bluuebunny/crossref_metadata_2025 huggingface dataset:
Delayed‐Type Hypersensitivity (DTH) Response to Dengue Virus in Mice: Effect of Route of Sensitization and Splenectomy
Authors: Pang Tikki , Yap Kok Leong , Devi Shamala , Wong Phooi Yeen
Published: November 2013

This study was designed to determine the role of the sensitization route and the spleen in the development of delayed‐type hypersensitivity (DTH) to dengue virus in mice. DTH was measured by footpad swelling response. Strong but transient DTH was produced in cyclophosphamide (CY) pretreated mice sensitized subcutaneously (s.c.) or intravenously (i.v.) with dengue virus type 4. Subcutaneous inoculation of virus in incomplete Freund's adjuvant (IFA) further enhanced the DTH elicited. The time course of DTH generated by s.c. and i.v. sensitization were similar with the peak reactivity seen on day 6 after sensitization. Poor DTH was observed in mice given an i.p. inoculation even when CY and/or IFA were used. Intracerebral (i.c.) inoculation also sensitized mice poorly. Splenectomized mice showed enhanced DTH response when compared to intact mice. In contrast to intact mice, pretreatment of splenectomized mice with CY did not alter the DTH level. Splenectomized mice inoculated s.c. with virus in IFA showed poorer DTH than mice sensitized with virus alone.
Effect of Membrane Fluidizers on the Number and Affinity of Chemotactic Factor Receptors on Human Polymorphonuclear Leukocytes
Authors: Tomonaga Akimitsu , Hirota Masaki , Snyderman Ralph
Published: November 2013

Chemotaxis by leukocytes appears to be initiated by the binding of chemoattractants to specific cell surface receptors. In other biological systems, the affinity and functional activity of membrane receptors are regulated by the local microviscosity. The present studies were undertaken to determine if the number and/or affinity of chemotactic factor receptors expressed on human polymorphonuclear leukocytes were similarly affected. Aliphatic alcohols and cis‐vaccenic acid, agents known to decrease membrane microviscosity, were studied for their effects on the binding of the radiolabeled chemoattractant f‐Met‐Leu‐[3H]Phe to human polymorphonuclear leukocytes. Butanol and propanol increased the number of f‐Met‐Leu‐[3H]Phe binding sites approximately 1.5 fold. More dramatically, these same agents enhanced the affinity of the receptor by ten‐fold, without affecting the specificity of the receptor. Similarly, cis‐vaccenic acid enhanced both the number and affinity of this chemotactic factor receptor on human polymorphonuclear leukocytes contain cryptic receptors for the N‐formylated peptide chemotactic factors, but more importantly that the affinity of these receptors can exist in more than one state and can be modulated by membrane microviscosity. Alterations of membrane fluidity in leukocytes during chemotaxis may be an important mechanism for regulating their sensitivity to chemoattractants.
Further Isolation of a Recombinant Virus (H1N2, Formerly Hsw1N2) from a Pig in Japan in 1980
Authors: Yasuhara Hisao , Hirahara Tadashi , Nakai Masahisa , Sasaki Norimasa , Kato Jiro , Watanabe Takayoshi , Morikawa Masamichi
Published: November 2013

In September 1980, an outbreak of febrile respiratory disease was observed in a herd of sows (1‐2 years of age) in Ehime Prefecture, Japan. Most of the swine showed clinical signs of disease such as depression, anorexia, fever, nasal discharge, and cough.

A hemagglutinating agent was isolated from a nasal swab from one of the diseased pigs. By cross‐hemagglutination‐inhibition and neuraminidase‐inhibition tests with antisera to influenza viruses of swine origin, the isolate was identified as an influenza A virus of the H1N2 (former designation, Hsw1N2) subtype, and designated A/swine/Ehime/1/80 (H1N2).

Significant antibody rises against the surface antigens of the isolate were found in convalescent swine sera.

The distribution of antibody against H1N2 virus in swine sera in Ehime Prefecture was examined. Seven (8%) of 93 sera collected after the outbreak (in 1981) showed antibodies to only H1 and N2 antigens but none of the sera before the outbreak contained such antibodies, indicating that H1N2 virus had been restrictedly prevalent among swine but was not wide‐spread until 1981.
Characterization of Monoclonal Antibodies against Etiological Agents of Weil's Disease
Authors: Kobayashi Yuzuru , Tamai Tomonori , Oyama Takashi , Hasegawa Hitoshi , Sada Eiji , Kusaba Tomohiro , Hamaji Masaharu
Published: November 2013

Monoclonal antibodies against etiological agents of Weil's disease were produced by cell fusion technology. Twenty hybridomas were produced through the fusion of P3×63Ag8.653 cells with spleen cells from BALB/c mice immunized against Leptospira interrogans serovar icterohaemorrhagiae RGA strain and serovar copenhageni Shiromizu and M20 strains. Reactivities of the antibodies produced by the hybridomas were determined by the microscopic agglutination test. Among the five hybridoma antibodies to the RGA strain, two reacted specifically to serovar icterohaemorrhagiae, two reacted to serovar icterohaemorrhagiae at a high titer and serovar copenhageni at a low titer, and one reacted to serovars icterohaemorrhagiae, copenhageni, pyrogenes, and canicola. Of the ten hybridoma antibodies to the Shiromizu strain, one reacted specifically to serovar copenhageni, seven reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, and two exhibited intermediate properties. Of the five hybridoma antibodies to the M20 strain, three reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, one reacted to serovar copenhageni at a low titer and serovar icterohaemorrhagiae at a high titer, and one reacted to serovars copenhageni, icterohaemorrhagiae, and pyrogenes. The results revealed that each serovar has its own antigen(s) and their common antigens. In addition, 20 strains of leptospires were recently isolated and tested with three monoclonal antibodies characterized by different reactivities. Twenty strains were clearly identified by their antibodies, i.e., 16 strains were identified as serovar icterohaemorrhagiae and three strains were identified as serovar copenhageni. The remaining strain, which was not agglutinated by three antibodies, was identified as serovar autumnalis by an agglutination test with immune rabbit sera.
Fluctuation of Antiviral Resistance in the Intestinal Tracts of Nude Mice Infected with a Mouse Adenovirus
Authors: Umehara Keiko , Hirakawa Minako , Hashimoto Kazuo
Published: November 2013

Oral administration of adenovirus strain K87 to BALB/c nude mice resulted in viral proliferation in the intestinal tract up to around week 6 at which point replication was suppressed. In other words, the host acquired resistance. However, this resistance was temporary and the viral infection persisted over a long period with repeated periods of proliferation and resistance. That the appearance of this resistance is the result of infecting mice with the virus and is not due to age difference per se was made clear through experimentation with nude mice of different age groups. However, it was indicated that increase in age is involved in the decreased rate of reproliferation following initial suppression. No evidence of the virus was obtained from any other organ throughout the infection. Furthermore, throughout the persistent infection, even during the aforementioned periods of resistance, no neutralizing antibody was detected from sera, intestinal wall or intestinal content. When spleen cells from BALB/c heterozygous littermate mice was transferred to the nude mice, an earlier onset of antiviral resistance was seen than in nude mice without the transfer, and this was accompanied by a rise in neutralizing antibody titer. From these results, it is believed that the resistance characteristic of nude mice infected by mouse adenovirus is dependent on some factor other than the neutralizing antibody invoked resistance exhibited by euthymic mice.
Isolation of Rickettsia tsutsugamushi Antigenically Different from Kato, Karp, and Gilliam Strains from Patients
Authors: Tamura Akira , Takahashi Kumiko , Tsuruhara Takashi , Urakami Hiroshi , Miyamura Sadao , Sekikawa Hiroho , Kenmotsu Minora , Shibata Mitsuo , Abe Shoya , Nezu Hiroko
Published: November 2013

Rickettsia tsutsugamushi strains from three recent patients of Tsutsugamushi disease in Niigata Prefecture were isolated primarily in mice and then in L cell cultures. By this procedure, low virulent strains to mice, as well as high virulent ones, could be isolated and cultivated serially in L cell cultures, suggesting the usefulness of L cells for isolation of this species of rickettsia. Each newly isolated strain was identified as a member of R. tsutsugamushi from the results of cross immunological tests and morphological observation. On the other hand, it was recognized that one of these rickettsiae showed immunological properties distinguishable from the prototype strains of Kato, Karp, and Gilliam by the cross complement fixation test, and also had low virulence in mice.
Effect of Clostridium perfringens Beta Toxin on Blood Pressure of Rats
Authors: Sakurai Jun , Fujii Yoshio , Dezaki Keiko , Endo Koichi
Published: November 2013

Guanethidine treatment or adrenal medullectomy significantly inhibited the elevation in blood pressure induced by Clostridium perfringens beta toxin, and the combination of the two drastically reduced the pressure rise, to less than 19% of that in control rats. When rats were pretreated with tetrodotoxin or hexamethonium, the toxin‐evoked rise was significantly inhibited. Elevation in blood pressure induced by the toxin in spinal rats tended to be less than that in control rats. When investigated by a microscopical technique, arteriolar constriction in the mesenteric vasculature was observed after the blood pressure elevation induced by the toxin reached a maximum. Blood flow in the skin decreased with an increase in blood pressure following intravenous injection of the toxin. It is concluded that beta toxin acts on the autonomic nervous system and produces arterial constriction.
Enhancement of Newcastle Disease Virus‐Induced Fusion of Mouse L Cells by Sodium Vanadate
Authors: Yamamoto Fumi , Fujioka Hisashi , Iinuma Masao , Takano Michiko , Maeno Koichiro , Nagai Yoshiyuki , Ito Yasuhiko
Published: November 2013

Sodium vanadate enhanced Newcastle disease virus (NDV)‐induced cell fusion in L cells, and there was a direct correlation between the degree of cell fusion and the dose of vanadate added. When anti‐F protein of NDV monospecific antiserum was added to the culture fluid of L cells infected with NDV, the enhancement of cell fusion was suppressed. In contrast, neither anti‐HN nor anti‐M protein monospecific antiserum inhibited the enhancement. Incubation at low temperature (4 C) and addition of sodium azide to the culture fluid suppressed the enhancement. The suppression by azide was seen only when the drug was added within 5 min after the beginning of incubation of NDV‐infected L cells with vanadate. On the other hand, incubation at low temperature inhibited the enhancement at any time during incubation with vanadate. Cytochalasin D also inhibited the enhancement if it was added at any time during incubation with vanadate.
Cultured Human Monocytes, Granulocytes and a Monoblastoid Cell Line (THP‐1) Synthesize and Secrete Immunosuppressive Acidic Protein (a Type of α1‐Acid Glycoprotein)
Authors: Shibata Yoshimi , Tamura Keiji , Ishida Nakao
Published: November 2013

Immunosuppressive acidic protein (IAP, pI 3.0) is a type of α1‐acid glycoprotein (α1‐AG), The secretion of IAP into the culture fluids of different sub‐populations of human peripheral blood leukocytes was examined by a newly devised passive hemagglutination (PHA)‐inhibition test. Human peripheral monocytes, an established monoblastoid cell line (THP‐1) and peripheral granulocytes produced IAP. However, neither T nor B lymphocytes, nor lymphoblastoid cell lines induced by TCGF or EB virus respectively, produced IAP. The IAP concentration reached a maximum (215 ng/ml) in the culture fluids of peripheral monocytes (1 × 106/ml) when monocytes were stimulated by the addition of either immune complex, carrageenan or endotoxin.

The synthesis de novo and shedding of IAP by THP‐1 were demonstrated by the immunoprecipitation of radioactive IAP in the culture fluids of [3H]leucine‐labeled cells. SDS‐polyacrylamide gel electrophoresis of the immunoprecipitates showed two peaks of radioactivity, one comigrated with authentic IAP at 50,000 daltons, and the other at 38,000 daltons, suggesting that two different forms of IAP (and/or α1‐AG) are produced from human monocytes.
Detection of Fine Spiral Structures (Spirosomes) by Weak Sonication in Some Bacterial Strains
Authors: Matayoshi Seiken , Oda Hiroshi
Published: November 2013

Fine spiral structures (spirosomes) were observed in cell suspensions of five species of bacteria just after weak sonication. The structure is morphologically indistinguishable from the spirosome reported for Lactobacillus species. The molecular weight of the protein of the spirosomes from nine strains was about 94,000 to 95,000 as determined by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The difference in the molecular weight among these spirosomes was not very great, but there were slight differences among the strains from which the spirosomes were derived.
Activation of Human Monocytes in Leprosy
Authors: Ohkawa Susumu
Published: November 2013

In leprosy, the common etiologic agent is the same Mycobacterium leprae, but the clinical manifestations are various, including the tuberculoid and lepromatous types. In tuberculoid type leprosy, macrophages in the granuloma differentiate into epithelioid cells; in the lepromatous type, in contrast, they differentiate into lepra cells containing multiple M. leprae. Thus host factors, which regulate macrophage activities, determine the type of leprosy.

To understand such regulation of macrophage activities, we assayed superoxide production, hydrogen peroxide production and glucose consumption in monocytes in vitro.

Glucose consumption spontaneously increased, with lymphokine enhancing the consumption rate. Superoxide production increased spontaneously and decreased from the 4th day; lymphokine added on the 4th day supressed the decrease of superoxide production. Hydrogen peroxide production increased until the 3rd day of culture. Twenty‐four hour incubation with lymphokine, from day 0 to the 1st day, had no effect on hydrogen peroxide production, while from the 2nd to 3rd day it enhanced such production. Supernatants of lymphocytes incubated with M. leprae were prepared from tuberculoid and lepromatous patients. Tuberculoid supernatant enhanced reactive oxygen production and glucose consumption, while that from lepromatous patients had no remarkable effect on glucose consumption or reactive oxygen production. The range of spontaneous increase and decrease of reactive oxygen production was greater than the regulatory effect of lymphokine on these activities.

These data show that rapid provision of new monocytes to the granuloma is one of the important factors in the defense mechanism, that lymphocytes separated from lepromatous patients are not activated in response to M. leprae antigen, and that they do not secrete corresponding lymphokines.
Heat Resistance, Spore Germination, and Enterotoxigenicity of Clostridium perfringens
Authors: Ando Yoshiaki , Tsuzuki Toshibumi , Sunagawa Hiroyuki , Oka Shigemi
Published: November 2013

Heat resistance at 95 C, heat activation at 75 C, and germination response were determined for spores of 10 serotype strains of Clostridium perfringens type A, including five heat‐resistant and five heat‐sensitive strains. The D95‐values ranged from 17.6 to 63.0 and from 1.3 to 2.8 for the heat‐resistant and the heat‐sensitive strains, respectively. The heat‐activation values, the ratios between the heated and unheated viable counts of spore suspensions, ranged from 0.0035 to 0.65 and from 6.5 to 60.0 for the heat‐sensitive and the heat‐resistant strains, respectively. Spores of these strains were divided into two distinct germination types on the basis of their germination response; spores of the heat‐resistant strains germinated in KCl medium after heat activation (K‐type), and spores of the heat‐sensitive strains germinated in a mixture of L‐alanine, inosine, and CaCl2 in the presence of CO2 without heat activation (A‐type). The strains were tested for enterotoxigenicity by a reversed passive latex‐agglutination (RPLA) test. All the heat‐resistant strains were RPLA‐positive, whereas the heat‐sensitive strains were all RPLA‐negative. A total of 37 strains of the organism isolated from food‐poisoning outbreaks were tested for spore germination and enterotoxin formation. All of the 20 heat‐resistant strains showed K‐type spore germination and, except for three strains, were RPLA‐positive, whereas all of the 17 heat‐sensitive strains showed A‐type spore germination and, except for only one strain, were RPLA‐negative.
Comparison of Dengue Virus Plaque Reduction Neutralization by Macro and ‘Semi‐Micro’ Methods in LLC‐MK2 Cells
Authors: Morens David M. , Halstead Scott B. , Larsen Linda K.
Published: November 2013

A simplified ‘Semi‐Micro’ plaque reduction neutralization test (PRNT) for dengue antibody in LLC‐MK2 cells in disposable tissue culture plates is described. The assay compares favorably with the standard PRNT in glass prescription bottles, with relative sensitivity and specificity both 100% at a 1: 40 screening dilution by 70% plaque reduction criteria. The assay is easy to perform, economical of time, expense, and storage space, and is suitable for study of sera available in small volumes, such as those obtained on filter paper or by the capillary method. The LLC‐MK2 semi‐micro PRNT is an acceptable alternative to the standard PRNT, particularly in laboratories that use these cells routinely for other tissue culture work and for flavivirus vaccine development.
Characterization of Extracellular Substance of Vibrio anguillarum Toxic for Rainbow Trout and Mice
Authors: Kodama Hiroshi , Moustafa Mohamed , Mikami Takeshi , Izawa Hisao
Published: November 2013

An extracellular toxic substance was separated from the cell‐free culture filtrate of Vibrio anguillarum (strain NCMB571). Two fractions (GI and GII + III) obtained by Sephadex G‐200 chromatography following DEAE‐cellulose chromatography were lethal to rainbow trout and mice. Material separated from the GI fraction by Sepharose 4B affinity chromatography (GI‐A fraction) was lethal to these animals. By sodium dodecylsulfate Polyacrylamide gel electrophoresis, the GI and GI‐A fractions were found to be composed of components with molecular weights of 44K and 34K, and 44K, respectively. The 44K protein band was associated with carbohydrate. Peripheral vascular disorder was observed in fish and mice that died after inoculation with GI or GI‐A fraction. The toxic substance was sensitive to potassium periodate but was resistant to trypsin and acetone. Heat inactivation of the toxic substance was almost complete at 100 C for 20 min and complete at 121 C for 20 min. The toxic activity was not associated with hemolytic or proteolytic activity. Homologous antitoxin completely neutralized the toxic activity.
Neurovirulence in Mice of Neural Cell‐Adapted Canine Distemper Virus
Authors: Morikawa Yuko , Matsubara Yutaka , Yoshikawa Yasuhiro , Kai Chieko , Yamanouchi Kazuya
Published: November 2013

Neurovirulence of the Onderstepoort strain of canine distemper virus (CDV) adapted to human neural cell lines was determined by the intracerebral inoculation of DDD mice at 3 and 5 weeks of age. Intensity of neurovirulence was estimated by histopathological changes in the central nervous system and clinical symptoms. The original virus propagated in Vero cells induced leptomeningoencephalitis, whereas neuroblastoma‐adapted virus induced nerve cell degeneration and mild encephalitis with relatively low morbidity and fatality. In contrast, the viruses adapted to glioblastoma and oligodendroglioma caused high morbidity and fatality. The latter two viruses induced necrotizing encephalopathy including edema and hyperemia. In addition, the glioblastoma‐adapted virus induced formation of giant cells. The oligodendroglioma‐adapted virus caused demyelination and spongy state associated with degeneration of glial cells and axons. These observations are discussed in regard to a possible correlation between the neurovirulence of CDV in mice and its tropism for neural cells in vitro.
————‐Immunological Properties of Rickettsia tsutsugamushi Kawasaki Strain, Isolated from a Patient in Kyushu
Authors: Yamamoto Seigo , Kawabata Norihiko , Tamura Akira , Urakami Hiroshi , Ohashi Norio , Murata Michisato , Yoshida Yoshiya , Kawamura Akiyoshi
Published: November 2013

Nine isolates of Rickettsia tsutsugamushi were obtained from patients with Tsutsugamushi disease (scrub typhus) in Miyazaki Prefecture in Kyushu. Immunological analyses of these patients' sera and the isolates were performed by indirect immunofluorescence, indirect immunoperoxidase or immunoblotting techniques. In the analysis of reactions of the patients' sera with the prototype strains Gilliam, Karp, and Kato and with the isolates, sera from two patients, including Kawasaki, showed similar profiles and cross‐reaction with the two isolates recovered from the corresponding patients, but reacted only weakly with the prototype strains. With guinea pig polyclonal antibodies against the isolate and prototype strains, Kawasaki strain showed some degree of cross‐reaction with Gilliam strain but not with either Karp or Kato strain, nor with Shimokoshi strain which is known to be different antigenically from the prototype strains. Additionally, strain‐specific murine monoclonal antibodies against Gilliam, Karp, and Kato strains did not react at all with Kawasaki strain. These results suggest that the Kawasaki strain may be different antigenically from the prototype strains and Shimokoshi strain. The finding two strains of the same antigenic type (Kawasaki) among only nine isolates suggests the presence of Kawasaki‐type rickettsiae in Miyazaki Prefecture. Shimokoshi strain also did not react with these strain‐specific monoclonal antibodies, suggesting that strains of R. tsutsugamushi antigenically distinct from the prototype strains, such as Kawasaki and Shimokoshi strains, may easily be recognized by their non‐reactivity with these monoclonal antibodies.
No Involvement of Bovine Leukemia Virus in Sporadic Bovine Lymphosarcoma
Authors: Ogawa Yasuki , Sagata Noriyuki , Tsuzuku‐Kawamura Junko , Onuma Misao , Izawa Hisao , Ikawa Yoji
Published: November 2013

Using various portions of a molecularly cloned bovine leukemia virus (BLV) DNA as probes, the possible integration of a BLV genome or a BLV‐related sequence into the chromosomal DNA of sporadic bovine leukosis (SBL) tumor cells was investigated by Southern blotting analysis. Under stringent as well as non‐stringent conditions of hybridization, neither BLV nor BLV‐related sequence specific to SBL DNAs was detected in any SBL tumor examined. These results provide conclusive evidence for lack of the relation of BLV or a BLV‐related agent to SBL.
Monoclonal Antibodies against Chlamydia psittaci
Authors: Toyofuku Hajime , Takashima Ikuo , Arikawa Jiro , Hashimoto Nobuo
Published: November 2013

Five monoclonal antibodies were prepared against Chlamydia (C.) psittaci strain Pigeon‐1041 isolated from a feral pigeon in Sapporo. Reactions of these antibodies to chlamydiae were examined using five strains of C. psittaci and two strains of C. trachomatis in an enzyme‐linked immunosorbent assay, microimmunofluorescent test and complement fixation test. The antibodies were divided into two groups: three genus‐specific (A2, D2, and I21) and two strain‐specific (F2 and H9) antibodies. The antigenic determinant site of A2 was KIO4 sensitive, but those of D2, F2, and H9 were not affected greatly by KIO4 treatment. Nine C. psittaci strains from feral pigeons and 16 strains from budgerigars were classified into three groups and four groups, respectively, by reaction patterns against the monoclonal antibodies.
In vitro Intestinal Microflora‐Mediated Metabolism of Biliary Metabolites from 1‐Nitropyrene‐Treated Rats
Authors: Kinouchi Takemi , Nishifuji Keiko , Ohnishi Yoshinari
Published: November 2013

To investigate the modifying role of the intestinal microflora in the metabolism of 1‐nitropyrene (1‐NP) via enterohepatic circulation, we collected bile from male Wistar rats administered [3H]1‐NP orally. The bile was mixed with the intestinal contents (IC) prepared from untreated rats and the mixture was incubated anaerobically under an atmosphere of nitrogen at 37 C. Samples of the reaction mixture were removed at intervals to assay their mutagenic potential, to determine the radioactivity bound to the IC, and for analysis of the biliary metabolites. The binding of the radioactivity to the IC increased linearly as a function of time during the 1‐hr incubation. The time‐dependent binding does not occur with heat‐treated IC and the binding was inhibited by addition of D‐saccharic acid 1,4‐lacton, a β‐glucuronidase inhibitor. The mutagenicity (for Salmonella typhimurium strain TA98 without S9 mix) of the bile increased early in the incubation period and then decreased very rapidly. The mutagenicity of the bile was also enhanced by treatment with a sonicated IC extract or β‐glucuronidase, but not with a heat‐treated IC or arylsulfatase. The metabolites produced after the bile was incubated for short periods with the IC were mainly nitrohydroxypyrenes; at later times nitroreduction occurred. The level of acetylaminohydroxypyrenes, which were formed by deconjugation, did not change during the incubation. To determine the degree of contribution of the IC to the total acetylating capacity, we measured acetyltransferase activity of the IC and various organs in Wistar rats. The liver had the highest N‐acetyltransferase activity among the seventeen organs examined. Considerable activity was also detected in the kidney, small intestine, lung, and testis, but the IC showed very low activity. The acetylating capacity of the IC was 0.27% of the total capacity in rats, and that of the liver was more than 80%. These results suggest that the nitrohydroxypyrenes formed from 1‐NP in the liver were conjugated to glucuronic acid and excreted via the bile duct into intestine. Hydrolysis of these glucuronide conjugates by bacterial β‐glucuronidase liberated into intestine, free nitrohydroxypyrenes, which were direct‐acting mutagens. The released aglycons were then rapidly nitro‐reduced by intestinal microflora, but contribution of the intestinal microflora to acetylation of the reduced metabolites is very low.
Follow‐Up Study of Type C Influenza Outbreak in a Children's Home
Authors: Katagiri Susumu , Ohizumi Akiko , Ohyama Shinobu , Homma Morio
Published: November 2013

A follow‐up study of type C influenza in a children's home was made where the first outbreak of type C influenza occurred in March, 1981. During the survey period of 2 years, 2 additional outbreaks occurred in April and October, 1982 and 4 cases of the secondary and 2 cases of the tertiary infections were serologically confirmed. All of the children exposed to the outbreaks, except 2 particular cases who were suffering from German measles, showed similar mild respiratory symptoms characterized by fever and long‐lasting nasal discharge, irrespective of primary, secondary, and tertiary infections. No case of inapparent infection was observed. Incubation period was estimated to be not longer than 5 days and the period of virus shedding, to be longer than 22 days. Antibody response in the sera of patients to type C influenza virus was equally good after every infection, although it was not efficient to prevent the succeeding infection. Factors involved in the repeated infections of type C influenza were discussed.
Purification and Characteristics of Glucocorticoid Antagonizing Factor in Endotoxemia
Authors: Sakaguchi Shuhei , Yokota Katsushi
Published: November 2013

The present study involved the purification of GAF (glucocorticoid antagonizing factor) released in blood of endotoxemic mice, using the inhibition rate of tryptophan oxygenase (TO) activity in the mice liver as a parameter, to determine if this plays a role in metabolic disorders. GAF‐rich serum in zymosan‐primed and endotoxin‐injected mice was subjected to chromatography on DEAE‐Sepharose CL‐6B, Blue Sepharose CL‐6B and Sephadex G‐200 superfine columns. Finally, GAF fractions were purified by chromatography on a DEAE‐Sepharose CL‐6B column. The purified GAF showed a single band in electrophoresis in sodium dodecyl sulfate (SDS) polyacrylamide gel. The molecular weight of GAF was estimated to be 90,000. The purified GAF was regarded as glycoprotein. No factor (100 μg) exhibited lethal action on mice. The activity of TO in cortisone treated mice after injection of purified GAF was markedly lower than that in cortisone alone treated mice. On the other hand, there were no differences in tyrosine aminotransferase activities between the GAF plus cortisone injected group and cortisone only treated group. The glucose level after injection of GAF in cortisone treated mice initially showed hyperglycemia, but declined toward hypoglycemia 2 hr after injection, and thereafter returned nearly to the normal range by 4 hr. The liver glycogen level in GAF plus cortisone‐treated mice was markedly lower than that in cortisone‐alone treated mice.
Induction of Resistance in Mice by the Capsular Polysaccharide Antigens of Staphylococcus aureus
Authors: Yoshida Kosaku , Umeda Akiko , Ohshima Yukio
Published: November 2013

Mice actively immunized with capsular polysaccharides extracted from capsular type strains A, B, C, and D, determined by the serum‐soft agar technique, were protected against lethal infection by homologous strains, but no animals survived infection by heterologous substance immunization even with at high doses. Passive protective antibody in rabbit antisera prepared using these strains was absorbed out only by homologous capsular polysaccharide in mice. These results indicated that resistance was specific for capsular polysaccharide. The substance contained mainly neutral sugar, small amounts of hexosamine, methyl‐pentose, and phosphate although these amounts varied depending on the capsular types strains.
Effects of FLONLIZER®, Ultraviolet Sterilizer, on Legionella Species Inhabiting Cooling Tower Water
Authors: Yamamoto Hiroyuki , Urakami Itsuo , Nakano Koji , Ikedo Masanari , Yabuuchi Eiko
Published: November 2013

Legionella pneumophila in sterile distilled water was not detected after ultraviolet irradiation by FLONLIZER®, a new‐type sterilizer, at a flow rate of 82.5 to 364.8 liters/hr. When irradiated by FLONLIZER® at a flow rate of under 324.0 liters/hr, no viable cells of legionellae, other heterotrophic bacteria and bacterivorous protozoa were detected in the cooling tower water, which was found to contain L. pneumophila. No viable cells of L. pneumophila and L. bozemanii suspended in sterile distilled water were detected after the irradiation with UV‐doses of over 6.16 × 103 μW·sec/cm2. At the irradiation of low UV‐doses under 1.06 × 104 μW·sec/cm2, the viable count of legionellae recuperated by photoreactivation from UV‐damage increased with the exposure time under a white fluorescent lamp. However, in the samples irradiated with UV‐doses of over 3.52 × 104 μW·sec/cm2, equal to the FLONLIZER®, legionellae did not recuperate even after 18 hr illumination with a white fluorescent lamp. FLONLIZER® is thus expected to act as a sterilizer which can control the legionellae inhabiting cooling tower systems placed in outdoor space.
Characterization of Murine Monoclonal Antibodies to Human Interferon‐Gamma (IFN‐γ) and Their Application for Sandwich Enzyme‐Linked Immunosorbent Assay (ELISA)
Authors: Jitsukawa Tomofumi , Nakajima Satoko , Sugawara Isamu , Mori Shigeo , De Ley Marc
Published: November 2013

The three murine monoclonal antibodies (MAb), D1G2, D9D10, and D13C8, are specific for human interferon‐γ (IFN‐γ), but not human IFN‐α and IFN‐β. They react weakly with heat‐treated IFN‐γ. The three antibodies recognize different epitopes of the IFN‐γ molecule, as evaluated by antibody‐binding inhibition experiments. We have used these three monoclonal antibodies to construct a sandwich enzyme‐linked immunosorbent assay (ELISA). The best result was obtained when we used D1G2 or D9D10 MAb as a solid‐phase immunosorbent and D1G2 or D9D10 MAb as a tracer. When we measured IFN‐γ in sera by a combination of D1G2 (a solid‐phase) and D1G2 (a tracer), a result similar to the one by a combination of D9D10 (a solid‐phase) and D1G2 (a tracer), was obtained. This may suggest that human IFN‐γ exists in oligomeric form. Recombinant human IFN‐γ expressed in E. coli is detectable at a concentration of 1 ng/ml in this sandwich ELISA. This assay can be employed for the analysis of the structural characteristics of the human IFN‐γ molecule as well as measurement of IFN‐γ in human sera and tissue culture fluids.
Enzymic Detection of Adhesion of Enteropathogenic Escherichia coli to HEp‐2 Cells
Authors: Minami Junzaburo , Okabe Akinobu , Hayashi Hideo
Published: November 2013

We established a new method for detecting enteropathogenic Escherichia coli adhering to HEp‐2 cells. An essential part of the method is an assay of β‐galactosidase activity of adhered bacterial cells. It consisted of the following steps: (1) culture of bacterial cells in a medium containing isopropyl‐thio‐β‐d‐galactoside, an inducer of β‐galactosidase; (2) incubation of a bacterial culture with monolayered HEp‐2 cells in a 96‐well culture plate; (3) washing wells to remove bacterial cells which did not adhere to HEp‐2 cells, and (4) enzymic reaction for β‐galactosidase assay in wells. The strains used differed widely in their β‐galactosidase activities. However, a calibration curve for the enzyme activity, obtained from each bacterial sample, showed that 105 bacteria per well permitted an accurate estimation. The enzyme activity of adhered bacteria to the monolayered cells showed that 107 bacteria were appropriate for the adherence assay. The number of adhered bacteria thus obtained was in good agreement with a viable cell count. The result indicates that the new method is more reliable than a widely used method, counting the number of bacteria under a microscope. The present method also makes it easy to detect adherent strains of E. coli in large numbers of specimens.
Purification of a k‐Carrageenase from Marine Cytophaga Species
Authors: Sarwar Golam , Matayoshi Seiken , Oda Hiroshi
Published: November 2013

A mixture of extracellular carrageenases was isolated from the cell‐free medium of a culture of marine Cytophaga sp. 1k‐C783 grown on ZoBell 2216 E broth with 0.1% commercial carrageenan. A single active peak of k‐carrageenase was separated and purified from the mixture by ammonium sulfate precipitation, ion‐exchange chromatography, and Sephadex G‐200 gel filtration chromatography. Molecular weight of the purified k‐carrageenase was estimated as 100,000 by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). The purified k‐carrageenase had pH optimum 7.6 and temperature optimum 25 C.
Inhibitory Activity of (E)‐5‐(2‐Bromovinyl)‐2′‐Deoxyuridine on the Salmonid Herpesviruses, Oncorhynchus masou Virus (OMV) and Herpesvirus salmonis
Authors: Kimura Takahisa , Nishizawa Toyohiko , Yoshimizu Mamoru , De Clercq Erik
Published: November 2013

The highly potent and selective anti‐herpesvirus agent, (E)‐5‐(2‐bromovinyl)‐2′deoxyuridine (BVdU), was examined for its inhibitory effect on the salmonid herpesviruses Oncorhynchus masou virus (OMV) and Herpesvirus salmonis (H. salmonis). Minimum inhibitory concentrations (MIC) of BVdU for OMV and H. salmonis were 1.25 and 3.0 μg/ml, respectively; these values were equal to or higher than those obtained for acyclovir or cytarabin. OMV DNA polymerase activity was reduced in a dose‐dependent fashion by BVdU 5′‐triphosphate (BVdUTP) within the concentration range of 3 to 30 μm. However, BVdUTP could also be substituted for the natural substrate, TTP, in the OMV DNA polymerase assay. It is postulated that the inhibitory action of BVdU on the salmonid herpesviruses is more or less similar to that on other herpesviruses and resides with respect to the inhibition of the virus DNA polymerase activity as well as incorporation of BVdU into the viral DNA.
Screening of Bacteria with Antiviral Activity from Fresh Water Salmonid Hatcheries
Authors: Kamei Yuto , Yoshimizu Mamoru , Ezura Yoshio , Kimura Takahisa
Published: November 2013

Bacteria isolated from two salmonid hatcheries were screened for antiviral activity against infectious hematopoietic necrosis virus (IHNV) to ascertain the presence of bacteria with anti‐IHNV activity in the aquatic environment. Out of 710 bacterial isolates from the water and sediment samples, 190 strains showed anti‐IHNV activities of more than 50% plaque reduction. These antiviral activities were detected predominantly in Pseudomonas, Aeromonas/Vibrio, and coryneforms. In one hatchery, the bacteria with antiviral activities were more prevalent in sediment samples than in water samples. Seventy‐seven percent of the isolates with higher antiviral activities (>90% plaque reduction) belonged to Pseudomonas.
Chemotaxis for Methamphetamine in Escherichia coli
Authors: Fukunaga Shinya , Horiguchi Hiromi , Iwamoto Masayuki , Shigenobu Youji , Shinozawa Takao
Published: November 2013

Escherichia coli showed the chemotactic response for methamphetamine, a biogenic stimulative amine. The response was qualitatively detected by using a paper disk filter. The quantitative assay showed that E. coli responded to methamphetamine at 10−5 m or more.
Variation of Virulence and Other Properties among Sendai Virus Strains
Authors: Yamaguchi Ryoji , Iwai Hiroshi , Ueda Katsumoto
Published: November 2013

The virulence of five Sendai virus strains (MN, Z, KN, Mol, and Hm) isolated from laboratory rodents was compared, using 3‐week‐old female Jcl‐ICR mice. The virulence of the strains was Mol, MN, KN, Z, and Hm in decreasing order. The 50% lethal dose and 50% lung consolidation inducing dose of the highest virulent strain differed by the order of more than 103 and 106, respectively, from those of the lowest virulent one. Other properties such as the growth rate in LLC‐MK2 cells, neuraminidase activities, and molecular weights of structural proteins also differed among the virus strains. These results indicate that Sendai virus prevailing in laboratory rodents is not homogenous with respect to virulence and some other properties.
Chemotactic Response of Human Neutrophils to N‐Acetyl Chitohexaose in vitro
Authors: Tokoro Akio , Suzuki Ko , Matsumoto Tatsuji , Mikami Takeshi , Suzuki Shigeo , Suzuki Masuko
Published: November 2013

N‐Acetyl chitohexaose (NACOS‐6) was able to display chemotactic response of human neutrophils in vitro. In order to analyze the mechanism, a series of chemotaxis studies by means of neutrophils treated with inhibitors of phospholipase A2, cyclooxygenase, or lipoxygenase to NACOS‐6 was conducted. The treatment of neutrophils with inhibitors of phospholipase A2 or cyclooxygenase resulted in decrease of number of migrated cells. However, the lipoxygenase inhibitors did not exhibit the same effect. On the other hand, the treatment of neutrophils with inhibitors of phospholipase A2 or lipoxygenase resulted in decrease of chemotactic response to Formyl‐Met‐Leu‐Phe (FMLP), although the cyclooxygenase inhibitors did not inhibit chemotaxis of neutrophils. Neutrophils added to exogenous prostaglandin E2 (PGE2) caused an enhanced chemotactic response to NACOS‐6. These results indicate that the mechanism of chemotactic response to NACOS‐6 was different from that of FMLP, and that the response was enhanced by PGE2 released from the neutrophils with stimulation of NACOS‐6.
Resistance of Pseudomonas pseudomallei to Normal Human Serum Bactericidal Action
Authors: Ismail Ghazally , Razak Nyonya , Mohamed Rahmah , Embi Noor , Omar Othman
Published: November 2013

The effect of human normal serum (HNS) on Pseudomonas pseudomallei was determined. It is apparent from our data that the organism is resistant to the normal serum bactericidal mechanism. Ancillary experiments to confirm this serum‐resistant property of P. pseudomallei were done by examining the effects of growth phase conditions of the bacteria (i.e., logarithmic and stationary phases) and different buffered systems used as diluent in our bactericidal assay. Results obtained showed similar degree of resistance to serum bactericidal killing by 5 strains of the organisms tested. The possible survival advantage of serum‐resistant property to P. pseudomallei as bacterial pathogens known to invade the blood stream is discussed.
Maternal Transmission and Dental Caries Induction in Sprague‐Dawley Rats Infected with Streptococcus mutans
Authors: Ooshima Takashi , Sumi Norio , Izumitani Akira , Sobue Shizuo
Published: November 2013

Thirty‐four female rats (18 days old) were infected with Streptococcus mutans MT8148R (serotype c) or 6715 (g). Diets containing different proportions of sucrose were used to prepare the dams which harbored various levels of S. mutans in their oral cavity. Around 66 days of age, the female rats were bred and 34 dams subsequently bore 322 offspring. The dams were killed upon weaning (20 days of age) of their respective litters. There were positive correlations between the recovery of inoculated S. mutans and the caries incidence in the dams. Transmission of S. mutans from a dam to her offspring was studied in 10‐, 15‐, 20‐, 27‐, 34‐, 41‐, 48‐, and 55‐day‐old rats by evaluating the recover of S. mutans from the offspring. Positive correlation between the magnitudes of recovered S. mutans MT8148R from dams and their offspring was found in all ages of young rats examined. Furthermore, caries incidence in young rats was found to be positively correlated with the recovery of both strains of S. mutans as well as with incidence of caries in their respective dams.
Production of Interleukin 1 Inhibitors by the Murine Macrophage Cell Line P388D1 Which Produces Interleukin 1
Authors: Isono Noriko , Kumagai Katsuo
Published: November 2013

A murine macrophage cell line P388D1 in in vitro culture without any specific stimulation produced both interleukin 1 (IL1) and IL1 inhibitor which inhibits mitogenic response of murine thymocytes to IL1 in the culture fluids. The factor(s) responsible for inhibiting IL1‐induced thymocyte proliferation consisted of at least two molecules: factor I (FI) with an isoelectric point of 6.0 and factor II (FII) with an isoelectric point of 5.3, both of which had a similar m.w. of 40–60 kDa. FI activity was sensitive to heat (56 C) treatment and acid pH (3.0) treatment, while FII was resistant to both treatments. Both FI and FII inhibited mitogenic responses of thymocytes to IL1, but not proliferation of murine lymphoid cells induced by other interleukins, namely, IL2, IL3, or IL4. Neither showed any inhibition of spontaneous proliferation of murine tumor cell lines, suggesting that inhibition was specific for IL1, but not nonspecifically inhibiting for cellular DNA. These IL1 inhibitors were also suggested to be acting in the early phase of interaction between IL1 and lymphoid cells. The possible role of these inhibitors as representatives of regulatory substances, which normally control IL1 activities either in the levels of inflammation or immune responses, was discussed.
Effect of Rice Fiber on Human Fecal Microflora
Authors: Benno Yoshimi , Endo Kimiko , Miyoshi Hiroko , Okuda Toyoko , Koishi Hideo , Mitsuoka Tomotari
Published: November 2013

The effects of brown rice, containing fourfold as much dietary fiber as polished rice, on the human fecal microflora were determined. Significantly increased numbers of Bifidobacterium adolescentis and Enterococcus faecalis were observed during the brown rice intake, whereas the total counts and the numbers of bacteroides, Eubacterium aerofaciens, and Escherichia coli during the intake were lower than those before and after the intake. Lower numbers of clostridia and low incidences of Clostridium paraputrificum and C. perfringens were observed during the brown rice intake.
Susceptibility of Mycobacterium avium and Mycobacterium intracellulare to Various Antibacterial Drugs
Authors: Tomioka Haruaki , Sato Katsumasa , Saito Hajime , Yamada Yoshitaka
Published: November 2013

Mycobacterium avium and M. intracellulare of human and natural sources, identified by the Gen‐Probe ®Rapid Diagnostic System for M. avium Complex (MAC) were studied for susceptibility to eight different drugs. In the case of human isolates of MAC, the following was noted. M. avium showed nearly the same susceptibility to streptomycin, kanamycin, ethambutol, and clofazimine as was seen with M. intracellulare. M. avium was much more resistant to rifampicin and rifabutin than was M. intracellulare, and M. avium was more susceptible to quinolones such as ofloxacin and ciprofloxacin. Conversely, in the case of MAC from natural sources, there was no difference between the susceptibility of M. avium and M. intracellulare to these antibacterial agents.
A Case of Dengue Hemorrhagic Fever in a Japanese Child
Authors: Okuno Yoshinobu , Harada Tokuzo , Ogawa Michiko , Okamoto Yutaka , Maeda Kazuhiro
Published: November 2013

A 6‐year‐old Japanese girl contracted a febrile illness with hemorrhagic manifestations when she traveled in Indonesia. A remarkable decrease in the numbers of platelets and white blood cells was observed in her acute‐phase blood specimens. Her father, who accompanied her, showed dengue fever‐like symptoms at almost the same time as her illness. It was determined by serological tests that they were infected with dengue virus type 1. Moreover, she showed a secondary antibody response to the flavivirus due to the pre‐existing antibody to Japanese encephalitis virus. This is the first confirmed case of dengue hemorrhagic fever (DHF) in Japanese people.
Purification of Cross‐Reacting Protein Antigen Shared by Yersinia enterocolitica and Other Gram‐Negative Bacteria with Monoclonal Antibody
Authors: Yamaguchi Hiroyuki , Taguchi Haruhiko , Katura Takuya , Kumada Junko , Uekusa Takeyuki , Ogata Sachio
Published: November 2013

A monoclonal antibody against the Yersinia enterocolitica 60‐kilodalton (kDa) antigen, designated cross‐reacting protein antigen (CRPA), was obtained by cell fusion. The CRPA common to gram‐negative bacteria was purified from Y. enterocolitica by the affinity chromatography with the monoclonal antibody (IgGl) thus obtained. The purified CRPA showed a single band of 60 kDa in SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE), and reacted with rabbit antisera against Y. enterocolitica, Vibrio cholerae, Escherichia coli, Pseudomonas aeruginosa, and Shigella sonnei in Western blot analysis. The monoclonal antibody, however, reacted with a 60 kDa peptide from Y. enterocolitica, but not with the antigens from other gram‐negative bacteria such as V. cholerae, E. coli, S. sonnei, Salmonella enteritidis, Serratia marcescens, Klebsiella pneumoniae, Proteus mirabilis, and P. aeruginosa. The results suggested that both species‐specific and cross‐reactive epitopes were present on a CRPA molecule.