bluuebunny/crossref_metadata_2025 huggingface dataset:
Factors Influencing Competence of Escherichia coli for Lambda‐Phage Deoxyribonucleic Acid Infection
Authors: Higa Akiko , Mandel Morton
Published: November 2013

Under certain conditions, Escherichia coli K12 cells lysogenic for λ can be made sensitive to free phage DNA without the addition of external helper phage. This competence depends on the phages released in the growth medium by spontaneous lysis. The kinetics of the decay of this competence are similar to the kinetics of cyclization of injected phage DNA.
Studies on γM and γG Antibody Response of Mice to Bovine Serum Albumin
Authors: Katsura Yoshimoto
Published: November 2013

Tolerogenic effect of soluble deaggregated bovine serum albumin (sBSA) on the γM and γG antibody responses was studied. Normal and tolerogen‐treated animals were immunized with the intravenous injection of alum‐precipitated BSA (net 0.1 mg) and a bacterial endotoxin (0.01 mg), since this immunizing procedure was the most adequate to elicit the strong responses of both types of antibodies. Both γM and γG responses were decreased evenly by a single injection of small dose (0.01 or 0.1 mg) of sBSA. Larger doses (1 mg or 5 mg) caused the marked suppression of γM response, but the γG response was reduced only slightly. The tolerant state was facilitated by weekly injections of 0.1 mg of sBSA, both γM and γG responses being abrogated profoundly by more than three injections. The induction of such a complete tolerance was not ascribed solely to either the total dose or the period of time between the first tolerogen injection and the challenge immunization, but to the retention of antigen more than a certain threshold amount in the body for some duration.
Further Simplification of the Serum Neutralization Test with Herpes Simplex Virus
Authors: Hondo Ryo , Yoshino Kamesaburo
Published: November 2013

A new, simple technique for the serum neutralization test with herpes simplex virus was devised. Thin discs made of solidified overlay medium containing about 2×106 PFU/ml of virus (VACS) were covered with 0.01 ml of graded serum dilutions or control saline and placed on chick embryo cells with the serum‐covered face down. A 90‐mm monolayer dish received 8 such VACSs and then fixing overlay. The highest dilution of the serum which reduced confluent plaques to no or a few scattered plaques was taken as the endpoint. Incubation of the serum‐covered VACSs at 37 C for 1 hr prior to inoculation could be omitted, since this procedure did not markedly increase the endpoint. A 2‐ to 4‐fold fluctuation of the virus concentration had little influence on the endpoint. The standard deviation of the endpoint obtained was within ±2‐fold. The sensitivity of the test was about one‐third that of the 50% plaque reduction method. Early sera containing predominantly complement‐requiring neutralizing antibody exhibited considerable titers when tested by the new method even in the absence of complement, the reason for which remains unknown.
Enhancement of Endotoxin‐Induced Virus‐Inhibiting Factor or Interferon Production by Pretreatment of the Rabbit with Newcastle Disease Virus
Authors: Nagano Yasuiti , Maehara Nobutoshi
Published: November 2013

Rabbits pretreated by intravenous injection with Newcastle disease virus (NDV) produced high‐titered virus‐inhibiting factor (IF) or interferon in the serum upon intravenous injection of Escherichia coli endotoxin 24 hr later as compared with the IF production induced by endotoxin in rabbits without the NDV pretreatment. IF assays of different organs revealed that the NDV pretreatment rendered the spleen rather hyporeactive but other organs such as the liver, lungs and kidneys hyperreactive to the subsequent IF induction by endotoxin. The enhanced IF titer observed in the serum seems to be the summation of IF released from various organs rendered hyporeactive or hyperreactive by the NDV pretreatment. It is postulated that the late‐appearing, heat‐ and acid‐stable IF and the early‐appearing, heat‐ and acid‐labile IF are produced by different processes in different types of cells. The production of early IF consists of two steps, formation of “preinterferon” and conversion of preinterferon to early IF. Preinterferon formation is induced by virus but not by endotoxin, while the conversion of preinterferon to early IF is effected by both endotoxin and virus. The formation of late IF is induced only by virus, taking a one‐step process. This hypothesis seems to explain the findings in the present study.
Drug Resistance to Aminoglycosidic Antibiotics in Pseudomonas aeruginosa and Its Lability
Authors: Kobayashi Fujio , Yamaguchi Masahito , Mitsuhashi Susumu
Published: November 2013

Antibacterial activity of aminoglycosides toward Pseudomonas aeruginosa was investigated by using 221 clinical isolates. One hundred and seventy‐five strains were shown to be highly resistant against one or more of the antibiotics tested. Aminoglycoside resistance was found to be lost in some strains of pseudomonads, suggesting that the determinants would be located extrachromosomally. Loss of resistance in some strains was paralleled with the loss of an ability to inactivate the drug.
Antibacterial Activity of Chloramphenicol‐Related Compounds toward a Chloramphenicol‐Resistant Strains of Staphylococcus aureus
Authors: Kono Megumi , O'Hara Koji , Nagawa Masatoshi , Mitsuhashi Susumu
Published: November 2013

Antibacterial activities of 146 chloramphenicol (CM) derivatives and analogues were investigated in a CM‐resistant strain of Staphylococcus aureus. Seventeen compounds tested showed the same or much higher antibacterial activity when compared with CM. These compounds could be classified into two groups: The first group included ten compounds containing a benzoyl moiety in their structure and the remaining seven compounds did not possess that moiety. Compounds, which showed a fourfold higher antibacterial activity than CM, were six CM derivatives and two CM analogues. They were all included in the first group. Present and previous studies indicated that the most active CM‐related compound was DL‐α‐dichloroacet‐amido‐α‐methylene‐p‐nitropropiophenone, termed No. 61 (previously called CM 17). The mechanism of antibacterial activity of this compound was due to the fact that this was not inactivated by the CM acetyltransferase produced by the CM‐resistant strain.
Application of Cell Culture in Studying Antibacterial Activity of Rifampicin toShigellaand EnteropathogenicEscherichia coli
Authors: Osada Yasuaki , Nakajo Masayuki , Une Tsutomu , Ogawa Hidemasa , Oshima Yasuo
Published: November 2013

Antibacterial activity of rifampicin (RFP) against intracellularShigellaand enteropathogenicEscherichia coliwas tested in a cell culture system and compared with that of known active controls such as chloramphenicol (CP), kanamycin (KM), streptomycin (SM) or tetracycline (TC), with reference to minimal inhibitory concentrations (MIC) and to chemotherapeutic effects on intraperitoneal infections in mice. When RFP, CP, or TC was applied to L cells and HeLa cells infected respectively withShigella flexneri2a and enteropathogenicE. coli0143, both cell‐infection and intracellular multiplication of the bacilli were considerably inhibited, while RFP was less active against the extracellular bacilli in cell‐free environments than KM and SM. Comparing the intracellular antibacterial activity between RFP and TC or CP, the former was less active than the latter two. This discrepancy in antibacterial activity between extra‐ and intracellular environments could in part be attributed to the fact that RFP penetrated more easily into cultured cells than KM and SM did.
Detection of R+ Bacteria in Cultured Marine Fish, Yellowtails (Seriola quinqueradiata)
Authors: Aoki Takashi , Egusa Syuzo , Watanabe Tsutomu
Published: November 2013

Many drug‐resistant gram‐negative bacilli were isolated from the intestinal tracts of yellowtails (Seriola quinqueradiata) cultured on farms in various parts of Kochi and Ehime Prefectures, Shikoku Island. They were Vibrio, Pseudomonas, Proteus, Citrobacter and some unidentified Enterobacteriaceae. Of these drug‐resistant strains, Vibrio and Pseudomonas were found to carry R factors in high frequencies. These R factors had four types of resistance markers, SA, SA. CM, SM. CM. ABP. and SA. SM. CM. TC. All R factors were found to belong to the fi– type. In contrast, only one drug‐resistant gram‐negative bacillus was detected in a cultured yellowtail on farms near Numazu City, Shizuoka Prefecture.
Dose Effect of Immune Ribonucleic Acid Fraction on Antibody Formation
Authors: Kitamura Kouichi , Kurashige Satonori , Mitsuhashi Susumu
Published: November 2013

An immune ribonucleic acid (iRNA) preparation was extracted with phenol from the spleens of mice immunized with sheep red blood cells (SRBC). When mice were injected intravenously with iRNA, rosette formers in the spleens of such mice were demonstrated but plaque formers and serum antibody could not be detected. As reported previously, iRNA against SRBC also could induce memory cells capable of responding to a small amount of antigen to produce a high titer of antibody. The necessary interval between the iRNA and antigen stimulation for induction of a secondary response was found to be three weeks or more. Presence of an optimal dose of iRNA for the induction of memory cells was demonstrated, and the role of iRNA in antibody formation and the optimal dose of iRNA for the induction of memory cells were discussed.
The Possible Presence of Ribonucleic Acid‐Dependent Deoxyribonucleic Acid Polymerase in the Immune Response
Authors: Kurashige Satonori , Mitsuhashi Susumu
Published: November 2013

Immune ribonucleic acid (iRNA) preparations were extracted from spleens of rabbits immunized either with Salmonella flagella or f2 phage. The enzyme extract prepared from popliteal lymph node cells of immunized rabbits induced DNA synthesis in a cell‐free system in the presence of iRNA. The enzyme activity was demonstrated only after stimulation with a corresponding antigen and was inhibited by large amounts of rifampicin, rifampicin derivatives or by treatment of the iRNA with ribonuclease, but neither actinomycin D nor mitomycin C affected it. Pretreatment of iRNA with deoxyribonuclease did not affect the enzyme activity. This DNA polymerase could induce DNA synthesis only by using the corresponding iRNA but not by using either normal RNA or iRNA prepared against another antigen. This new DNA presumably represents an intermediate in the antibody formation process specified by this enzyme using iRNA as a template.
Immunochemical Studies on Bacterial Blood Group Substances
Authors: Kochibe Naohisa , Furukawa Ken , Iseki Shoei
Published: November 2013

Two oligosaccharides were isolated from a partial hydrolysis of the lipopolysaccharide from Shigella dysenteriae which had a common antigen with human red blood cells (RBC). A disaccharide sh‐2 had a structure of O‐β‐D‐galactosyl‐(l→2)‐D‐galactose, and was active in the hemagglutination inhibition test between human O RBC and anti‐S. dysenteriae chicken serum. A tetrasaccharide sh‐12 was composed of galactose and rhamnose and was a more effective inhibitor than sh‐2. Both oligosaccharides were assumed to be derived from the determinant structure of a common antigen. Serological specificity of the hemagglutinin in chicken serum was examined using these oligosaccharides and human milk oligosaccharides, and the 1→2 linked galactosyl residue was supposed to be important for determination of the heterophile RBC antigenicity.
Purification and Characterization of Mouse L Cell Interferon
Authors: Kawade Yoshimi
Published: November 2013

Interferon was produced in high yields in mouse L cell cultures infected with Newcastle disease virus, with a specific activity of the order of 106 units per mg protein. It was partially purified by zinc acetate precipitation, carboxymethyl Sephadex chromatography, Sephadex gel filtration and pressure dialysis. On electrophoresis in polyacrylamide gel, it consisted of a fast‐moving sharp component and a slow, broadly distributed component(s). The highest specific activity of the former component so far obtained was 8 × 107 units per mg protein, numerically the highest value ever reported for interferon. It was considered likely, however, that the protein components in the purified samples, revealed by staining of the electrophoresis gel, still represented mostly impurities. Gel filtration experiment indicated some heterogeneity of interferon in molecular weight but the major component was estimated to be 30 000 in molecular weight. Interferon activity could be maintained without added preservatives for prolonged periods, provided that the protein concentration of the sample itself was high. One interferon unit as defined in this paper was found to correspond to 0.3 international unit.
Formation of the Recombinant between Ampicillin Re istance Determinant and Transfer Factor
Authors: Nakajima Takashi , Suzuki Mitsue , Kameda Mitsuo , Harada Kenji , Mitsuhashi Susumu
Published: November 2013

In vitro‐developed resistant mutants were obtained by inoculating a clinical isolate of Aerobacter cloacae on plates containing various concentrations of ampicillin (APC). This resistance was paralleled by an increase in the formation of β‐lactamase. When A. cloacae carrying the nontransmissible APC resistance‐determinant (amp) was infected with a T‐tet factor, a recombinant T‐tet.amp factor was formed; the recombinant being conjugally transmissible and capable of conferring APC resistance on their host. This fact implies the origin of the R factor; the R factor being formed by the recombination of sex factors with nontransmissible drug resistance‐determinants.
Development of a Sensitive Microbioassay Using Amplified Cultivation1
Authors: Nakamura Hiroshi , Samejima Keijiro , Tamura Zenzo
Published: November 2013

A novel ultramicro microbioassay was developed. The present method, referred to as amplified cultivation, is composed of two successive kinds of culture. The first culture is performed until a maximum difference is obtained in the ratio of numbers of viable cells appearing in a basal medium and that contained in a nutrient. The second culture is continued after adding a complete medium to the cultures and stopped just after the growth of microorganisms in the blank reaches a certain detectable limit, e.g., by optical density or acid production. Lactose was analyzed by this amplified cultivation method with an extremely low concentration of viable cells of Bifidobacterium bifidum N4 as the inoculum. The detectable limits of lactose were 0.05 μg and 1 μg by the test tube method and by the pulp disc method respectively, with an increase in sensitivity by a factor of more than 103 compared with the corresponding conventional microbioassays. Moreover, a relationship between the diameters of the growth zones and the logarithm of the amount of lactose was achieved in a range of 50–1000 μg of lactose in the pulp disc method.
Detection of a Specific Surface Antigen(s) in Adenovirus Type 12‐Transformed Cells by Fluorescent Antibody Technique
Authors: Hamada Chuya , Nakajima Setsuko , Uetake Hisao
Published: November 2013

A surface antigen(s) was detected in adenovirus type 12 (Ad12)‐transformed cells by fluorescent antibody technique. The antigen was revealed by antisera of mice and rabbits immunized with the virus, and by those of rabbits immunized with conditioned medium from cell cultures carrying the surface antigen. The antigen was specific for the Ad12‐transformed cells, since it was found only in the Ad12‐transformed cells regardless of the animal species and not in any other cells, including SV40‐ and polyoma virus‐transformed cells. The antigen was distinct from the Ad12‐specific viral and T antigens, since the sera of rabbits immunized with the conditioned medium contained little or no detectable antibodies to the latter two antigens. On the contrary, the surface antigen was not detected in any tumor cells induced by Ad12 by the above antisera, and no detectable antibody to the surface antigen was produced in mice immunized with the tumor cells.
Opsonization of Staphylococcus aureus by Guinea Pig 7S Gamma‐Globulin
Authors: Downey Ronald J.
Published: November 2013

Opsonization of Staphylococcus aureus to phagocytosis by guinea pig peritoneal exudate cells (PEC) was examined with various isolated serum globulins. Significant enhancement of in vitro phagocytosis was afforded by partially purified 7Sγ1‐immunoglobulin. The rate of killing or the rate of ingestion of 3H‐thymidine labeled bacteria by PEC was used as an index of phagocytosis. The results indicate that the immunoglobulin can mediate the adsorption of an allogeneic surface independent of a specific antigenic component.
Factors in the Fermentation‐Inhibition Test for Measurement of Growth‐Inhibiting Antibody to Mycoplasma pneumoniae
Authors: Hasegawa Sumio
Published: November 2013

Factors in the fermentation‐inhibition test for the measurement of growth‐inhibiting antibody in serum to Mycoplasma pneumoniae were studied. The fermentation‐inhibiting antibody titer, as read on the day when the color of the control cup without serum had just changed to yellow, was constant among inoculum dilutions of 10–2 to 10–5 (106 to 103 CFU/ml) of a M. pneumoniae broth culture. The use of 10–3 to 10–5 dilutions (105 to 103 CFU/ml) was adequate for inoculation, inasmuch as one day delay in reading did not result in a significant decrease in the test. Heat inactivation of the serum gave no significant effect on the titer. The test was simple and reliable. The growth‐inhibiting antibody was shown to be detectable in the test, when the growth of M. pneumoniae was suppressed at least to 1/100 of the growth of the control without serum. The growth‐inhibiting antibody titer rose later than the complement‐fixing antibody titer in some cases after M. pneumoniae infection. It is suggested that, when an erythromycin‐sensitive strain of M. pneumoniae is used, the titer transiently rises and does not show a real growth‐inhibiting antibody titer in sera from patients under erythromycin administration.
Phenotypic Mixing of Vesicular Stomatitis Virus with HVJ (Sendai Virus)
Authors: Kimura Yoshinobu
Published: November 2013

Vesicular stomatitis virus (VSV) plaque‐forming virions obtained from mixed infection of HeLa cells with VSV and HVJ (Sendai virus) were shown to contain two types of phenotypically mixed virions: virions doubly neutralizable with both anti‐VSV and anti‐HVJ antisera (up to 15%), and virions which could be neutralized only with anti‐HVJ serum and not with anti‐VSV serum (up to 0.3%). No evidence for genetic recombination was obtained between these two viruses. Phenotypically mixed virions could adsorb to and elute from chiken erythrocytes, and bullet‐shaped virions, typical of VSV, were found to be adsorbed to chicken erythrocytes in electron microscopy. Neuraminidase activity was detected in the virions specifically sedimented with anti‐VSV serum. From these findings, phenotypically mixed virions were suggested to contain hemagglutinin and neuraminidase of HVJ in the envelope. In sucrose gradient centrifugation, phenotypically mixed virions enclosed within HVJ antigens sedimented slightly faster than standard HVJ.
Preparation of Equine Infectious Anemia Antigens for Diagnosis
Authors: Ueda Susumu , Samejima Togo , Kuroda Sadao , Nakamura Junji
Published: November 2013

The purification procedures and characteristics of an equine infectious anemia (EIA) antigen reacting specifically in the immunodiffusion test with sera from EIA‐infected horses were investigated. Partially purified antigens were extracted from both infected horse leukocyte cultures and the horse spleen principally by dialysis against 0.01m acetate buffer, pH 5.0. These extracts were further purified by filtering through a Sephadex G‐100 column. The purified antigens had a common antigenic determinant and were heat‐labile and soluble at pH 5.0. The results of immunoelectrophoresis and gel filtration suggested that the spleen‐derived antigen was of a smaller molecule than the culture‐derived antigen. The partially purified antigens from both sources were comparatively used for the detection of precipitating antibody in experimentally infected horses and in horses in the field. The two antigens showed an identical reaction with each serum tested. The two partially purified antigens were considered to be useful for diagnosis of EIA.
FP5 Factor, an Undescribed Sex Factor of Pseudomonas aeruginosa
Authors: Matsumoto Hideki , Tazaki Tadakatu
Published: November 2013

A Pseudomonas aeruginosa, strain Ps5–70 which was mercury‐resistant, was found to have a sex factor. This sex factor, designated as FP5 factor, mediated chromosome transfer by conjugation. Crosses between Ps5–70, FP5+ and PAO, FP2+ strain of Holloway were fertile, and Ps5–70 acted as the donor. Some of the recombinants between FP5+ and FP5– strains displayed maleness, and those recombinants usually acquired mercury resistance. So, transferability of FP5 factor itself and a very close association between the two determinants of FP5, one for fertility and the other for mercury resistance, were proved similar to the FP2 factor. Female sublines of Ps5–70 which behaved as recipients in intrastrain crosses were obtainable from cultures multiplied in the presence of acriflavin. Those females still retained mercury resistance. An R factor originated in P. aeruginosa interfered with the recombinant formation mediated by FP5 factor when the two episomes coexisted within a single cell.
Transformability of Group H Streptococcus Challis
Authors: Ito Takako , Hirano Toyo , Tomura Tsuneko , Yoshioka Morimasa
Published: November 2013

From a wild type strain Challis of the group H streptococcus, greening (Challis α) and β‐hemolytic (Challis β) colonies were isolated on horse blood agar. Both colonies formed greening on sheep blood agar, and no significant differences were found in their biological, serological and chemical analyses. They, however, showed clear differences on the transformability. Transformability, the producibility of competence‐provoking factor (CPF) and competency which have been reported on the Challis strain were all found in Challis β strain. On the other hand, Challis α strain did not produce antibiotic‐resistant transformants with the addition of CPF, and could not produce CPF even when the cells were cultured under various conditions of incubation or treated with lysozyme or detergents. The transformabilities of antibiotic‐resistant mutants obtained from the Challis β strain were lower than those of the original Challis β strain, as pointed out by other investigators, while the Challis α strain became transformable on antibiotic resistance only when it acquired streptomycin resistance. In the Challis β strain and the antibiotic‐resistant mutants of Challis α strain, the separate markers of streptomycin, penicillin, tetracycline, mitomycin C, as well as the combinations of these markers were found to be transformed at the highest rate in the strains having transformation of streptomycin resistance. The findings are discussed with respect to incorporation of deoxyribonucleic acid into recipient cells and to the reports of other workers.
A Generalized Transducing Phage of Serratia marcescens
Authors: Matsumoto Hideki , Tazaki Tadakatu , Hosogaya Shigemi
Published: November 2013

A temperate phage, phage PS20, which originated in a lysogenic strain of Serratia marcescens was established to be able to transduce various auxotrophic markers in S. marcescens strains at an equal frequency which ranged from 10–6 to 10–7 per infected cell. Electron microscopic observations show that phage PS20 has a head (550 A) hexagonal in outline and a tail (1000 A × 150 A) with a contractile sheath which is attached to the head.
Bacterial Function Can Contribute to Replication of Salmonella Phage P22
Authors: Yamagami Hiroshi , Yamamoto Nobuto
Published: November 2013

The UV sensitive bacterial mutants (abbreviated ms mutant) were isolated after N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (NG) mutagenesis. The ms mutants were found to be Hcr+, Rec+ and sensitive to UV and NG. The significance of the ms mutants was inability to support the multiplication of several mutant phages of P22 which were also isolated by NG mutagenesis. These characters of ms mutants were caused by a single point mutation. Some of the mutant phages unable to grow in ms host were also unable to grow in recA hosts and determined to be erf mutants. The remaining phages were classified in two groups. One group of mutants formed minute plaques on wild type host and fell in the same complementation group (x mutation) and the other formed larger plaques than wild type plaques and belonged to an another complementation group (y mutation). These newly found two genes were essentially involved in the replication of viable P22 DNA. Of the three mutants, x and erf were moderately sensitive to UV irradiation in comparison with y and wild type. Study of lysis patterns showed that the y or erf infected ms culture lysed despite their inability of forming viable progenies, whereas, the x infected ms culture did not lysed. The new x and y genes were both mapped in the early gene segment of P22 chromosome, on the left side of erf gene and between c3 and c2 genes, respectively. Based on these findings the contribution or substitution of the bacterial functions to the replication of bacteriophage was discussed.
The Effect of Rifampicin on Mycobacterium leprae in Mice
Authors: Matsuo Yoshiyasu
Published: November 2013

Rifampicin was shown to be highly active against Mycobacterium leprae in mice, completely inhibiting the in vivo multiplication of this organism at a dosage of 0.1 mg per day by gavage. When drug administration was started after the establishment of M. leprae in mice (30 weeks after infection), rifampicin stopped any further bacillary increase. A second inoculation of the organisms from treated mice showed complete loss of viability. Exposure of M. leprae to rifampicin for a short period before infection significantly reduced the viability of the organism.
Photochromogenesis
Authors: Koyama Yasumasa , Yazawa Yoshitaka , Yamagishi Saburo , Arai Tadashi
Published: November 2013

The effect of light on the pigmentation of various strains of Nocardia, Corynebacterium, Arthrobacter, Brevibacterium, and Flavobacterium was investigated. It was revealed that thirty out of fifty‐seven strains of Nocardia, two out of fifteen strains of Corynebacterium, two strains of Arthrobacter, six out of thirteen strains of Brevibacterium and two out of fourteen strains of Flavobacterium were photochromogenic; i. e., these strains produced yellow or orange pigments when grown under illumination but entirely unpigmented in total darkness. From these results, it may be concluded that photochromogenicity is not a particular phenomenon limited to specific species, but a common, widely distributed phenomenon in nonphotosynthetic bacteria.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Yamada Hiroyasu , Mashiba Harukazu , Takeya Kenji
Published: November 2013

C57BL/6 and AKR mice were treated with hamster erythrocytes (HRBC) in complete Freund's adjuvant (CFA) or incomplete Freund's adjuvant (IFA) and the development of delayed hypersensitivity and antibody production were examined. 1) Delayed hypersensitivity against HRBC antigen, as determined by the peritoneal macrophage disappearance test, was detected in mice sensitized with HRBC in CFA but not in those sensitized with HRBC in IFA. 2) Antibody production against HRBC or hapten TNP after a booster injection of HRBC or trinitrophenylated HRBC (TNP‐HRBC) in saline was enhanced by pretreatment with HRBC in CFA or IFA. 3) Delayed hypersensitivity was not detectable after a booster sensitization with HRBC in CFA in mice which had been pretreated with HRBC in IFA 2 weeks earlier. In the mice treated with both HRBC in IFA (day −21) and in CFA (day −7), however, an enhanced antibody production against HRBC or TNP was detected after an intravenous injection with HRBC or TNP‐HRBC in saline (day 0). These results suggest that sensitized effector lymphocytes in delayed hypersensitivity and helper cells in antibody production may be derived from the same pool of unprimed T cells. The pool of unprimed T cells with a capacity to differentiate into either type of primed T cells may be exhausted after pretreatment with the antigen in IFA, and the primed helper T cells may not be able to differentiate into sensitized lymphocytes even after sensitization with the antigen in CFA, which favors development of delayed hypersensitivity in normal controls.
Susceptibility of Mycoplasma pneumoniae to Antibiotics In Vitro
Authors: Niitu Yasutaka , Kubota Hideo , Hasegawa Sumio , Komatsu Shigeo , Horikawa Masahiro , Suetake Tomiko
Published: November 2013

The antibiotic susceptibility of laboratory strains, Mac and FH, and isolates of Mycoplasma pneumoniae was determined in broth media. With the Mac strain, the minimum inhibitory concentration (MIC) increased with the increase in inoculum size for most antibiotics and the final minimum inhibitory concentration as determined during a month's incubation did for all antibiotics. The antibiotics studied were classified by the MIC into three groups; the first group with the order of 1/100 μg/ml (erythromycin, josamycin, and leucomycin), the second with the order of 1/10 μg/ml (oleandomycin, tetracycline, and spiramycin), and the third with the order of 1 or 10 μg/ml (lincomycin, chloramphenicol, and streptomycin). With the FH strain, the MIC of streptomycin was much lower, showing that the Mac strain was resistant to streptomycin. The susceptibility of 152 isolates from 99 patients to erythromycin, josamycin, tetracycline, and chloramphenicol was essentially similar to that of the Mac strain except for two isolates, which had been isolated from a patient during erythromycin therapy and exhibited resistance to erythromycin and josamycin. The MIC of streptomycin was distributed at 1 μg/ml in 93% of the isolates and at 100 μg/ml or over in 6%. It was considered that streptomycin‐resistant strains occur in M. pneumoniae. Of nine patients who had been given erythromycin, tetracycline, chloramphenicol, or Clotaon for more than a week, the isolates after therapy showed decreased susceptibility to erythromycin in one patient and did not to the antibiotics given in the other eight patients.
Production of Mouse Interferon with High Titers in a Large‐Scale Suspension Culture System1
Authors: Sano Emiko , Matsui Yoshiko , Kobayashi Sigeyasu
Published: November 2013

L‐MS cells, adapted to grow in suspension, were obtained by selection from a high interferon (IF)‐producing line of mouse L cell monolayers. A large volume of L‐MS cells (20 liters or more; 1–2 × 1010 cells) was readily grown in a spinner culture, retaining their ability to produce high yields of IF in serum‐free medium following induction with Newcastle disease virus (NDV). The optimal condition for the production of IF in the suspension culture of L‐MS cells was established. The system also proved itself to be susceptible to IF induction by polyinosinic‐polycytidylic acid (Poly I · Poly C) and by NDV inactivated with ultraviolet light (NDV‐UV). By employing the present system, large quantities of mouse IF of a high titer could be routinely prepared.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Mashiba Harukazu , Sato Mikio , Takeya Kenji
Published: November 2013

Patterns of proliferation of antibody‐forming cells after an intravenous immunization with hamster erythrocytes (HRBC) were compared in groups of mice possessing different activities of thymus‐derived lymphocytes (T cells). 1) Marked differences in the numbers of hemolysin plaque‐forming cells (PFC) after HRBC injection were found among the low‐ and high‐responder normal mice and those pretreated with HRBC in complete Freund's adjuvant (CFA) or incomplete adjuvant (IFA), and they appeared to depend primarily upon the different rates of proliferation of antibody‐forming cells rather than on the numbers of antigen‐specific lymphocytes initiating the antibody response. 2) The numbers of hemolytic foci were slightly larger in mice with large numbers of PFC (normal SL mice, the pretreated SL and C57BL/6 mice) than in those with small numbers of PFC (normal C57BL/6 mice). The numbers of hemolytic foci increased at almost the same rate from day 2 to day 3 in both groups, while the numbers of PFC increased more efficiently in mice with large numbers of PFC than in those with small numbers of PFC from day 2 to day 3. Individual hemolytic foci appeared to contain larger numbers of PFC in mice with large total numbers of PFC than in those with small total numbers of PFC. 3) The numbers of rosette‐forming cells (RFC) were increased by pretreatment with HRBC in CFA and by pretreatment with HRBC in IFA to almost the same extent. Rates of increases in PFC were, however, larger by pretreatment with HRBC in CFA than with HRBC in IFA. These results suggested that the activity of the T cell determined not only the rates of proliferation of antibody‐forming cells but also the antibody‐producing capacity of each cell.
A Comparative Study of Mycobacteria from Patients' Room Dusts and from Sputa of Tuberculous Patients
Authors: Tsukamura Michio , Mizuno Shoji , Murata Hiroshi , Nemoto Hisashi , Yugi Hiroyuki
Published: November 2013

The source of mycobacteria other than Mycobacterium tuberculosis occurring in sputa of tuberculous patients as casual isolates was investigated by comparing the occurrence rate of mycobacterial species between patient's room dust and patient's sputum. Almost all species of mycobacteria recovered from sputa could be found in dusts obtained from the rooms. However, the percentage of occurrence of the mycobacterial species in dusts differed from that in sputa. In dusts, Mycobacterium fortuitum (39.6%), Mycobacterium nonchromogenicum (23.5%) and Mycobacterium gordonae (16.7%) occurred in high frequencies, whereas Mycobacterium intracellulare (69.6%), M. gordonae (5.9%), Mycobacterium scrofulaceum (5.2%), and Gordona bronchialis (10.4%) were the main species found in sputa. The patterns of occurrence of mycobacterial species as illustrated above may suggest that pathogenic mycobacteria survive in the respiratory tract while nonpathogenic ones are destroyed there, thus the human body acts as a selective medium for the pathogenic mycobacterial species. Serotype studies on strains of M. intracellular as casual isolates indicated that those from sputa of tuberculous patients were different from those derived from patients with lung diseases due to this species of mycobacteria. These results led us to the conclusion that mycobacteria in patients' room dusts were the source of mycobacteria occurring in sputa as casual isolates, particularly the pathogenic mycobacteria in dusts are more likely to survive in the human respiratory tract, occasionally multiplying and causing disease under favorable circumstances.
Studies of Mycobacterium lepraemurium in Cell Culture
Authors: Matsuo Yoshiyasu
Published: November 2013

A serial increase in the number of Mycobacterium lepraemurium with successful subcultures has been obtained in the mouse foot pad (MFP) cell culture. Special attention has been given to maintaining the infected cells for longer periods; 1) the infected cells were incubated at 30 C rather than at 37 C, and 2) the concentration of serum in the culture medium was reduced from 10 to 2% as soon as a monolayer growth of the transferred cells was obtained. There have been cumulative bacterial increases of 1.47 × 1017 and 1.84 × 1015 fold for the Kurume‐42 strain during a period of 1255 days, and 2.23 × 109 and 3.89 × 105 fold for the Hawaiian strain during periods of 831 and 572 days. The overall generation times were estimated at 22.0, 24.8, 26.8, and 30.8 days, respectively. All attempts to grow the acid‐fast bacilli obtained in cell cultures on artificial culture media have failed. The ability of the organisms to produce typical lesions in mice has been well preserved.
Nitroblue Tetrazolium (NBT)‐Dye Test and the Myeloperoxidase Reaction of Human Leukocytes
Authors: Okuda Kenji , Tadokoro Ichiro , Noguchi Yoshikuni , Okuda Kyoko
Published: November 2013

Nitroblue tetrazolium (NBT)‐dye tests were performed on blood specimens obtained at the time of admission or during the course of hospitalization from 498 patients and 150 healthy persons. In the healthy group, persons over 3 years of age showed mean percentages of 19.16 ± 5.03 of NBT‐positive neutrophils. On the other hand, infants from 3 months to 3 years of age showed decreased values of NBT‐dye reduction. Patients suffering from acute bacterial or fungal infections, or cryoglobulinemia showed high percentages of NBT‐positive neutrophils. Most patients with viral or tuberculous infections demonstrated normal or slightly attenuated values of NBT‐dye reduction. Patients who had been receiving corticosteroids for long periods, those in severe stages of diabetes mellitus, and those receiving radiotherapy or anticancer drugs had remarkably impaired NBT‐dye reduction. We examined the myeloperoxidase reactions, since we knew that patients with myeloperoxidase deficiencies have been reported to have disseminated candidiasis and to be defective in the ability of neutrophils to kill Candida albicans. We confirmed the fact that most persons had equal levels of myeloperoxidase and were unable to find anyone with a deficient myeloperoxidase syndrome among 450 individuals.
Factors Influencing the Establishment of Persistent Infection of HVJ (Sendai Virus) in L Cells
Authors: Sugamura Kazuo , Tozawa Hideki , Homma Morio , Ishida Nakao
Published: November 2013

Infection of a subline of L cells adapted to grow in suspension (Ls) with Fushimi strain of HVJ (HVJ‐F) resulted in a virus carrier state. Ls cells, when cultured in monolayer, showed morphological changes following infection of HVJ‐F and were detached from the glass wall. However, when the detached cells were transferred to a new environment of suspension culture within 5 days after infection, the carrier state was again established. HVJ‐F caused only lethal infection in L cells maintained exclusively in monolayer (Lm). On the other hand, both Ls and Lm, irrespective of their culture conditions, were lethally infected by Nagoya 1–60 strain of HVJ. The overall results showed that culture condition as well as the kind of host cells or virus strains is an important factor regulating the establishment and maintenance of the virus carrier state.
Quantitative and Qualitative Analyses of the Antibody Response of Adult Mice Tolerized with Deaggregated Bovine γ‐Globulin
Authors: Muramatsu Shigeru , Hosono Masamichi
Published: November 2013

The antibody response of mice to bovine γ‐globulin(BGG) was suppressed either specifically by an intravenous injection of deaggregated soluble BGG (sBGG) or nonspecifically by X‐irradiation. Immunization with the subcutaneous injection of BGG in Freund's incomplete adjuvant was given to mice either various days after sBGG injection or immediately after X‐irradiation. Antigen‐elimination (AE) test and passive hemagglutination(PHA) test were employed for estimating the immune status. The AE test indicated that the induction of tolerance was accomplished in the first 2 days after sBGG injection and that the tolerant state was stable at least for about 30 days thereafter. The degree of suppression by 1000 μg of sBGG corresponded to that obtained by X‐irradiation at the dose of 400 R or more, and 100 μg of sBGG was equivalent to 300 R X‐irradiation. The PHA test indicated, however, that such a correspondence as mentioned above between the dose of tolerogen and that of X‐irradiation was not so stable as was seen by the AE test. Thus, the PHA titers of tolerized animals tended to recover up to the level of untolerized animals during the period of time from 10 days to 20 days after the tolerogen injection. Such discrepancies between the features in the AE test and those in the PHA test seemed attributable to a low avidity antibody formation in the tolerized animals, as judged by the hemagglutination‐dissociation test. Hemagglutination by means of the sera from tolerized animals was seen to be reversed by the addition of free antigen more easily than the hemagglutination achieved by the sera of control animals or X‐irradiated animals. The relationship between PHA titers and AE capacities of antibodies was investigated by the passive immunization of normal mice previously given the antigen. The result showed that the PHA titer did not always correlate with the AE capacity.
Studies on the Neutralization of Herpes Simplex Virus
Authors: Ide Kazuko , Yoshino Kamesaburo
Published: November 2013

Millipore‐filtered herpes virus and a hyperimmune rabbit serum were reacted to analyze the unneutralizable persistent fraction (PF). When the PF was serially diluted with antibody‐containing diluent, the plaque‐formers reduced in number. When the serial dilutions were made with antibody‐free diluent, plaque numbers were disproportionately smaller in lower dilutions. The PF filtered through a 0.22μ Millipore membrane showed only a slight loss of infectivity, but a further incubation of the filtrate at 37 C resulted in a marked reduction of titer. This effect was less pronounced when the membrane porosity was larger. Additional virus given to the PF was quickly neutralized by excess antibody. On the other hand, dilution of the virus–serum mixture followed by incubation at 37 C or sonication did not further reactivate the virus. When neutral complexes were sedimented by ultra‐centrifugation and resuspended in antibody‐free diluent, a partial reactivation slightly exceeding the usual PF level occurred with a concomitant release of antibody. It is proposed that the PF may be free virus resulting from reversible virus‐antibody reaction.
Ionic Germination of Spores of Clostridium perfringens Type A
Authors: Ando Yoshiaki
Published: November 2013

Spores of Clostridium perfringens NCTC 8238 underwent so‐called “ionic germination” under suitable conditions in aqueous solutions of either inorganic or organic salts. Chlorides of alkali metal ions except for Na+ were the most effective in inducing germination. Chlorides of alkali earth metal ions were effective to some extent, but those of heavy metal ions were rather inactive. Potassium salts of halogens, nitrogen, sulfur and phosphorus, as well as organic acids including fatty acids and polycarboxylic acids were also germinative. Preheating the spores at 80 C for 10 min was essential for rapid and complete germination by these salts. Maximum germination occurred at an incubation temperature of 40 C and at a fairly wide range of pH values (5.0–9.0). The germination was markedly inhibited by various uncoupling reagents such as carbonylcyanide m‐chlorophenylhydrazone, pentachlorophenol, hexachlorophene (HCP), and dicumarol. This inhibition was reversible because removal of these reagents allowed germination to proceed. The inhibiting effect of HCP was highly dependent on pH, suggesting that a molecular dissociation of the reagent affects the inhibition. Furthermore, the inhibition of germination by HCP was competitively overcome at higher KCl concentrations. The possible mechanism of ionic germination and the inhibitory effect of uncouplers on the ionic germination are discussed.
Antigenic Analyses of Sequential Isolates of Coxsackie A16 Virus from 1964 to 1970 in Japan
Authors: Fujimiya Yoshiaki , Suto Tsunehisa , Morita Morihiro , Ishida Nakao
Published: November 2013

Biological and serological comparisons of sequential isolates of Coxsackie A16 virus (Cox A16) in 1951 through 1970 were made to elucidate the mode of appearance of the mutant which caused a nationwide epidemic of hand‐foot‐and‐mouth disease (HFMD) in 1970. Most strains isolated from feces of healthy children in 1964, 1966 and 1967 were serologically similar to the prototype strain isolated in 1951. However, some strains isolated from these healthy children in 1966 and 1967 were serologically and biologically related more closely to the epidemic strain isolated in 1970. Thus sequential isolates of Cox A16 could be roughly classified into two groups, i.e., the prototype strain‐like and the epidemic strain‐like groups.
Regular Array in the Cell Wall of Lactobacillus fermenti as Revealed by Freeze‐Etching and Negative Staining1
Authors: Kawata Tomio , Masuda Kuniyoshi , Yoshino Kazuhiro , Fujimoto Michimasa
Published: November 2013

Freeze‐etching of Lactobacillus fermenti F‐4 (NCTC 7230) revealed that the outer layer of the cell wall was composed of a regular array in which parallel lines ran obliquely to the longitudinal axis of the cell with an average distance between the centers of about 9.6 nm and were intersected by thinner lines with an average periodicity of approximately 6.2 nm at an angle of about 75°. Occasionally the direction of the striation was discontinuously shifted near one end of the cell. Beneath the regular array the middle cell wall layer packed with granules and the smooth inner cell wall layer were discernible and the mesosomes were also visible in the cytoplasm. When the ultrastructure of isolated outer cell wall fragments was examined by negative staining, the regular array appeared to be composed of subunits, about 3.6 nm in diameter, which were arranged in a tetragonal pattern. The tetragonal array consisted of the subunits in rows in two directions at an angle of about 75° to each other. The average spacing between the rows was about 9.3 nm in one direction and 5.5 nm in the other direction.
Electron Microscopic Observations of Lipid Antigens With Special Reference to the Wassermann Antigen
Authors: Kanemasa Yasuhiro
Published: November 2013

The Wassermann antigen is a lipid micelle, and the essential component of the hapten is cardiolipin. It requires the addition of lecithin and cholesterol as auxiliary lipids. We conducted a study on the role of auxiliary lipids in the formation of the micelle structure using negative staining and observing with the electron microscope. The following results were obtained: (1) Cardiolipin, when dispersed in Mg–NaCl solution, presented an irregular network structure. Lecithin, on the other hand, revealed a regular lamellar structure. Cholesterol, since it is nonpolar, immediately agglutinated and precipitated when suspended in the aqueous system. (2) A combination of cardiolipin and lecithin presented a regular lamellar structure, but no network structure of cardiolipin was observed in the visible field. The combination of cardiolipin and cholesterol revealed cardiolipin surrounding structureless cholesterol with its network structure. The combination of lecithin and cholesterol exhibited a lamellar structure surrounding the cholesterol. (3) A combination of all three revealed a variegated picture of a lamellar structure formed by cardiolipin and lecithin surrounding the cholesterol as a core or of onionskin‐like micelles attached to the above structure.
Enhanced Production of Interferon by Temperature Shift‐Down from 37 C to 25 C in Rabbit Cell Cultures Stimulated with Newcastle Disease Virus
Authors: Kojima Yasuhiko , Yoshida Fumio
Published: November 2013

Preincubation of Newcastle disease virus (NDV)‐infected RK13 cells at 37 C considerably enhanced subsequent interferon (IF) production at 25 C. The optimal length of this preincubation was about 7 to 9 hr. It was postulated that the intracellular events leading to IF production in NDV‐induced cells might proceed in two distinct phases, early and late; the early phase to prepare the cells for IF synthesis proceeds only at 37 C whereas the late phase in which IF is readily synthesized occurs both at 37 C and at 25 C. Similar events seemed to occur in cultures of cells from in vivo induced rabbits. This concept was substantiated by experiments using actinomycin D and cycloheximide, which suggested that protein synthesis at 37 C during the first 7 hr was essential for subsequent IF production at 25 C. A prolonged preincubation at 37 C resulted in decreased IF production, suggesting that a putative regulatory protein might be synthesized at 37 C which could not be readily synthesized at 25 C.