoimmunity
Authors: Nakashima Izumi , Kato Nobuo
Published: November 2013

With the use of the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) as a powerful adjuvant, high precipitin responses could be induced in mice to syngeneic eyeball extracts and thyroid gland extracts which were normally nonimmunogenic. Only very weak responses were induced to eyeball extracts by Freund's complete adjuvant. Repeated administrations of the antigens mixed with CPS‐K at time intervals of 30 days (more than twice for the eyeballs or more than three times for the thyroid glands) were required for induction of high precipitin responses. Antibody responses detectable by the immunofluorescent technique could be induced to syngeneic lymphoid tissue extracts by injecting the mixture of antigen and CPS‐K more than five times at time intervals of 30 days. These findings suggest that repeated stimulation by autoantigens together with such a strong adjuvant as CPS‐K can terminate natural tolerance against autoantigens.
Arginine Gene Cluster of Serratia marcescens
Authors: Matsumoto Hideki , Hosogaya Shigemi , Suzuki Kiyoshi , Tazaki Tadakatu
Published: November 2013

Biochemical and genetic studies on the arginine‐requiring auxotrophs derived from a Serratia marcescens strain were carried out. The arg mutants were classified into seven biochemical groups based on their growth response to five precursors of arginine biosynthesis and enzyme deficiency. Reciprocal transduction tests among those arg mutants divided them into three linkage groups, and the fine mapping in each of the groups by two‐ or three‐point crosses revealed the following arrangement of loci. (1) arg44–thy11–lys1; (2) met1–glt2–argE–(arg19–arg51)–arg120–argG–argH; (3) arg33–pyr4. Five of the seven biochemically distinct arg mutants belonged to the second linkage group, and they constituted an arg‐gene cluster. A characteristic feature of the arg‐gene cluster of S. marcescens is that it involves argG, which was previously reported only in the Proteus group of Enterobacteriaceae.
Staphylococcal Footpad Infection in Mice
Authors: Katagiri Ken , Hori Kenji , Nishino Takeshi , Fukao Takashi
Published: November 2013

Local footpad infection in mouse was investigated with 55 clinically isolated strains of Staphylococcus aureus. When 107 viable cells were inoculated into the footpad, local swelling and bacterial growth resulted after 24 hr. With a dose of 106 cells, moderate swelling was observed after a few hours but the reaction had almost disappeared after 24 hr. About 75% of the staphylococcal strains tested caused footpad edema in mice at doses of 107 cells. A statistical comparison of the virulence of the organisms on intravenous and intraperitoneal injection with that in inducing footpad swelling is also reported.
Requirement of Newly Synthesized Protein for the Priming Activity of Interferon
Authors: Ito Fumiaki , Suzuki Jiro , Kobayashi Sigeyasu
Published: November 2013

Pretreatment of mouse L cells with interferon (IF) enhanced IF production in response to polyinosinic‐polycytidylic acid (poly I·poly C). Post‐treatment of cells with IF caused no significant enhancement of IF production. The enhancing effect of IF pretreatment (priming) reached a maximum after incubation with IF (10 or 100 units/ml) for 4–6 hr at 37 C, but this effect was absent when the incubation was done at 4 C. Cells which were incubated for additional several hours at 37 C after IF pretreatment at 4 C did not develop the primed state nor the antiviral state. The presence of protein synthesis inhibitors during the IF pretreatment depressed, though not completely, the development of the primed state. The residual priming effect was lost when the cells were incubated with the inhibitors at 37 C for 2 hr before they were exposed to poly I·poly C. There was no significant difference in the binding rate of poly I·poly C to cells between IF‐treated and untreated cells. The degradation rate of cell‐bound poly I·poly C and its sensitivity to exogenous pancreatic ribonuclease in the pretreated cells were also similar to those in the untreated cells.
Inhibiting Materials for γ Phage Adsorption to the Cell Wall of Bacillus anthracis, Strain Pasteur No. 2‐H
Authors: Watanabe Takashi , Shiomi Toshiro
Published: November 2013

Cell wall preparations of Bacillus anthracis, strain Pasteur No. 2‐H, were treated with heat or with acetone and ether. Both of the treated cell walls preparations inactivated γ phage. The centrifuged supernatant of the heat‐treated cell walls was fractionated on Sephadex G‐200, and four fractions containing reducing sugars were obtained. The first fraction had the phage‐inactivating activity. On the other hand, the fourth fraction had no phage‐inactivating activity, but strongly inhibited phage adsorption to the cell walls. In the fourth fraction, glutamic acid, alanine, 2, 6‐diaminopimelic acid and glucosamine were detected by paper chromatography after acid hydrolysis. Authentic D,L‐2, 6‐diaminopimelic acid and D‐glucosamine markedly inhibited phage adsorption to the cell walls. D‐Galactosamine, D‐mannosamine and L‐lysine also showed similar activities. Results suggest the possibility that one or a combination of these substances defines the characteristics of phage adsorption to the cell walls of B. anthracis, strain Pasteur No. 2‐H.
Comparative Studies on Generalized Transducing Bacteriophages of Proteus mirabilis, φm and π1
Authors: Nakamura Motoko , Horiuchi Sankichi , Nakaya Rintaro
Published: November 2013

Comparative studies were made on the generalized transducing bacteriophages of Proteus mirabilis φm (Nakaya and Rownd), π1 (Böhme), and a clear plaque‐forming mutant φm‐c, derived from φm. Electron microscopic observations revealed that these phages were morphologically identical, indicating that they belonged to the group C of Bradley's classification, or to the type Cl of Ackermann's classification. Phages φm and π1 formed characteristic turbid plaques different from each other, and the plaques of π1 were smaller in size than those of φm. The plaques of phage φm‐c were clear and also were the largest in size among those studied. Average latent periods of φm and π1 were 70 and 60 min, respectively. Average burst size was found to be 30 and 10 plaque‐forming units per infected cell for φm and π1, respectively. It was confirmed by cross neutralization tests that φm and π1 differed serologically from each other. The host range of the two phages also differed, and phage φm was more sensitive to heat than π1. These results indicate that phages φm and π1 are different types of phages. Majority of the properties of phage φm‐c were nearly identical with those of phage π1 except that the multiplication of φm‐c was more strongly inhibited by methylene blue than that of φm and π1. Phage φm‐c is considered to be a clear mutant of φm.