bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunochemical Studies on Bacterial Blood Group Substances
Authors: Furukawa Ken , Kochibe Naohisa , Takizawa Hisao , Iseki Shoei
Published: November 2013

A‐decomposing enzyme preparation (α‐N‐acetylgalactosaminidase) from hog liver, liberates from the lipopolysaccharide of Salmonella riogrande, which contains O antigen 40, 15.5% of its original amount of N‐acetylgalactosamine, destroying its ability of inhibiting the agglutination of human A red ceils by anti‐S. riogrande rabbit serum. From the structures of blood group A‐active oligosaccharides, obtained from the lipopolysaccharide by mild acid‐hydrolysis, it was assumed that the antigenic determinant of A specificity might have the following structure, which indicated the presence of N‐acetylgalactosaminyl residue as the non‐reducing end; α‐N‐acetylgalactosaminyl‐(1→3)‐mannosyl‐glucosyl‐(1→3)‐N‐acetylgalactosaminyl‐glucose.
Studies on Bacteriophage Distribution
Authors: Dhillon T. S. , Dhillon K. S. Elvera
Published: November 2013

Twenty‐eight bacteriophage strains active on one or more species of Enterobacteriaceae were isolated from sewage and were classified into seven host range groups. Two isolates resemble S13, three isolates appear to be closely related to T1, and one seems to be related to T even phages. Members of one host range group are active on three bacterial species, viz., Escherichia coli, Salmonella typhimurium, and Shigella sonnei. Some of the E. coli K12 strains selected for resistance to phage HK019 are sensitive to S13 but not to φ×174.
Electron Microscopic Studies on the Outer Layers of Staphylococcus aureus Using a Lytic Enzyme from Flavobacterium
Authors: Futai Masamitsu , Okabayashi Kinji , Mizuno Den'ichi
Published: November 2013

The structure of the outer layers (cell wall and membrane) of Staphylococcus aureus was studied by electron microscope using a bacteriolytic enzyme from Flavobacterium sp. called the L‐11 enzyme. Comparative studies on the morphology of bacteria before and after treatment with this enzyme and cell wall and membrane fractions obtained from bacteria after the enzyme treatment led to the following conclusions. (1) The cell wall of S. aureus is composed of morphologically distinct two layers which are both susceptible to the L‐11 enzyme. (2) Between the cell wall and membrane, there is an electron opaque region which could not be stained using any of the methods tested. (3) Before treatment of bacteria with the enzyme the cell membrane could not be seen clearly. However, after enzyme treatment the membrane was clearly seen. (4) The infolding of the inner layer of the cell wall, forming a structure like a mesosome, was liberated by extensive enzyme treatment.
Antagonism of Non‐Inducer Ribonucleic Acid against Induction of Antiviral Cell Resistance and Interferon by Inducer Ribonucleic Acid
Authors: Kawade Yoshimi , Matsuyama Masako , Yamamoto Yoko , Ozaki Hiroshi
Published: November 2013

Various single‐stranded RNAs, which do not induce antiviral cell resistance in L and chick embryo cells, were found to antagonize the action of inducer RNAs such as poly(C)·poly(I). The non‐inducing RNA inhibited production of interferon stimulated by inducer RNA, but did not inhibit the action of exogenously added interferon. Two aspects were discerned in the antagonism of non‐inducing RNA, namely intrinsic antagonism and anti‐protamine effect. The intrinsic antagonism is the one observed in the absence of protamine. The anti‐protamine effect is elimination of the potentiating action of protamine on inducing RNA [10] by non‐inducing RNA in a large excess over protamine. The relative importance of the two effects will vary depending on the concentrations of protamine and non‐inducing RNA used. Non‐inducer RNA added to cells pretreated with inducer RNA for 1–3 hr effectively inhibited the development of antiviral cell resistance, but was not very effective when added prior to inducer RNA. RNA from MS2 phage virions was found to be an inducer in chick embryo cell but a non‐inducer and antagonist in human embryo lung fibroblasts, indicating that a given RNA may possess both the inducer and the antagonist characters. The induction of interferon by viruses was not significantly inhibited by non‐inducing RNA. The results obtained are consistent with an assumption that non‐inducing RNA competitively inhibits the binding of inducer RNA to cellular receptor sites, although alternative interpretations are not excluded.
Properties of R Factors Isolated from Multiple‐Resistance Proteus rettgeri
Authors: Odakura Yoshihiro , Tanaka Tokumitsu , Yumoto Makoto , Inafuku Zensho , Mitsuhashi Susumu
Published: November 2013

Majority (80%) of the 40 strains of Proteus rettgeri isolated from clinical specimens were resistant to seven drugs: [chloramphenicol (CM), tetracycline (TC), dihydrostreptomycin (SM), sulfanilamide (SA), kanamycin (KM), ampicillin (APC), and cephaloridine (CER)], followed by those resistant to six drugs (15%), single TC‐resistance (2.5%) and non‐resistant strains (2.5%), in decreasing orders. Ninety‐five percent of the 39 multiple drug‐resistant strains conjugally transferred some of their resistance markers and 92% of the R+ P. rettgeri were found to be in a hetero‐R state. The R factor conferring single KM‐resistance was isolated most frequently but TC resistance was never transferable. Detailed analysis of two strains of P. rettgeri in a hetero‐R state, GN3162 and GN3163, using two recipients, Escherichia coli ML1410 and P. rettgeri GN3195, disclosed that resistance patterns of conjugants obtained from the same donor strain were almost identical in both recipients of the different species. When strain ML1410, which had received the whole pattern of multiple resistance of the original isolate, was used as a secondary donor, separate transfer of R factors was again observed. The component of R factors in the hetero‐R strains was determined by transduction experiments as R (KM), R (APC), R (CM. KM) and R (SM. SA). All of these R factors were fi–. Reconstruction of the original hetero‐R state was carried out by superinfection either in E. coli or P. rettgeri, and one of the original strains was observed to be carrying three kinds of R factors, viz., (CM. KM), (SM. SA), and (KM).
Origin of Thymidine Kinase in the Cells Infected with Infectious Canine Hepatitis Virus
Authors: Shinagawa Morikazu , Yanagawa Ryo
Published: November 2013

Thymidine kinase (TK) was induced in dog kidney cells (DKC) and hamster embryo cells (HEC) infected with infectious canine hepatitis virus (ICHV). The enzyme activity increased and reached levels of 13 and 19 times as much as in uninfected cells by the 30th hr after infection in virus‐infected DKC and HEC, respectively. No difference in the pattern of inactivation at 45 C was found between the TK of infected and uninfected cells. The activity of the TK from ICHV‐infected DKC was not inhibited by a dog serum hyperimmunized against ICHV‐infected DKC. From these results it was concluded that the TK which increased in ICHV‐infected cells was of the cellular origin.
Slime Production by Pseudomonas aeruginosa
Authors: Murakawa Takeo
Published: November 2013

Two samples of slime obtained from Pseudomonas aeruginosa strains, IFO 3445 and No. 24, the latter which produced mucoid colonies on brain heart infusion agar as well as on the synthetic agar medium, were investigated for their physicochemical properties, primarily for their viscosities. Results obtained indicated that the principal component of the slime from strain IFO 3445 might be a deoxyribonucleic acid‐like substance, while the slime from the mucoid strain No. 24 might be an alginic acid‐like substance.
Distribution of R Factors among Shigella Strains Isolated in Japan (II)
Authors: Tanaka Tokumitsu , Tsunoda Mitsuko , Mitsuhashi Susumu
Published: November 2013

Shigella strains isolated in Japan from 1968 through 1970 were surveyed for drug resistance and distribution of R factors. Of the 2688 strains, 93.4% were resistant to either one or various combinations of four drugs, tetracycline (TC), chloramphenicol (CM), streptomycin (SM) and sulfanilamide (SA). Among these resistant strains, 74.2, 10.7, 1.48, and 13.6% were quadruply, triply, doubly, and singly resistant, respectively. Fifty‐eight per cent of these resistant strains were found to carry R factors when judged by transferability of the resistance. The isolation frequencies of R (TC. CM. SM. SA), R (CM. SM. SA), R (SM. SA), and R (TC. SM. SA) factors were 73.2, 13.0, 11.5, and 1.3%, respectively. The strains resistant to drugs other than the aforementioned four were very few; 4.3, 3.4, and 0.7% being resistant to ampicillin (APC), nalidixic acid (NA), and kanamycin (KM), respectively. Among 117 APC‐resistant strains, 97.4% could transfer their APC resistance together with other resistance markers. Seventeen out of 18 KM‐resistant strains could transfer KM resistance by mixed culture. But none of the NA‐resistant strains could transfer their NA resistance. The authors could demonstrate strains carrying two different R factors in a cell and one of them was consistently an R (SM. SA) factor. These results were very similar to those obtained in surveys of strains isolated from 1965 through 1967.
Intracellular Components Associated with Chikungunya Virus‐Specific Ribonucleic Acids in Infected BHK21 Cells1
Authors: Igarashi Akira , Sasao Fuyoko , Fukai Konosuke
Published: November 2013

Cytoplasmic extracts of BHK21 cells infected with Chikungunya virus were analyzed by sucrose gradient sedimentation. Besides 140 S nucleocapsid, 65 S component associated with 26 S single‐stranded ribonucleic acid was labeled with 3H‐uridine in actinomycin‐treated infected cells. The 65 S component, demonstrable in ethylenediaminetetraacetic acid‐containing buffers, appeared to accumulate when protein synthesis was inhibited. After glutaraldehyde fixation, this component had a density of 1.48–1.50 g/ml in CsCl. Pulse‐chase experiments did not indicate that the 65 S component was a precursor of the 140 S nucleocapsid.
Replication of Adenovirus 12 Deoxyribonucleic Acid in Association with the Nuclear Membrane
Authors: Yamashita Tadashi , Shimojo Hiroto
Published: November 2013

Analysis of nuclei of adenovirus 12‐infected cells revealed that viral DNA replicated in association with the nuclear membrane and that complete viral DNA was liberated from the nuclear membrane. Analysis of isolated nuclei in vitro showed that DNA polymerase activity increased in the nuclear membrane of adenovirus 12‐infected cells without addition of primer DNA.
The Role of Complement in Immune Phagocytosis of Group A Streptococci by Mouse Peritoneal Phagocytic Cells
Authors: Saito Yuichi , Horikawa Takahiro , Kondo Wataru
Published: November 2013

The role of complement in ingestion of nonencapsulated virulent group A streptococci by mouse peritoneal phagocytic cells was studied in vitro in the presence of type‐specific IgG or IgM anti‐M antibody. Type‐specific IgG anti‐M antibody significantly enhanced the phagocytosis, whereas IgM antibody was less effective in opsonization of the virulent group A streptococci. The extent of phagocytosis of streptococci sensitized with IgG or IgM anti‐M antibody was not influenced by the presence of complement. These results suggested that complement was not an essential factor for efficient phagocytosis of virulent group A streptococci.
Effect of Dimethyl Sulfoxide on Natural Resistance to Bacterial Infections in Mice
Authors: Kato Nobuo , Ohta Fujito , Kato Osamu , Kobayashi Takashi , Nakashima Izumi
Published: November 2013

Dimethyl sulfoxide (DMSO), administered intraperitoneally (i.p.) immediately before bacterial challenge, shortened the survival time of mice infected i.p. with a lethal dose of Salmonella enteritidis NUB 1 (virulent strain) and increased the mortality rate for mice infected i.p. with various bacterial strains showing low virulence for mice, such as S. enteritidis NUB 31 (avirulent strain), Pseudomonas aeruginosa, Klebsiella pneumoniae and Streptococcus pyogenes. It was revealed that DMSO enhanced multiplication of S. enteritidis NUB 1 or facilitated the survival of S. enteritidis NUB 31 in the bodies of mice. At 30 hr after infection with S. enteritidis NUB 1 or NUB 31, most of the viable bacteria contained in the peritoneal fluid were recovered from cells in mice treated with DMSO as well as in untreated mice. There was no evidence that DMSO exerted any direct effect on microorganisms to stimulate in vitro growth or to enhance their virulence for mice. Ingestion by phagocytes of S. enteritidis NUB 1 injected i.p. or intravenously was not inhibited by DMSO. Although DMSO caused a transient decrease in the number of peritoneal macrophages, the enhancing effect of DMSO on infection did not appear to be solely due to a decrease of peritoneal macrophages. The effect of DMSO was highest when it was administered within one hour before or simultaneously with the bacterial challenge. The acceleration of deaths of mice infected with S. enteritidis NUB 1 by DMSO was also observed when the two agents were given orally.
Studies of Pseudomonas aeruginosa Infections Among Hospitalized Patients
Authors: Shigeta Shiro , Ishida Nakao
Published: November 2013

A rapid identification method of glucose nonfermentative gram‐negative rods was established and 320 strains isolated were divided into five groups according to their characteristics in pigmentation, acid from glucose, cytochrome oxidase activity and motility. Further characterization of the strains in each group resulted in the identification that the strains in group I were Pseudomonas aeruginosa, strains in group II, P. aeruginosa and Pseudomonas putida. Achromogenic strains of P. aeruginosa were classified into group III, Pseudomonas maltophilia, Pseudomonas alcaligenes and Alcaligenes faecalis into group IV and Acinetobacter calcoaceticus (Acinetobacter anitratus and Achromobacter lwoffii) in group V. When fluorescent pigment production was taken as a standard, 259 out of 263 chromogenic strains were identified as P. aeruginosa and the remaining four were P. putida. Whereas forty‐five achromogenic strains included twenty‐four A. calcoaceticus, eight P. aeruginosa, six A. faecalis, five P. maltophilia and two P. alcaligenes. From May 1970 to June 1971, 368 strains of glucose nonfermentative rods were isolated from clinical specimens sent to the Central Laboratories of Tohoku University Hospital and three fourth (286/368) of the isolates were P. aeruginosa
A Further Numerical Taxonomic Study of the Rhodochrous Group
Authors: Tsukamura Michio
Published: November 2013

A computer taxonomy was carried out as an aid to clarify the relationships between the genera Mycobacterium, and Nocardia as well as the ‘Rhodochrous’ group. The last group contained slightly acid‐fast organisms isolated from sputa and soil and also included strains received as “Mycobacterium rhodochrous.” The group had been proposed as the genus Gordona by the present author. The rhodochrous group formed a cluster at the 87% similarity level and was clearly separated from all strains of the genera Mycobacterium and Nocardia. The cluster of the rhodochrous group was divided into seven subclusters at the 90% similarity level, and six of the seven corresponded to six species of the genus Gordona: G. bronchialis; G. rubropertincta (synonym G. rubra); G. terrae; G. aurantiaca; G. rhodochroa; G. rosea. If one accepts the concept that a phenetically well‐defined cluster is a species, each of the subclusters of the rhodochrous group should be regarded as a species and the entire group as a genus. Discrepancies between the results of numerical classification and the results of lipid analyses remain as a problem to be solved in the future.
The Effects of Amino Acids and Metals on the Infectivity of Poliovirus Ribonucleic Acid
Authors: Totsuka Atsuko , Ohtaki Kenya
Published: November 2013

Most amino acids, except cysteine, protected the infectivity of phenol‐extracted poliovirus ribonucleic acid against inactivation during incubation at 37 C. Of these amino acids, histidine was most effective, whereas cysteine enhanced inactivation. Besides cysteine, three sulfhydryl compounds were also examined. Thioglycolic acid exhibited a similar effect as cysteine, but glutathione and mercaptoethanol were protective or enhancing in the inactivation of poliovirus RNA depending on the concentration of the drug. The effect of metal ions was diverse. Cu++ and Hg++ reduced the infectivity extensively, whereas Fe3+ and Al3+ were protective. Some of other metal ions were more or less inactivating, while the other metal ions examined were not. A protector and an inactivator of infectious RNA antagonized each other.
Localization of Acid and Alkaline Phosphatases in Staphylococcus aureus
Authors: Okabayashi Kinji , Futai Masamitsu , Mizuno Den'ichi
Published: November 2013

The localization of acid and alkaline phosphatases in Staphylococcus aureus was studied by fractionation of cells after treatment with the L‐11 enzyme and by electron microscopic histochemistry. The two enzyme activities were located in distinctly different positions at the surface of the cells. Acid phosphatase appeared to be localized around the cell membrane of the bacteria, because the enzyme was recovered exclusively in the membrane fraction and because deposition of lead phosphate was detected by electron microscopic histochemistry on the inner surface of the cell membrane of intact bacteria and spheroplasts. The highest specific activity of alkaline phosphatase was also associated with the membrane fraction. However, on electron microscopic histochemistry of intact cells, the deposition of lead phosphate was only seen on the outer surface of the cell wall.
Flagellar Morphology of Vibrio parahaemolyticus (Fujino et al) Sakazaki, Iwanami and Fukumi 19631
Authors: Yabuuchi Eiko , Miwatani Toshio , Takeda Yoshifumi , Arita Michiko
Published: November 2013

The flagellar morphology of 88 Vibrio parahaemolyticus strains, including a strain descended from Fujino's original strain EB101 (= ATCC17802 = KM1339) was studied. EB101 and 83 other strains (95%) showed mixed polar and peritrichous type of flagellation when grown on modified MOF (MMOF) agar after 16‐hr incubation at 20 C. Cultures containing numerous peritrichous cells showed wiggly movements in moist preparations and rapidly spreading growth in semisolid agar plates. Peritrichous flagella were easily removed mechanically from the soma. The mean wavelengths of polar and peritrichous flagella were 2.53 μm (normal type) and 1.72 μm (atypical curly type) respectively. Peritrichous cells on solid media appeared after incubation for 2.5 hr at 37 C and 7 hr at 20 C. Overnight incubation at 37 C and acidity of the medium due to fermentation of carbohydrate markedly ruined peritrichous flagella. Electron micrograph of cells grown on MMOF agar revealed a sheathed polar flagellum and unsheathed peritrichous flagella. A hook structure was demonstrated at the proximal end of the latter. Polar monotrichous cultures in MMOF broth sometimes contained some cells having several or many peritrichous flagella of atypical curly type. Seven strains of Vibrio cholerae were exclusively polar monotrichous on solid and in liquid media. The flagellation of V. parahaemolyticus is concluded as being a mixed polar‐peritrichous type. This fact would indicate that V. parahaemolyticus should be excluded from the genus Vibrio, since the genus Vibrio was defined as polar monotrichous.
Quantitative and Qualitative Analyses of the Antibody Response of Adult Mice Tolerized with Deaggregated Bovine γ‐Globulin
Authors: Hosono Masamichi
Published: November 2013

Mice were made tolerant to bovine γ‐globulin (BGG) by an intravenous injection of deaggregated soluble BGG (sBGG). Two weeks later, they were injected with immunogenic forms of BGG either into foot pads or intravenously. Time course of the number of antibody‐forming cells and the relative avidity of plaque‐forming antibodies in tolerized mice were compared with those of sublethally X‐irradiated mice and of normal immunized mice. The number of plaque‐forming cells (PFC) per popliteal lymph node of tolerant or X‐irradiated mice was shown to be smaller than in the controls. The growth rate of PFC in the nodes in the early logarithmic phase was almost the same between tolerized mice and normal mice, but X‐irradiation resulted in a reduction of the growth rate. Although X‐irradiation did not influence the average avidity of plaque‐forming antibodies, the induction of tolerance by sBGG slightly reduced the avidity in popliteal lymph nodes and conspicuously in spleens about three weeks after the challenge immunization, the relative avidity in the latter being as low as one tenth of the normal immunized mice. In popliteal lymph nodes of tolerized mice at day 9 after immunization, a reduction in the relative avidity of plaque‐forming antibodies was not observed, though the number of PFC was as small as one fifth of the controls. However, X‐irradiation of the hind limbs of tolerized mice prior to immunization caused a reduction in the avidity of plaque‐forming antibodies in the nodes. These results indicated that the precursors of high avidity antibody producers were liable to tolerance induction, and that cells in the recirculating pool were inactivated by the tolerogen more easily than those in such remote sites as the popliteal lymph nodes.
Enhancing Effect of Interferon Pretreatment on Interferon Production
Authors: Ito Fumiaki , Kobayashi Sigeyasu
Published: November 2013

Pretreatment of mouse L cells with interferon (IF) preparations enhanced IF production in response to stimulation with polyinosinic‐polycytidylic acid (poly I·poly C). The enhancement of IF production by the pretreatment was maximal with 5 to 10 units/ml of IF. Pretreatment with approximately 1 unit/ml of IF caused no significant enhancement. The enhancement was observed with all IF preparations tested, crude or partially purified, prepared by induction with either poly I·poly C, Escherichia coli endotoxin or Newcastle disease virus. IFs harvested at different times after induction with high and low concentrations of poly I·poly C were equally effective. The enhancing (“priming”) activity exhibited “species specificity.” However, pretreatment with a high dose of human or rabbit IF, which showed an activity of about 10 units/ml even in the heterologous L cells, enhanced mouse IF production. Studies on physicochemical properties of the active factor indicated that the “priming” activity may be ascribable to IF molecules and not to any other factor in the IF preparations.
In Vitro Synthesis of Reovirus Genomic Segments
Authors: Watanabe Yasushi , Sakuma Sadatoshi , Shames Rose
Published: November 2013

Using a method for labeling RNA with heavy nucleoside 5′‐triphosphates, it was observed that the large particulate fraction isolated from reovirus‐infected cells mediated iv vitro the synthesis of the three size classes of reovirus double‐stranded RNA segments by elongating and terminating minus strand RNA whose synthesis had been initiated in the infected cells. The results are consistent with the hypothesis that each segment of reovirus plus strand template has an individual initiation site for the synthesis of minus strand RNA.
Effects of Chemotherapeutics on Bacterial Ecology in the Water of Ponds and the Intestinal Tracts of Cultured Fish, Ayu (Plecoglossus altivelis)
Authors: Aoki Takashi
Published: November 2013

Drug‐resistant gram‐negative bacilli conferred with R factors were isolated with high frequencies from the intestinal tracts of ayu (Plecoglossus altivelis) cultured in ponds, in which chemotherapeutics had often been used, and with relatively low frequencies from ayu which received no administration of chemotherapeutics. Drug‐resistant bacteria were also isolated at low frequencies from the intestinal tracts of wild ayu in rivers, as well as from the water of ayu‐culturing ponds and some of them carried R factors. The drug‐resistant bacteria carrying R factors were Aeromonas liquefaciens, Citrobacter, Enterobacter cloacae, Escherichia coli, Hafnia and unidentified strains. All the R factors were classified as the Fi–(F) type, except the two R factors detected in an E. coli strain and in an unidentified strain.
Virucidal Effect of Sodium p‐Chloromercuribenzoate on Influenza Viruses Attributable to Inhibition of Virus Particle‐Associated RNA‐Dependent RNA Polymerase
Authors: Shinkai Kenkichi
Published: November 2013

Sodium p‐chloromercuribenzoate (PCMB) caused a noticeable reduction of infectivity of prototype strains of type A and Lee strain of type B influenza viruses at concentrations of 100 and 200 μg/ml, respectively, after an incubation at 37 C for 60 min. The virucidal effect on A/AA/2/60 (H2N2) strain was dependent on the concentration of the drug and temperature as well as on the time of incubation. The reagent exerted this effect at a concentration which induced little change in the hemagglutinating and neuraminidase activities of the virus. PCMB inhibited by 50% the virus particle‐associated RNA polymerase activity of all prototype strains of type A influenza virus at about 2 μg/ml and that of Lee strain of type B influenza virus at 8.5 μg/ml. Other sulfhydryl reagent such as phenylmercuric nitrate also exhibited virucidal effect on A/AA/2/60 virus which paralleled their inhibition of the virus particle‐associated RNA polymerase activity. From these results it was considered likely that the virucidal action of PCMB on influenza viruses was attributable to inhibition of the virus particle‐associated RNA polymerase activity.
Complement Requirement of Neutralizing Antibodies in Different Classes of Immunoglobulin Appearing in Rabbits and Guinea Pigs after Primary and Booster Immunizations with Herpes Simplex Virus
Authors: Shinkai Kenkichi , Yoshino Kamesaburo
Published: November 2013

Rabbits and guinea pigs were immunized with herpes simplex virus and bled periodically. The sera were fractionated into slow IgG, fast IgG and IgM by DEAE‐cellulose column chromatography, and complement‐requiring (CRN) and nonrequiring neutralizing (N) antibody activities were estimated. In early sera of rabbits, the two IgG and IgM fractions possessed about equal CRN activities, although some animals showed a slightly lower activity in fast IgG. In guinea pigs, the early CRN activity resided mainly in slow IgG (7 S γ2). The early IgG antibody of guinea pigs differed from that of rabbits in that it resembled IgM in resistances to heating at 70 C and to 2‐mercaptoethanol. The level of CRN IgM antibody in rabbits declined following a peak reached in 2 to 3 weeks, whereas such a decline was never observed in guinea pigs. N IgG antibody was developed a few weeks after the first immunization in rabbits and much retarded in guinea pigs. In both species, booster immunization quickly evoked N antibody in the two IgG fractions and also CRN IgM antibody, but in the case of rabbits the IgM antibody disappeared soon. It is concluded that IgG plays an important role in humoral immunity from the initial stage of the immunization course.
Studies on Gonococcal Infection
Authors: Ota Fusao , Morita Junji , Yoshida Nagayuki , Ashton Fraser , Diena Benito
Published: November 2013

Macrophages from mice were infected with Neisseria gonorrhoeae type 1 cells, and their ultrastructure was studied by electron microscopy. The macrophages showed various stages of engulfment and digestion of gonococci 2 hr after infection. Infected macrophages seemed to develop pseudopodia for phagocytosis, and could engulf more than 30 gonococcal cells. Some engulfed bacteria appeared morphologically intact, while others appeared lysed and some structures resembling the L form of N. gonorrhoeae were also seen. These observations suggest that gonococcal cells may be able to survive intracellularly with normal or altered forms of morphology, and that macrophages containing these bacteria may disseminate gonococcal infection in man.
An Amber Suppressor of Escherichia coli Strain KO1
Authors: Aoi Takeshi , Watanabe Itaru
Published: November 2013

The suppression characteristics of Escherichia coli strain KO1 have been investigated. The growth patterns of nonsense mutants of RNA (GA and f2) and DNA (λ and T4) phages suggested that KO1 carried an amber, but not ochre or opal suppressors. The comparison of KO1 with previously identified amber suppressors indicated that KO1 differed from su1, su3 and su6 in its suppression pattern. KO1 and su2 shared some properties in common, for instance, their ability to suppress GA amber mutants with one exception (amN20) and the restriction of suppression capacity by the strr mutation. However, the suppression efficiency of KO1 (48%) was about three times that of su2 (18%). A possibility that KO1 contained a new amber suppressor is discussed.
Adjuvant Activity of Mycobacterial Fractions
Authors: Taniyama Tadayoshi , Azuma Ichiro , Yamamura Yuichi
Published: November 2013

A quantitative assay and characterization of oil‐attached cell wall of Mycobacterium bovis BCG (BCG‐CWS) which stimulates cell‐mediated immunity of spleen cells to alloantigens in mice were carried out by an in vitro cell‐mediated cytotoxicity test using 51Cr‐labeled target cells. C57BL/6J mice (H‐2b) were immunized intraperitoneally with mastocytoma cells (H‐2d) with or without oil‐attached BCG‐CWS. The cytotoxicity, comparable to that of spleen cells from mice immunized with mastocytoma cells (3 × 107), could be induced in spleens of mice immunized with a mixture of mastocytoma cells (104) and oil‐attached BCG‐CWS. The enhancing effect persisted for 55 days or more after the alloantigenic immunization. Oil‐attached BCG‐CWS enhanced cell‐mediated cytotoxicity of T cells in the spleen and the mesenteric lymph node, but not in the thymus. The cytotoxicity showed specificity toward the alloantigen used for immunization. In addition to BCG‐CWS, the cell walls of Nocardia rubra and Corynebacterium diphtheriae PW8 and the peptidoglycolipids of Mycobacterium tuberculosis Aoyama B were found to be potent stimulants of cell‐mediated cytotoxicity in mice. Oil‐attached BCG‐CWS did not enhance humoral response to mastocytoma cells but enhanced cell‐mediated cytotoxicity when viable mastocytoma cells were used as antigen. The above result was supported by the fact that anti‐hapten antibody response induced by viable trinitrophenyl (TNP)‐mastocytoma cells (104) plus oil‐attached BCG‐CWS did not increase to the maximum level as was observed in mice immunized with a larger number of mastocytoma cells (3 × 107) alone, while cell‐mediated cytotoxicity induced by the same treatment increased to the maximum level obtained by immunization with mastocytoma cells (3 × 107) alone.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Nakashima Izumi , Kato Nobuo
Published: November 2013

A study was performed to clarify the roles of primary and secondary injections of antigen and adjuvant (capsular polysaccharide of Klebsiella pneumoniae, CPS‐K) in induction of antibody responses and in development of immunological memory in mice to bovine serum albumin (BSA). A primary injecion of BSA alone neither induced significant primary antibody response nor increased immunological memory for a secondary antibody response but, if primary injections of BSA and CPS‐K were performed simultaneously, high antibody responses were induced. Moreover, a prior injection of BSA alone or CPS‐K alone decreased the level of primary antibody response and the degree of increase in memory following the subsequent injection of BSA mixed with CPS‐K. In contrast, a secondary injection of BSA alone into mice once primed with a mixture of BSA and CPS‐K elicited very high secondary type antibody response and increased secondarily the memory for a tertiary antibody response. Injection of CPS‐K simultaneously with or shortly before or after the secondary injection of BSA did not increase the level of the secondary antibody response and the degree of the secondary increase in memory. Augmentation of the secondary antibody response was elicited by simultaneous injection of CPS‐K only when the secondary response was induced inadequately by a suboptimum or supraoptimum dose of antigen.
R Factor‐Mediated Resistance to Aminoglycoside Antibiotics in Pseudomonas aeruginosa
Authors: Sagai Hitoshi , Krcmery V. , Hasuda Katsumi , Iyobe Shizuko , Knothe H. , Mitsuhashi Susumu
Published: November 2013

Conjugal transferability of drug resistance was examined, in eleven Pseudomonas aeruginosa strains which were isolated in Frankfurt. Four R factors were demonstrated from three strains using P. aeruginosa as recipients but they were nontransferable to Escherichia coli K12. Two R factors, i.e., Rms146 and Rms147, mediated resistances to tetracycline (TC), streptomycin (SM), sulfanilamide (SA), kanamycin (KM), lividomycin (LV), gentamicin C complex (GM) and 3′,4′‐dideoxykanamycin B (DKB). They mediated the formation of aminoglycoside‐inactivating enzymes, i.e., SM phosphotransferase, SM adenylyltransferase, KM and LV phosphotransferase 1, and GM and DKB 6′‐N‐acetyltransferase. TC resistance conferred by these R factors was due to impermeability of the drug. P. aeruginosa Ps 142 carried two kinds of R factor in one cell, Rms148 (SM) and Rms149 (SM·SA·GM·CPC) (CPC, carbenicillin). Rms148 (SM) was transferable at a high frequency of 10–1 and mediated the formation of SM phosphotransferase. Rms149 mediated the formation of drug‐inactivating enzymes, i.e., GM 3‐N‐acetyltransferase and β‐lactamase, but did not inactivate SM. SM resistance was probably due to impermeability of the drug.
Eliminatory Action of Glycine on Drug Resistance of Escherichia coli K12 Harboring an R Factor
Authors: Tomoeda Munemitsu , Inuzuka Manabu , Hayashi Masako
Published: November 2013

Glycine, known to inhibit the synthesis of a peptidoglycan component of the bacterial cell wall, was effective in eliminating drug resistance of Escherichia coli K12 JE2100 strain harboring the R100–1 factor, although in lower frequencies than that of sodium dodecyl sulfate (SDS). The action of glycine was found to be less effective on the same R factor in JE177 strain, and not effective on the F factor in W6. Infection of R factors from R+ cells to R– cells was found to take place in the glycine broth as efficiently as in broth without glycine. This might result in lowering the apparent efficiency of the action of glycine on those plasmids. The segregation patterns of drug‐susceptible clones obtained by the glycine treatment were different from those obtained after the SDS treatment. These results coupled with other evidences suggest that the mode of action of glycine on R+ cells may be different from those of other curing agents and may involve mechanisms other than selection of R– or drug‐susceptible segregants that are present in R+ culture.
Epidemiological Survey of Streptococcus mutans among Japanese Children
Authors: Hamada Shigeyuki , Masuda Norio , Ooshima Takashi , Sobue Shizuo , Kotani Shozo
Published: November 2013

An epidemiological investigation was carried out to identify and determine the serotypes of Streptococcus mutans from carious lesions of young Japanese children. For this purpose, a direct fluorescent antibody technique was mainly used. Fluorescein isothiocyanate‐conjugated antibodies were prepared for the five known serotypes of S. mutans. Cross reactions and nonspecific reactions were eliminated by adsorption, counterstaining, or DEAE‐cellulose column chromatography. Agar‐gel immunodiffusion was used to distinguish between serotypes a and d. The epidemiological survey suggested that serotype c strains were most prevalent in dental plaques of Japanese children. The d and e serotypes were rare and serotypes a and b were not detected. It was also noted that more than one serotype of S. mutans could be found in the same locus of a carious lesion and that there might be no relationship between the degree of caries and the causative serotype(s) of S. mutans.
In Vitro Studies on the Mechanism of Acquired Resistance to Tuberculous Infection
Authors: Muraoka Shizuko , Takeya Kenji , Nomoto Kikuo
Published: November 2013

The relationship between lymphocytes and macrophages in cellular immunity against tuberculous infection was studied by means of an in vitro cell culture system without addition of streptomycin. The peritoneal macrophages were obtained from normal mice or mice immunized with heat‐killed tubercle bacilli in paraffin oil, boosted with live BCG and infected with H37Rv cells in vitro. The infected monolayers of macrophages were cultivated for 48 hr with immune lymphoid cells obtained from immunized mice. The intracellular growth of H37Rv cells 3,5 and 7 days after infection was examined by counting tubercle bacilli within infected macrophages under a microscope. 1) The increase of bacilli within macrophages derived from immunized mice was slightly smaller than that in normal macrophages. 2) The addition of immune lymph node cells to the macrophage monolayers resulted in a marked decrease in the number of bacilli within both normal and “immune” macrophages. Conversely, normal lymph node cells exhibited an enhancing effect on the intracellular bacillary growth. 3) Immune lymph node cells showed a higher capacity to cause macrophages to suppress intracellular growth of bacilli than that of splenic lymphoid cells or thymocytes after addition to macrophage monolayers. 4) The treatment of lymphoid cells with inhibitors of protein synthesis, cycloheximide or streptovitacin A, resulted in a remarkable reduction of the ability of sensitized lymphocytes to cause macrophages to suppress multiplication of intracellular bacilli.
Application of the Macrophage Migration Inhibition Test to Screen Patients with Early Cancer and Obtain Prognostic Determinations of Cancer Treatment
Authors: Akiyama Takehisa , Yamaura Noboru
Published: November 2013

A modified direct method of the macrophage migration inhibition test (MMIT) was attempted on a large number of patients with malignant or benign tumors. Results of the MMIT in almost all patients with benign tumors were negative except for those with hydatidiform moles, dermoid cysts and viral benign tumors such as verruca plana which were positive. The number of cases determined as false positives were exceptionally few. Conversely, about the half of the patients with malignant tumors were positive. The majority of negative cancer patients were confirmed pathologically to be advanced cases and, therefore, were postulated to have been immunologically unresponsive. The remaining false negative patients were diagnosed to be very early cases with their malignant foci too small to be effective antigenic stimuli. The MMIT was also performed postoperatively on some of the patients using autologous antigens, which had been preserved by freezing, for examination of changes in the per cent migration index. The results led the authors to conclude that postoperative repetitions of the test permitted them to tell that cancer cells had been completely eradicated or that a relapse might occur in the near future. Examinations of cross reactivity between tumor antigens revealed that such reactivity exists between cancer antigens and antigens originating in hydatidiform moles and that there is also a very strong cross reactivity between allogeneic cancer antigens regardless of differences in the organs of origin. This fact suggests that the present test is effective for the screening of preoperative patients with early cancer.
Microorganisms Responsible for Controlling the Populations of Escherichia coli and Enterococcus and the Consistency of Cecal Contents in the Chicken
Authors: Morishita Yoshiyuki , Mitsuoka Tomotari
Published: November 2013

A study was made to clarify what kinds of intestinal organisms might be responsible for controlling the populations of Escherichia coli and Streptococcus faecalis var. liquefaciens in the cecum and the consistency of the cecal contents of chickens. Germ‐free chickens were inoculated orally with various mixtures of bacterial cultures alone or in combination, different dilutions of the cecal contents of chickens, different dilutions of the cecal contents treated by heating or with chloroform, the supernatant of diluted cecal contents, and dilutions of human feces. Factors controlling the E. coli population, enterococcal population, and consistency of the cecal contents were shown to be independent of one another. The ecosystem controlling the E. coli or enterococcal population was more complex than that controlling the consistency of the cecal contents. The former was composed of anaerobic and facultatively anaerobic bacteria isolated and heat‐ or chloroform‐resistant organisms, and the latter of heat‐ or chloroform‐resistant organisms alone, which were inferred not to be prevailing in the cecal contents of chickens. Discussion is made on ecological systems controlling the intestinal flora.
Purification and Properties of Glucose‐6‐Phosphate Dehydrogenase from Bacillus subtilis Spores
Authors: Tanahashi Takako , Tochikubo Kunio , Hachisuka Yoetsu
Published: November 2013

Glucose‐6‐phosphate dehydrogenase [d‐glucose‐6‐phosphate: NADP oxidoreductase, EC. 1. 1. 1. 49] obtained from spores of Bacillus subtilis PCI 219 strain was partially purified by filtration on Sephadex G‐200, ammonium sulfate fractionation and chromatography on DEAE‐Sephadex A‐25 (about 54‐fold). The optimum pH for stability of this enzyme was about 6.3 and the optimum pH for the reaction about 8.3. The apparent Km values of the enzyme were 5.7 × 10–4 M for glucose‐6‐phosphate and 2.4 × 10–4 M for nicotinamide adenine dinucleotide phosphate (NADP). The isoelectric point was about pH 3.9. The enzyme activity was unaffected by the addition of Mg++ or Ca++. The inactive glucoses‐6‐phosphate dehydrogenase obtained from the spores heated at 85 C for 30 min was not reactivated by the addition of ethylenediaminetetraacetic acid, dipicolinic acid or some salts unlike inactive glucose dehydrogenase.
Persistent High Numbers of Clostridium perfringens in the Intestines of Japanese Aged Adults
Authors: Yamagishi Takayoshi , Serikawa Toshihiko , Morita Ryoko , Nakamura Shinichi , Nishida Shoki
Published: November 2013

TSN agar was applicable for enumeration of Clostridium perfringens in fecal samples of adults but not in those of infants. It was demonstrated using TSN agar that some healthy aged adults had persistently carried C. perfringens at levels ranging from 107 to 109, while some others ranged from 103 to 106 per ml volume of fecal sample although all of these adults had the same diets. In the test for agglutinability of isolates of C. perfringens collected from two elderly adults, a younger adult and a baby, it was demonstrated that most of the isolates obtained from an aged adult of high levels for 19 months belonged the same serotype, while rapid alteration of serotypes could be observed in three other persons with high or low levels. In spite of as many as 109 C. perfringens per ml of feces, no trace of α‐toxin could be detected in the fecal samples. In in vitro tests, fecal suspension suppressed the production of α‐toxin although it allowed the organism to grow sufficiently.
Effect of Capsular Polysaccharide of Klebsiella pneumoniae on Host Resistance to Bacterial Infections
Authors: Kato Nobuo , Kato Osamu , Nakashima Izumi
Published: November 2013

In normal mice, the total count of peritoneal leukocytes was markedly decreased after intraperitoneal (i.p.) injection of the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) depending on the dosage injected. This decrease was mainly due to the depletion of macrophages, and a decrease in the number of lymphocytes occurred to a lesser extent. CPS‐K in relatively smaller doses mobilized polymorphonuclear neutrophilic leukocytes (PMN) into the peritoneal fluid but it decreased them transiently in larger doses. In mice infected i.p. with a virulent strain of Salmonella enteritidis, there was an abundant emigration of PMN into the peritoneal fluid. When 200 μg of CPS‐K was injected i.p. immediately before bacterial challenge, emigration of PMN was markedly delayed for 48 hr after infection. Associated with this suppressed emigration of PMN, the numbers of macrophages and lymphocytes in the peritoneal fluid were significantly less in mice treated with CPS‐K than those in untreated control mice for 48 hr after infection. The numbers of both cell‐associated and extracellular bacteria in the peritoneal fluid were markedly greater in mice treated with CPS‐K than those in untreated control mice. In both in vivo and in vitro experiments, ingestion of bacteria by macrophages and PMN was not blocked by CPS‐K or neutral CPS‐K, the active substance responsible for the infection‐promoting effect of CPS‐K. It appeared that CPS‐K somehow impaired the intraphagocytic bactericidal activity.
Multiple Mating Types of Mycobacterium smegmatis
Authors: Mizuguchi Yasuo , Suga Kiyoko , Tokunaga Tohru
Published: November 2013

Multiple mating systems were found in the strains of Mycobacterium smegmatis. Nineteen wild type strains were classified into five different groups according to their mating behavior. All crosses between substrains derived from the same wild type strain or between substrains belonging to the same group were infertile, while eight different intergroup crosses were fertile. It was suggested that in most of the fertile crosses, chromosome transfer was unidirectional. Genes controlling matings were resistant to curing by acridine dye, sodium dodecyl sulfate and higher temperature suggesting that they were chromosomal genes. However, the location of these genes could not be demonstrated because none of the chromosomal markers tested was found to be linked to these mating genes.
Cytoplasmic Budding of a Nuclear Polyhedrosis Virus and Comparative Ultrastructural Studies of Envelopes
Authors: Kawamoto Fumihiko , Suto Chiharu , Kumada Nobuo , Kobayashi Masahiko
Published: November 2013

The processes of cytoplasmic budding in Euproctis subflava nuclear polyhedrosis virus (NPV) were investigated, and comparisons were made among three types of envelopes which were acquired by, 1) de novo morphogenesis in the nuclei, 2) nuclear budding, and 3) cytoplasmic budding. The direction of nucleocapsids in the envelope was the same in these three modes of envelopment; the envelopment seemed to occur from a nipple end which was at one extremity of the nucleocapsid. After the envelopment, electron‐dense materials were seen between the envelope and nucleocapsid, though their contents and morphological features were different among the three types of envelopes. However, these materials seemed to function similarly as a mediator between the envelope and nucleocapsid as have been observed in many vertebrate viruses which acquire envelopes.

A marked difference among the three types of envelope was the characteristic cap‐shaped structures with spikes which were seen only on the surface of envelope derived from the plasma membrane. After cytoplasmic budding, nucleocapsids enveloped by this way were located on the basement membrane or liberated in the hemocoel, and then they appeared to enter neighboring healthy cells via viropexis with the spike end at the head. At the sites where these spikes came into contact with healthy cells, coated vesicle‐like structures were observed inside the plasma membrane. Occasionaly, incomplete particles which lacked nucleocapsids were also budded through the plasma membrane and released into extracellular space.
Fate of Tobacco Mosaic Virus after Entering the Host Cell
Authors: Hayashi Takaharu
Published: November 2013

Diminutive viral RNAs recovered from tobacco leaves inoculated with 32P‐TMV were investigated. At 3.5 hr after inoculation, most of the viral RNA without coat protein revealed two peaks after sucrose density gradient analysis of SDS‐extract from 12,000 × g leaf pellet. The first peak appeared between bacterial ribosomal RNA of 16 S and 5 S and the second peak was around 5 S. These two peaks were digestible with RNase and they appeared as early as 5 min after inoculation. These diminutive RNAs seemed to be derived from partially uncoated parental virus by abscission of their naked RNA tails. The active formation of these diminutive RNAs and their early appearance after inoculation seemed to indicate that most of the inoculated TMV received incomplete uncoating.