bluuebunny/crossref_metadata_2025 huggingface dataset:
Bovine Adenovirus
Authors: Tanaka Yoshio , Inaba Yuji , Ito Yasuichiro , Omori Tuneyoshi , Matumoto Minoru
Published: November 2013

Two adenovirus strains were isolated in calf testicle cell cultures from blood specimens of cattle in Japan. This is the first isolation of bovine adenovirus reported in Japan. The isolates were antigenically similar to each other and distinct from the hitherto described serotypes 1, 2 and 3 of bovine adenovirus. Unfortunately, bovine adenovirus types 4 and 5 were not available for comparison, and hence, until the matter is settled, the virus will be called “Bovine adenovirus type Nagano”. Nagano virus was identified as adenovirus on the bases of the inhibitory effect of 5‐iodo‐2′‐deoxyuridine on virus replication, ether‐resistance, effect of temperature and pH on infectivity, and fine structure of the virus particle. The virus grew and formed intranuclear inclusion bodies, a characteristic of adenovirus, in bovine testicle cells but not in bovine kidney cells. The virus agglutinated rat erythrocytes very poorly, but not sheep, goat, cattle, horse, guinea pig, hamster, chicken, and mouse cells. The virus produced adenovirus group‐specific antigen in cell cultures. Sero‐negative calves were readily infected with the virus by the intravenous, subcutaneous, oral or intranasal routes of inoculation. The infected animals produced antibodies and showed a mild clinical reaction comprised of rhinorrhea, diarrhea and a degree of pyrexia; low‐titered viremia of short duration and leukopenia were also observed. A serologic survey indicated wide‐spread dissemination of the virus among Japanese cattle, but further studies are needed to determine the etiologic significance of the virus in the natural disease in cattle.
Bacteremia in Experimental Tyzzer's Disease of Mice
Authors: Takagaki Yoshio , Fujiwara Kôsaku
Published: November 2013

Bacteremia was observed during the late stage of experimental Tyzzer's disease in mice. The number of organisms in the blood in mice treated with cortisone increased markedly in the infection with highly virulent organisms, whereas bacteremia was of low incidence and less severe in infections with low virulence organisms. The number of organisms in the blood stream was shown to increase linearly during the course of fatal infection attaining a maximum level of 107 organisms per ml blood. The number of organisms in the blood was found to be closely related to the number in the liver when bacteremia was observed. In the peripheral blood, organisms were first detectable when the number of organisms in the liver gained a level of 107 per g tissue, and the subsequent increase in the number of bacteria in the blood was approximately 3 times more rapid than in the liver. The organisms in the blood were comparable to those in the liver morphologically as well as in pathogenicity. Histopathological examination frequently revealed liberation of organisms from liver cells into sinusoids. There was no evidence of significant multiplication of the organisms in organs other than the liver.
Subgroups in Group III of RNA Phages
Authors: Miyake Tadashi , Shiba Tadayoshi , Watanabe Itaru
Published: November 2013

In our previous studies, RNA phage strains were separated into 3 major groups on the basis of filtration efficiency through Millipore filters. In the present study, the strains of group III were shown to be further divided into 3 subgroups: (a) Qβ, NH, SG; (b) VK, SO; (c) ST. This subgrouping was found to be compatible with the serological grouping and pronase sensitivity with the exception of strain NM. Strain NM was classified in subgroup (a) by the Millipore filtration and in subgroup (b) by the other two methods.
Bovine Diarrhea Virus
Authors: Tanaka Yoshio , Inaba Yuji , Omori Tuneyoshi , Matumoto Minoru
Published: November 2013

Virus strain No. 12, one of the new isolates from Japanese cattle described previously, was studied for its physicochemical properties. The new isolate was shown to be very small in size by centrifugation and filtration, being filtrable through Millipore filters of 50 mμ pore size. It appears to be an RNA virus as its replication was not inhibited by 5‐iodo‐2′‐deoxyuridine. The virus was readily inactivated by ether and deoxycholate, and partially by trypsin; was labile at pH 3, not stabilized by 1 m MgCl2 at 50 C, was inactivated by ultraviolet, and withstood repeated freeze‐thawing. Further it was readily inactivated at 56 C but more slowly at 37 C, and was stable at lower temperatures. These findings support the identification of the isolated virus as the bovine diarrhea (BD) virus. The properties of BD virus, i.e. size, type of nucleic acid, ether, chloroform and deoxycholate sensitivities, and acid lability, appear to be similar to those of arboviruses. The trypsin sensitivity of BD virus is similar to the B group of arboviruses, which, unlike the A group, sensitive to trypsin. For the classification of BD virus as well as hog cholera virus, which is closely related, further elucidation of properties, fine structure of the virion, etc., is needed.
Isolation of Polyribosome from Tobacco Plants Infected with Tobacco Mosaic Virus
Authors: Kiho Yukio
Published: November 2013

Polyribosomes have been isolated from tobacco leaves. Upon infection with TMV, a new polyribosome has appeared which is specific for infection, and TMV‐antigenic protein is formed on this polyribosome. This new polyribosome has an S‐value of 360–380. From these results, it is suggested that this TMV specific polyribosome is an aggregate of 60–80 ribosomes which is bound by TMV‐RNA as messenger, and that TMV‐RNA is translated polycistronically.
Transfer Agent of Immunity
Authors: Mitsuhashi Susumu , Kurashige Satonori , Kawakami Masaya , Nojima Tokukichi
Published: November 2013

Detection of the cells which contain antibody was accomplished by a method of immune adherence of human erythrocytes to a single cell, termed SCIA (single cell immune adherence) reaction. Peritoneal exudate cells were collected from mice immunized with flagella of either Salmonella enteritidis or S. tennessee. Serologically specific antibody was detectable in some of the peritoneal exudate cells of such mice. An immune ribonucleic acid (RNA) was extracted from the peritoneal exudate cells of mice immunized with salmonella flagella. When mice were injected intraperitoneally with this preparation, serologically specific antibody was found in some of their peritoneal exudate cells by the SCIA method. This preparation was inactivated by treatment with ribonuclease, but was resistant to proteinases, deoxyribonuclease and anti‐flagella antibody, suggesting that this agent is of RNA nature and does not contain antigen or fragment thereof.
Drug Resistance of Staphylococci
Authors: Kasuga Tokuhiko , Mitsuhashi Susumu
Published: November 2013

It was reported that resistance to tetracycline (TC) in some staphylococcal strains was lost irreversibly without loss of resistance to other drugs when cultured at high temperature, and that the determinant for TC resistance exists on a plasmid. According to the transductional analysis of TC resistance in Staphylococcus aureus E169 it was found that the genes which govern resistance to tetracycline and streptomycin are located close together on a single genetic element. When TC resistance in MS146, one of our stock cultures, was transduced to E169S which had lost TC resistance, TC resistance in transductants was found to be unstable at elevated temperatures. By contrast, TC resistance in transductants MS353 TCr, to which TC resistance was transduced from E169, was indicated to be stable even when cultured at high temperature. From these results, it is strongly suggested that instability of TC resistance in E169 is not accounted for by the genetic properties of the genes which govern TC resistance but those of host cell (E169) itself.
Studies on Parainfluenza Virus Infections among Infants and Children in Sendai
Authors: Numazaki Yoshio , Shigeta Shiro , Ishida Nakao
Published: November 2013

A cell line sensitive enough for the recovery of all parainfluenza viruses and free of simian virus contamination frequently occurring in monkey kidney cells was sought. The VERO cell obtained from African monkey kidney was found suitable for the initial isolation of types 1, 2 and 3 parainfluenza viruses, although the cells did not always allow the successive transfer. Mixed cultures of VERO and HEp‐2 cells were also useful in the recovery of various respiratory viruses including parainfluenza viruses. The characteristics of hemagglutinins of parainfluenza viruses were examined, and type 2 parainfluenza and SV5 viruses agglutinated both guinea pig and green monkey erythrocytes at 36 C, whereas types 1 and 3 parainfluenza viruses agglutinated only guinea pig erythrocytes. Thus parainfluenza viruses were divided into two groups by the presence or absence of hemagglutinins for green monkey erythrocytes. Identification of these parainfluenza isolates, employing HI microtechnique was simple and reliable, even with the first passage harvest, when guinea pig erythrocytes were used and the test read at 36 C. Specific standard antisera for these parainfluenza viruses were prepared by immunizing chickens intravenously and bleeding within a short period. These type‐specific antisera were useful for the identification of parainfluenza isolates by HI test.
Immunological Specificity of Soluble Polysaccharides of Candida Albicans
Authors: Fukazawa Yoshimura , Elinov Nikolay P. , Shinoda Takako , Tsuchiya Takeshi
Published: November 2013

Specificity and immunogenicity of soluble polysaccharides of Candida albicans which were extracted with β‐naphthol, boiling water for 1 hr, and with β‐naphthol from cell residue were studied by agglutination and agglutinin absorption method. Soluble polysaccharides produced sufficient agglutinins of IgG nature. A high degree of specificity of soluble polysaccharides was proved, as specific antigen 7 of C. albicans existed in the soluble fractions. Studies on the nature of macromolecules of soluble polysaccharides performed by column chromatography on Sephadex G‐200 and DEAE‐Sephadex, indicated that molecular sizes and yields of neutral and acidic polysaccharides may vary according to methodology used. Soluble polysaccharide extracted with boiling water had a larger molecular size than that obtained with β‐naphthol. Present data on specificity of soluble polysaccharides as well as previous observations on specificity of IgG antibody suggest that IgG produced by immunization with soluble polysaccharide contains specific antibody factor capable of differentiating C. albicans from C. tropicalis.
Breaking up of Vegetative Mycelium of Nocardia and Streptomyces Cultures on Agar Film
Authors: Uesaka Ichiro
Published: November 2013

Cultures of Nocardia and Streptomyces were grown on agar film on glass slides and the mode of breaking up of their vegetative mycelium was studied. It was found that they broke into shorter filaments by two modes: fragmentation and autolysis. There were two types of fragmentation: one is when the divided cells are separated by empty splits and the other is when the cells are not separated but connected to each other, no splits thus being formed. When autolysis occurred at small intervals along the hyphae and then the intervening cytoplasm disappeared, the interstices appeared as empty splits which could not be distinguished from those caused by fragmentation. With the progress of autolysis, parts of the vegetative mycelia became ghost‐like or beaded, which parts usually remained for a long time in Streptomyces cultures but tended to disappear in Nocardia isolates. The splits which had been caused by fragmentation or by autolysis were sometimes filled with newly developed filaments. Although both fragmentation and autolysis were found in Streptomyces as well as Nocardia, the former was more abundantly observed in Nocardia and the latter was predominant in Streptomyces.
Transfer Agent of Immunity
Authors: Kawakami Masaya , Kitamura Kouichi , Mikami Hirokazu , Mitsuhashi Susumu
Published: November 2013

Antibody formation in vitro was studied using erythrocytes (RBC) as antigen and immunocytoadhesion as the technique for detection of antibody‐forming cells. Spleen cells (SPC) of nonimmune mice gained the ability to produce antibody after treatment with ribonucleic acid (RNA) preparation extracted from allogeneic mice immunized with xenogeneic or allogeneic RBC. It was also found that a small proportion of SPC from individual mice of certain strains formed antibody against autologous RBC when the cells were treated in vitro with RNA preparation obtained from the spleen of an allogeneic mouse immunized with RBC of that individual. No converting ability was observed in the RNA preparation from spleen of nonimmune autologous or allogeneic mice. The converting activity of immune RNA preparation was shown to be sensitive to ribonuclease treatment. These evidences exclude the possible contribution of antigen or fragments thereof in the RNA preparation to the induction of antibody formation in RNA recipient cells.
An Attempt to Establish Experimental Dysenteric Bacilli Cystitis
Authors: Awataguchi Shigemi , Kawano Yoshishige , Kojima Akihiro , Sakuma Sadashige
Published: November 2013

It was demonstrated that transurethral inoculation of the freshly isolated strains of Shigella into guinea pigs led to an acute severe dysenteric cystitis, resulting in the death of a majority of them, whereas stock cultures of the same species did not.
Effects of Formycin B on Tobacco Mosaic Virus Multiplication
Authors: Tezuka Nobuo , Hirai Tokuzo
Published: November 2013

Formycin B (FMB), when applied for 5–6 hr immediately after inoculation, inhibited tobacco mosaic virus (TMV) multiplication in tobacco leaf‐discs and local lesion formation on bean leaves. The inhibitory activity of FMB on local lesion formation was reversed by the addition of adenine, adenosine, and adenylic acid of 5 times in molar concentration as much as FMB, but not by hypoxanthine or uracil. FMB‐treatment did not cause significant changes in total RNA contents and in the synthetic rates in TMV‐inoculated and noninoculated tobacco leaf‐discs. However, 32P incorporation rates into TMV and the amount of TMV synthesized in leaf‐discs were remarkably reduced by FMB‐treatment. 32P‐phosphorylated FMB could not be detected either from FMB‐treated tobacco leaf RNA or from TMV‐RNA by means of column chromatography. 3H‐labeled FMB could not be detected in either TMV or in tobacco leaf RNA. The inhibitory action of FMB against TMV multiplication in tobacco leaf‐discs is discussed.
Studies on Swine Enteroviruses
Authors: Kadoi Katuyuki , Kobori Susumu , Morimoto Tomiaki
Published: November 2013

Of 38 new isolates of enterovirus recovered from fecal specimens of Japanese pigs, 14 isolates were found to be strains of a new serotype (represented by strain IPI) distinct from the five serotypes of swine enterovirus hitherto recognized in Japan. The remaining isolates included serotypes Teschen, T80, SF1 and V13. The two different types of cytopathic effect, which have been recently recognized in swine kidney cell cultures, were also observed with our isolates; type I is characterized by a rounding of cells followed by aggregation and destruction of affected cells, and type II by granulation and degradation of cells with poor clumping of affected cells. Of interest is the finding that the type I strains possess a higher heat stability than the type II strains when heated at 50 C for one hr.
Antigenic Determinants of Salmonella for Production of “Clearance” Factor in Mouse Typhoid
Authors: Nakano Masayasu , Saito Kazuhisa , Toyasaki Norituna
Published: November 2013

The “clearance” factor produced in the peritoneal cavity of mice immunized with killed vaccines prepared from Salmonella typhimurium or S. enteritidis was identified as the specific antibodies elicited by the O side chain of the cell wall polysaccharides in the organisms used as immunogens. After immunization of mice with vaccines prepared from virulent Salmonella strains, complement‐dependent antibacterial antibodies in the serum and “clearance” factors in the peritoneal cavity were found to appear coincidentally, to last for more than one year, and to have the same specificity against the virulent bacterial strains. The relationship between the complement‐dependent antibacterial antibodies and “clearance” factor, and the mechanisms of bactericidal action of these antibacterial agents in experimental typhoid were discussed.
Unmasking of Growth of Dermovaccinia Strain Dairen I in L Cells by Acid Treatment of Cells after Virus Adsorption
Authors: Ando Nobuyo , Matumoto Minoru
Published: November 2013

Strain Dairen I (DI) and strain IHD‐T of neurovaccinia virus were readily inactivated in HCl‐citrate buffer solution, pH 2.2 or 2.6, at room temperature, 23 C, reaching about 10−3 or less survival in 2 min and 10−4 or less in 5 min. Treatment of L cells with the buffer solution, pH 2 to 2.5, at room temperature for 5 min after virus adsorption at 37 C effectively reduced the titer of residual active virus, and provided a simple, effective method to unmask the low‐grade replication of strain DI with little deleterious effect on the cells. The method seems useful for the analytical study on the viral replication, particularly the early virus‐cell interactions. The clonal analysis, preliminary in nature, of the viral yield of DI infected L cells provided data suggestive of selection for viruses better adapted to L cells.
Role of Circulating Leukocytes in the Production of Virus‐Inhibiting Factor or Interferon
Authors: Nagano Yasuiti , Maehara Nobutoshi , Nakamura Wataru
Published: November 2013

The relationship between the number of circulating leukocytes and amount of virus‐inhibiting factor (IF) or interferon appearing in blood stream in response to injection of E. coli endotoxin or Newcastle disease virus (NDV) was studied in mice after whole‐body X‐irradiation. Two days after irradiation with 500 R, the leukocyte count fell to 7% of the normal value. Production of IF, however, was not depressed when induced with endotoxin 2 days after irradiation, while NDV induced a slightly lower titer of IF, 1/1.6 of that in non‐irradiated mice. When induced 5 days after irradiation, though leukocyte count remained at 8 to 9% of the normal value, IF titers attained were only 1/3 and 1/4 of the control value with endotoxin and NDV, respectively. In mice given 1000 R before 2 days, leukocyte number was less than 1% of the normal value, while IF titers reached 1/2.7 and 1/5 of that in the control mice with endotoxin and NDV, respectively. The lack of parallel between the leukocyte count and amount of IF produced suggest that circulating leukocytes do not produce IF.
Lability of Streptomycin Resistance in Staphylococcus aureus
Authors: Morimura Masato , Watanabe Ken , Mori Hiromichi , Mitsuhashi Susumu
Published: November 2013

The determinant for streptomycin resistance in some strains of Staphylococcus aureus isolated from clinical specimens was found to be lost during storage as stock cultures or by high temperature treatment. This fact has added an additional proof that many genetic determinants for drug‐resistance in staphylococci are easily and irreversibly lost.
Heat‐Labile Virus‐Inhibiting Factor or Interferon Induced by Virus
Authors: Maehara Nobutoshi , Nagano Yasuiti
Published: November 2013

Stability to heat and acid of the virus‐inhibiting factor (IF) or interferon induced by viruses is generally accepted. Published data suggest, however, a heterogeneity in the virus‐induced IFs in their physico‐chemical and biological properties. The present work shows that the IF activity of rabbit serum collected 2 hr after intravenous inoculation with Newcastle disease virus is as labile as E. coli endotoxin‐induced IF when heated at 56C or dialysed at pH 2, whereas the serum‐IF at 6 hr is significantly more stable.
Relationship between Immunity and Interferon Production in Macrophages
Authors: Yamada Morihide , Azuma Masanobu , Nishioka Rikichi , Togashi Takehiro
Published: November 2013

In vitro interferon (IF) production in peritoneal macrophages of normal and Newcastle disease virus (NDV)‐immunized mice was studied. Of ascites cells used, 80% were macrophages, 14% lymphocytes, and 6% polymorphonuclear leukocytes. It was indicated that IF was produced mainly in the macrophages after NDV inoculation. IF production in the macrophages derived from immunized mice was more enhanced than that in those from normal mice. It is not clear at present, however, whether this enhancement is based on immunological specificity. The IF production in the culture of macrophages reached its maximum value in 6 to 9 hr after inoculation of the inducer. After 12 hr, the IF titer in the culture fluid decreased gradually. A possible explanation of this fact is that there may be partial inactivation of IF by some cellular components.
Relationship between Immunity and Interferon Production in Macrophages
Authors: Azuma Masanobu , Yamada Morihide , Nishioka Rikichi
Published: November 2013

When Newcastle disease virus (NDV) was inoculated into macrophage cultures, interferon (IF) of various molecular weights appeared in the culture fluid. The molecular weight of IF produced in the macrophages derived from normal mice in at least 6 hr post‐inoculation of inducer was different from that produced in the macrophages from immunized mice. The IF with a molecular weight as high as 100 000 was unstable against pH 2 treatment and heat treatment, while that of 70 000 or less was stable.
Intestinal Resistance in the Experimental Enteric Infection of Mice with a Mouse Adenovirus
Authors: Hashimoto Kazuo , Sugiyama Takiko , Yoshikawa Masako , Sasaki Shogo
Published: November 2013

Investigation was made on the process of enteric infection with mouse adenovirus strain K87 in inbred DK1 mice and the intestinal resistance acquired through infection. The cells containing viral antigens were enumerated in most parts of the infected intestinal tract by a fluorescent antibody technique, and the infectivity titer of the virus in each part was examined in mouse kidney tissue culture. The virus was observed to grow in 3~14 days (sometimos 3~21 days) after oral challenge, and infectivity titers reached their peak after 7~14 days, when a number of viral antigen‐containing cells and cells with nuclear inclusions were detected. In the mice rechallenged 28 days after the initial challenge, the virus did not grow, and no viral antigen‐containing cells were found. From these results it was concluded that the main sites where the virus grows in mice are the cells which are scattered in the epithelial layer of the mucous membrane of the small intestine, and which seem to be the usual epithelial cells and not Paneth's or goblet cells. As for intestinal resistance, experiments with inactivated vaccine and with passive transfer of serum‐antibodies were performed in order to find out whether neutralizing antibodies in the serum had any influence on the growth of virus in the intestinal wall, and no influences were indicated. Eighteen days or more after challenge, K87 virus‐neutralizing substances were detected in the intestinal wall and in the intestinal contents of the infected mice, but not in the serum‐transferred mice, though both groups of mice had equal levels of serum antibodies. The substance continued to be found until 15 weeks after challenge in the intestinal contents, and until later than 34 weeks in the intestinal walls. The nature and the possible role of the substance is discussed, but actual data will be reported in subsequent papers.
Transduction of R Factors with Phage P1 from Recombination‐Deficient Escherichia coli
Authors: Hashimoto Hajime , Iyobe Shizuko , Mitsuhashi Susumu
Published: November 2013

In the transduction of R factor with phage P1 from rec− host, transduction frequency was less than that from the rec+ host but no segregation of resistance markers took place. However, a low percent of segregation was found in the transduction from rec+ host. In contrast, phage particles of small size contained in P1 lysate could not transduce R factor even from a rec+ host, although they could transduce chromosomal markers as did the usual P1 lysate. Recombination of the R factor took place in the rec− host at a frequency similar to the rec− host. Considering from the results that P1 is capable of transducing a DNA fragment of the same size as that of phage P1, it was concluded that the molecular weight of R factor DNA is smaller than that of phage P1 (6×107 daltons) but larger than that of small P1 particle (2.4×107 daltons). When transduced with phage P1, the R factor genome has to become a DNA fragment of the same size as the P1 particle by addition of other DNA following the recombination between R factor and Pl in rec+ host. Hence, the segregation of markers of the R factor takes place on occasion by this recombination. From the assumption that phage Pl cannot recombine with the R factor in rec− host, it was predicted that the R factor genome becomes a DNA fragment of large enough size for transduction with P1 by polymerization of the R genome itself. This possibly could explain the decrease in transduction frequency from a rec− host and no segregation of the resistance markers of R factor in transduction from rec− host.
Behavior of Bovine Ephemeral Fever Virus in Laboratory Animals and Cell Cultures
Authors: Matumoto Minoru , Inaba Yuji , Tanaka Yoshio , Ito Hiroshi , Omori Tuneyoshi
Published: November 2013

A Japanese strain (Yamaguchi) of bovine ephemeral fever virus, propagated in BHK2I‐WI2 cells of hamster kidney origin, was used. Adult rats inoculated intracardially or intracerebrally remained healthy, but produced neutralizing antibody, suggesting actual viral growth in the animals. However, no clinical and serological evidence for infection with the virus was obtained in adult sheep, goats, swine and rabbits inoculated intravenously. One‐day‐old guinea pigs inoculated intracerebrally and adult guinea pigs inoculated intracardially remained healthy and produced no neutralizing antibody. The virus multiplied not only in BHK2I‐WI2 cell culture but also in cultures of stable line hamster cells (HmLu‐1 and HmT), primary hamster kidney and embryo cells, long‐term cultured bovine embryo cells (BEK1), primary calf kidney and embryo cells, and primary suckling rat kidney and embryo cells. However, no evidence for cytopathogenesis and viral growth was obtained in primary cultures of bovine leukocytes, bovine bone marrow. thymus, thyroid and adrenal cells, guinea pig, mouse and chick kidney cells, and mouse and chick embryo cells, and in culture of African green monkey kidney cells (Vero). The virus multiplied best and induced marked cytopathogenesis in BHK21‐WI2 cells; lower temperatures such as 34 C or 30 C appeared preferable to obtain constantly better viral yields, although the virus grew more slowly. Similar effects of the incubation temperatures were also obvious in the other susceptible cell cultures, in which the cytopathogenesis was less marked than in BHK21‐WI2 cells or absent, roughly coinciding with decreasing grades of viral growth.
Absence of an All‐or‐None Type of Complement Requirement in Early Neutralizing Antibody against Western Equine Encephalitis Virus
Authors: Yoshino Kamesaburo , Morishima Taka , Aoki Yoshiro
Published: November 2013

Rabbits were immunized by 2 weekly intravenous inoculations of beta‐propiolactone‐inactivated Western equine encephalitis virus vaccine followed 1 week later by a subcutaneous inoculation of live virus, and complement requirements of early and late neutralizing antibodies were compared. There was no difference between these antibodies, both showing about a 4‐fold enhancement of the virus‐neutralizing activity in the presence of complement. This was also the case when guinea pigs were given the vaccine by the intraperitoneal route. It is unlikely that previous exposure of the animals to an antigenically related virus caused a booster effect upon the antibody development, because the rabbit antibody could not even neutralize the closely related Sindbis virus either in the presence or in the absence of complement. The reason for the existence of 2 different types of primary responses with respect to the complement requirement of early neutralizing antibody is discussed.
Induction of Interference and Interferon Production by Echovirus Type 7
Authors: Hoshino Masao
Published: November 2013

Echovirus type 7 does not multiply in mice, but injection of this virus into mice (intracerebrally or intraperitoneally) increased their resistance to a subsequent infection with Coxsackie B virus, type 5 and to Japanese encephalitis virus by intracerebral route. The effect is evident in the survival rate and the prolongation of life among the mice that did succumb to Coxsackie or Japanese encephalitis virus infection, as well as those that developed paralysis. An adequate dose of active interfering virus and a time interval between the two inoculations were both necessary to obtain a significant interfering effect. Pretreatment of mice with actinomycin D suppressed the interfering effect by Echovirus type 7, and the kinetics of interferon production corresponded to the time of establishment of interference. All these findings suggest that the induction of resistance by Echovirus type 7 is mediated by interferon produced in the early stages of infection.
High Incidence of Adults with Rubella Antibody in Northern Japan
Authors: Suto Tsunehisa , Morita Morihiro , Hinuma Yorio , Ishida Nakao
Published: November 2013

A seroepidemiological study of rubella in northern Japan was made by testing for hemagglutinin‐inhibitory (HAI) antibodies to rubella virus using an HAI test with our modification. An epidemic of exanthematous disease, which occurred in 1966–1967, was confirmed as rubella by the age specific distribution of rubella HAI antibody. However, one of the 14 areas tested in the region, a Piedmont village, was found to have had no rubella experience during the last 15 years. In general, a higher incidence of high titered HAI antibody was observed among the younger ages, suggesting a decrease of antibody titer with an increase in age after infection. Through these studies, it is evident that 100% of adults over 20 years of age possessed HAI antibody in all age groups and all areas so far examined. This fact may possibly explain the extremely rare incidence of congenital rubella syndrome in the northern part of Japan.
Cellular Mechanisms of Adjuvant Action of Bacterial Lipopolysaccharide in Anti‐Sheep Red Blood Cell Antibody Response
Authors: Nakano Masayasu , Shimamura Tadakatsu , Saito Kazuhisa
Published: November 2013

Adjuvant action of lipopolysaccharide (LPS) extracted from Salmonella typhimurium LT2 on anti‐sheep red blood cell (sRBC) antibody response in mouse spleen was studied at the cellular level by the technique of localized hemolysis in agar. Injection of LPS caused significant increase in direct 19S and indirect 7S plaque‐forming cells (PFC) in mouse spleen after the primary antigenic stimulation, while the adjuvant effect of LPS could hardly be assessed in the secondary PFC response. The adjuvant effect on the primary PFC response was most conspicuous when LPS was injected at the same time as the sRBC, and no effect could be observed when LPS was injected 1 or 5 hr before PFC determination. The mice who had received LPS with the primary sRBC stimulation showed much higher PFC level after the secondary sRBC stimulation, indicating the increase in the memory cells. Cell population consisting of lymphoid cells and macrophages was separated into ‘adherent’ (macrophage‐rich) and ‘non‐adherent’ (lymphoid cell‐rich) fractions by incubating in petri dishes, and effect of LPS on each fraction was examined. Transfer of non‐adherent lymphoid cells treated with LPS in vitro and simultaneous sRBC injection resulted in a significant increase of PFC in the spleen of the syngeneic recipient mice, but transfer of LPS‐treated macrophages did not show any effect, suggesting that LPS acts on lymphocytes but not on macrophages. This idea appeared also to be supported by the results that LPS exerted antagonistic effect on the immune suppression caused by heterologous anti‐lymphocyte serum, anti‐sRBC serum and cortisone which are currently supposed to suppress the antigen reactive cells.
Phosphorylated Inactivation of Aminoglycosidic Antibiotics by Pseudomonas aeruginosa
Authors: Kobayashi Fujio , Yamaguchi Masahito , Mitsuhashi Susumu
Published: November 2013

The studies were conducted on the inactivation of aminoglycosidic antibiotics including a new antibiotic, lividomycin, by a cell‐free fraction from a strain of Pseudomonas aeruginosa isolated from the urine of a patient with cystitis. The strain strongly inactivated No. 2230‐C (mannosyl paromomycin), paromomycin, kanamycin, aminodeoxykanamycin and neomycin but did not inactivate lividomycin A, dihydrostreptomycin and gentamicin. Inactivated‐No. 2230‐C, ‐paromomycin and ‐kanamycin were isolated and purified using column chromatography techniques. Inactivated antibiotics were found to be reactivated by the treatment with alkaline phosphatase. According to the reactivation by alkaline phosphatase and analytical data, it was concluded that the inactivated products were monophosphorylated derivatives of each antibiotic.
Studies on the Persistent Infection with Measles Virus in HeLa Cells
Authors: Minagawa Tomonori , Yamada Morihide
Published: November 2013

Almost all cells in HeLa/MV culture contained measles antigen which was mainly detected on the cell surface by immunofluorescent and hemadsorption technique. Cytotoxic effect on HeLa/MV cells was demonstrated by anti‐measles sera and complement (immunolysis), indicating the existence of viral surface antigen. Anti‐measles serum absorbed with purified measles particles completely lost the ability of immunolysis. Though the cytolysis by anti‐measles sera was due to antigen‐antibody reaction on the cell surface, the mechanism of lysis seemed to be different from that of the cytolysis by anti‐HeLa cell serum. It was suggested that some unknown serum factors other than components of complement are necessary for immunolysis by anti‐measles serum.
Recombination between Mycobacterium smegmatis Strains Jucho and Lacticola
Authors: Mizuguchi Yasuo , Tokunaga Tohru
Published: November 2013

Genetic recombination was demonstrated in the genus Mycobacterium. Auxotrophic mutants of M. smegmatis strain Jucho requiring different growth factors did not yield prototrophic recombinants when progenies from mixed cultures were inoculated onto minimal medium. Similarly, mutants of M. smegmatis strain Lacticola did not produce prototrophic recombinants. When mating was attempted between auxotrophs of the strains Jucho and Lacticola, prototrophic recombinants were recovered at frequencies of about 10−3 to 10−5 of total cells, while the frequency of reversion of either parent was less than 10−7. The segregation of unselected characters was noticed in recombinants resulting from interstrain matings. No prototrophic recombinant was detected in any cross in liquid media. Incubation of mutants of one strain with a culture filtrate or deoxyribonucleic acid of another compatible strain produced no recombinant. It was suggested that a conjugal mechanism rather than a transforming or transducing principle played an important role in the above observed gene‐transfer. Pretreatment of parental cells with acridine dyes did not eliminate the ability to produce recombinants.
Genetical Distinction of R Factors Derived from Shigellae and Salmonellae
Authors: Yoshikawa Masanosuke , Nagashima Shigeaki , Matsushima Shingo
Published: November 2013

Comparison of R factors derived from shigellae and salmonellae was made using several genetical characteristics. Almost all R factors derived from shigellae were fi+ (fertility inhibition), spp− (no restriction against phage λ vir) and possessed a drug resistance pattern of (sul, str, cml, tet), whereas those from salmonellae were fi−, spp− and their resistance patterns were (str), (tet) or (str, tet). The (str, tet) R factors often segregated upon conjugal transfer but R factors from shigellae did not. R factors from shigellae were unstable in Salmonella but stable in Shigella and Escherichia coli. R factors derived from salmonellae were unstable in Shigella but stable in Salmonella and E. coli. All fi+, spp− R factors derived from shigellae, except two which had been known to be exceptional also in the incompatibility tests, conferred atabrine sensitivity (ats+) upon the host cells, whereas all fi−, spp− R factors derived from shigellae were ats−. One exceptional fi−, spp− R factor derived from Shigella was ats−, but all of the fi−, spp− R factors derived from salmonellae were ats+. Two representative R factors derived from salmonellae were shown to be compatible with 14 fi+ and fi− standard R factors which were derived from shigellae and had been stocked in 3 laboratories in Japan as representative collections. Five (tet) segregants originally derived from (sul, str, cml, tet) R factors of shigellae but obtained by transferring to and segregating in Salmonella panama were shown to be different from (tet) R factors of Salmonella origin with respect to the fi character and the incompatibility tests. From these results it was concluded that R factors of Shigella origin and those of Salmonella are different.
Aspartic Acid Transport in Mycobacterium smegmatis
Authors: Yabu Kunihiko
Published: November 2013

L‐Aspartic acid was taken up by Mycobacterium smegmatis cells at a greater rate than D‐aspartic acid. Kinetic studies of L‐aspartic acid uptake revealed a curvilinear Lineweaver‐Burk plot. The observed kinetics could be accounted for by the existence of two different processes, a saturable and a nonsaturable. In contrast, the latter process was solely active for D‐aspartic acid uptake. Competition studies indicated that aspartic acid and glutamic acid were similarly affected by the operation of 2 different processes. The existence of 2 similar catalytic transport processes, specific for dicarboxylic amino acids, was discussed.
Responses of Calf Kidney Cells to Human Adenovirus Type 12
Authors: Kimura Susumu
Published: November 2013

The process of infection of established calf kidney (CKT) cells with human adenovirus type 12 (Ad 12) was compared with that with type 5 (Ad 5). Ad 5 replicated in CKT cells but Ad 12 did not. In CKT cells infected with Ad 5, as much virion antigen was detected by complement‐fixation (CF) test as from HeLa cells, and the rate of DNA synthesis increased to about 7 times that in uninfected cells. In CKT cells infected with Ad 12, production of T‐antigen was detected by CF test, but neither production of virion antigen nor increase in DNA synthesis was observed. Confluent monolayers of CKT cells incorporated 3H‐thymidine progressively into DNA after growth medium was replaced with maintenance medium. This increase in host cell DNA synthesis after changing media was inhibited by infection with Ad 12 or with ultraviolet‐irradiated Ad 12.
Interferon‐Inducing Activity of Acidic Polysaccharides
Authors: Suzuki Shigeo , Suzuki Masuko , Imaya Masanori
Published: November 2013

Exocellular phosphomannans produced by four strains of Hansenula yeast were examined as to their interferon‐inducing activities in rabbits employing the assay system consisting of vesicular stomatitis virus and primary suckling rabbit kidney cells. The phosphomannan of Hansenula holstii NRRL Y‐2155 was shown to be the most potent inducer, and the interferon induced was highly host species‐specific, preventing viral cytopathic effects only in the homologous cells. The biological and chemical characteristics of the interferon closely resembled those of endotoxin‐induced interferon.
Delayed Type Hypersensitivity in Hamsters Infected with Mycoplasma pneumoniae as Revealed by Macrophage Migration Inhibition Test
Authors: Arai Sumio , Hinuma Yorio , Matsumoto Keizo , Nakamura Takashi
Published: November 2013

A delayed type hypersensitivity was demonstrated in hamsters infected with a fresh isolate of Mycoplasma pneumoniae by the following two procedures of macrophage migration inhibition test. In the first procedure, inhibition of migration of peritoneal exudate cells from normal guinea pigs by the addition of splenic cells from infected hamsters in the presence of the mycoplasma antigen was examined. In the second procedure, migration of peritoneal exudate cells, composed of macrophages and lymphocytes, from infected hamsters was examined after the addition of the antigen. Virtually, both procedures gave a similar result, suggesting the presence of cell‐mediated immunity in the infected animals. In these infected hamsters, humoral antibody responses were also demonstrated at the same time.
Double Strandedness of Ribonucleic Acid of Bovine Ephemeral Fever Virus
Authors: Tanaka Yoshio , Inaba Yuji , Ito Yasuichiro , Sato Kunihiko , Omori Tsuneyoshi , Matumoto Minoru
Published: November 2013

Virus labeled with 3H‐uridine or 32P‐orthophosphate was purified by CsCl equilibrium centrifugation of concentrated virus materials from infectious BHK21‐WI2 cell culture fluids. A single clearly visible band formed in the gradient coinciding with a sharp peak of radioactivity having a buoyant density of 1.19 g/ml. Infectivity exhibited a broader distribution with a peak coinciding with the visible band, in which numerous virions of the virus were observed with the aid of an electron microscope using the phosphotungstic negative staining technique. Ribonucleic acid (RNA) extracted from the purified virus by the phenol method exhibited a rather broad distribution of radioactivity with a major peak at about 12 S when analyzed by the sucrose density gradient centrifugation technique. Viral RNA centrifuged in the gradient after ribonuclcase (RNase) treatment showed a single sharp peak at about 12 S. These findings seemed to indicate that the virion of this virus contained double‐stranded RNA. Double strandedness of the viral RNA was further corroborated by an examination of the base composition, reduced resistance to RNase in low salt concentration and a sharp thermal transition with a relatively high melting temperature of 85 C. The virus could not be classified either in the rhabdovirus group, although the shape of the virion resembled that of the rhabdoviruses, or in the reovirus group since the virus was ether‐sensitive. It seemed necessary to create a new genus for this virus.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Kuroiwa Ataru , Takeya Kenji
Published: November 2013

In a previous paper we reported that an inbred strain of SL mice and an outbred strain of CF1 mice belonged to the high‐responder strains in antibody production after primary immunization with hamster erythrocytes (H‐RBC), while inbred strains of C57BL/6, AKR and C3H/He mice belonged to low‐responder strains. In the present study we obtained the following results. 1) Pre‐sensitization with hamster lymphoma enhanced antibody production after an intravenous injection of H‐RBC. There was no strain difference in the pattern of antibody production against H‐RBC among pre‐sensitized mice. 2) The pattern of enhanced antibody production after an intravenous injection of H‐RBC into pre‐sensitized mice assumed the primary type in terms of time of appearance of hemolysin plaque‐forming cells (PFC) in the spleens and the conversion from 2‐mercaptoethanol sensitive to 2‐mercaptoethanol resistant antibody production, when the intervals between both treatments were within 7 days. 3) Pre‐sensitization with lymphoma induced not only an increase in numbers of PFC after an intravenous injection of H‐RBC, but also an increase in the size of the hemolysin plaques. These results suggested that sensitization with hamster lymphoma stimulated some kinds of immuno‐competent cells, which could contribute to antibody production against H‐RBC after a booster injection of H‐RBC.
Studies on the Neutralization of Herpes Simplex Virus
Authors: Yoshino Kamesaburo , Morishima Taka
Published: November 2013

A portion of the seed virus remained active even in the presence of excess antibody. With an early serum or relatively low concentrations of a hyperimmune serum, the presence of such unneutralizable virus was uncovered by the addition of complement. This unneutralizable persistent fraction (PF) was distinguished from sensitized virus surviving in insufficient antibody, because the latter was inactivated by a high concentration of antibody as quickly as the control virus. The level of the PF was constant regardless of the serum species, serum lot, dilution of serum, antibody class and complement requirement of antibody, being approximately 0.1% of the seed virus. Millipore filtration of the seed virus lowered this PF level to varying degrees depending upon the porosity. However, when filtered virus‐serum mixtures were incubated at 37 C for a sufficient time and then filtered again, a portion of the unneutralized virus did pass a 0.22 μ membrane. Furthermore, virus sensitized with insufficient antibody showed no increase in resistance to complement after 10 hr incubation at 4 C. These results proved that viral aggregation was not the essential cause of the PF. It was confirmed, on the other hand, that the neutralization of virus by hyperimmune IgG followed a one‐hit curve. Analysis of these data appeared to support Rappaport's postulation that the PF represented antibody‐coated virus whose critical antigenic sites were all left unbound.
Characterization of the Chromosomally Mediated Penicillinase in Klebsiella pneumoniae
Authors: Matsumoto Hideki , Sawai Tetsuo , Tazaki Tadakatu , Yamagishi Saburo , Mitsuhashi Susumu
Published: November 2013

The gene governing ampicillin resistance and penicillinase production in Klebsiella pneumoniae was analyzed by means of a conjugation system. The gene, amp, was mapped between pyr4 and aro18 on the linkage map of K. pneumoniae. The chromosomally mediated penicillinases of k. pneumoniae were partially purified and their properties were compared with those of known species‐specific penicillinases of K. pneumoniae and also those of the penicillinases of R factors having ampicillin resistant marker. The chromosomal penicillinases were consistent with most of the species‐specific penicillinases in enzymological and immunological properties and in their behavior on agar gel electrophoresis. They also showed a close relationship to type I penicillinases of R factors in enzymological and immunological properties.
Formation of Multiple Drug Resistance Factors by Recombination between Transfer Factor and Resistance Determinants
Authors: Kameda Mitsuo , Harada Kenji , Suzuki Mitsue , Nakajima Takashi , Mitsuhashi Susumu
Published: November 2013

Drug‐resistance(r) determinants, r21(tet), r1(cml), and r11(str.sul), were obtained from R factors, and were capable of conferring resistance to tetracycline, chloramphenicol, and to both streptomycin and sulfanilamide, respectively. They were integrated into the chromosome of Escherichia coli K12 and nontransmissible by conjugation. As described previously, the recombinant T‐tet factor was formed and conjugally transmissible when E. coli K12 r21(tet)+ was infected with T95, one of the transfer factors. Similarly, the recombinants T‐tet.cml and T‐tet.str.sul factors were formed by the interaction between T‐tet factor and resistance determinants r1 (cml) and r11(str.sul), respectively. Subsequently, T‐tet.cml.str.sul factor was formed by the recombination between T‐tet.cml factor and r11(str.sul) determinant, and T‐tet.str.sul.cml factor was produced by the infection of E. coli K12 r1(cml)+ with T‐tet.str.sul factor. These recombinants were conjugally transmissible and transduced by phage Pl as one unit. These results strongly suggested the origin of R factors which were capable of conferring multiple resistance and transmissible by conjugation. According to the segregational patterns of resistance determinants by transduction and by conjugal transmission, the genetic structure of T‐tet.cml.str.sul factors was established as being circular and the linkage order of the determinants of these factors was described.