bluuebunny/crossref_metadata_2025_split huggingface dataset:
Demonstration of Antigenic and Genotypic Variation in Orientia tsutsugamushi Which Were Isolated in Japan, and Their Classification into Type and Subtype
Authors: Ohashi Norio , Koyama Yoko , Urakami Hiroshi , Fukuhara Masahiro , Tamura Akira , Kawamori Fumihiko , Yamamoto Seigo , Kasuya Shiro , Yoshimura Kentaro 
Published: November 2013

AbstractA total of 40 strains of Orientia tsutsugamushi (34 isolates from patients and trombiculid mites in Japan, and 6 prototype strains of antigenic variants) were examined for classification based on the reactivities with type‐specific monoclonal antibodies in indirect immunofluorescence tests, and on the restriction fragment length polymorphism of a polymerase chain reaction (PCR)‐amplified 56‐kilodalton type‐specific antigenic protein gene. By these methods, several antigenic and genotypic variants were found among the strains, and these variants were classified into types and further into subtypes. These results suggest that there are many variants in O. tsutsugamushi, and the methods used here seem to be useful for the systematic classification of the numerous variants. A strain which may be a new type distinguishable from those identified previously was also found in this study. Furthermore, variety in the degree of pathogenicity in mice related to type and/or subtype classification were observed.
Characterization of Actinobacillus actinomycetemcomitans Hemolysin
Authors: Kimizuka Ryuta , Miura Tadashi , Okuda Katsuji 
Published: November 2013

AbstractTwenty out of 33 Actinobacillus actinomycetemcomitans strains formed hemolytic colonies on horse blood agar plates under anaerobic conditions. The hemolytic activity found in A. actinomycetemcomitans strain 137HE was examined. This activity was detected in the late exponential to early stationary phases of growth. Human erythrocytes were the most susceptible, followed by rabbit, sheep, horse and swine red blood cells. The majority of activity was detected in the cell‐associated vesicle fraction. Zwitterionic detergent 3‐[(3‐cholamidopropyl)‐dimethyl‐ammonio]‐1‐propanesulfonate (CHAPS) extract from whole cells was semipurified by ammonium sulfate precipitation, preparative isoelectric focusing (IEF) and gel‐filtration chromatography to yield a major band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) with a molecular mass of 12 kDa. Heating at 80 C for 30 min and treatment with proteinase K or trypsin resulted in complete disappearance of the hemolytic activity. Sulphydryl reagents enhanced activity and small amounts of cholesterol inhibited it. In summary, we demonstrated the presence of hemolysin in A. actinomycetemcomitans, and examined and characterized it.
Electron Microscopy of the Major Outer Membrane Protein of Campylobacter jejuni
Authors: Amako Kazunobu , Wai Sun Nyunt , Umeda Akiko , Shigematsu Mika , Takade Akemi 
Published: November 2013

AbstractThe surfaces of the disrupted‐cell surfaces of the Campylobacter jejuni strains FUM158432 and M1 were examined using the negative‐staining technique and electron microscopy. The surfaces of the whole cells and the outer membranes were covered with small dark dots which, in some areas, were arranged in hexagonal patterns. The hexagonal arrangement was more clearly seen in extracted outer membrane. The size of each structure was measured based on a center‐to‐center distance with the adjacent structure, and was determined to be 9.9±0.9 nm. A profile of the proteins in the outer membrane by SDS‐PAGE, performed in 0.1% SDS and at 100 C, showed 42 kDa proteins to comprise the major outer membrane protein of this bacterium. Digestion of the outer membrane materials with proteinase reduced this protein band in the SDS‐PAGE, and the amount of dark dots on the electron micrograph indicated the structure to be the major outer membrane protein (porin) of this bacterium. The power spectrogram of a computer‐assisted Fourier transformation of the hexagonally arranged porin proteins suggests that the porin has a trimeric structure rather than a monomeric one.
Seroepidemiology of Feline Chlamydiosis by Microimmunofluorescence Assay with Multiple Strains as Antigens
Authors: Pudjiatmoko  , Fukushi Hideto , Ochiai Yoshitsugu , Yamaguchi Tsuyoshi , Hirai Katsuya 
Published: November 2013

AbstractThe prevalence of anti‐chlamydia antibodies was examined in 232 cat sera collected in 1985 and from 1993 to 1995 from laboratories and veterinary hospitals located in 11 prefectures of Japan. The antibodies were determined by an indirect microimmunofluorescence test using six strains of feline Chlamydia: one strain each of avian‐ and guinea pig‐derived C. psittaci and one strain each of C. pecorum, C. pneumoniae and C. trachomatis. Positive rates of IgG antibodies to chlamydiae were 34.4% in 1985 and 16.5–21.4% from 1993 to 1995. Positive rates of IgM antibodies to chlamydiae were 8.2% in 1985 and 6.6–14.3% from 1993 to 1995. Variations in antibody reactivity to the different feline strains were observed. The results suggest the wide prevalence of chlamydial infection in cats in Japan, and antigenic diversity in the feline strains of C. psittaci.
Detection and Identification of Yersinia pestis by Polymerase Chain Reaction (PCR) Using Multiplex Primers
Authors: Tsukano Hiroko , Itoh Ken‐Ichiro , Suzuki Sosuke , Watanabe Haruo 
Published: November 2013

AbstractA PCR method for detection of Yersinia pestis‐virulence determinants by the use of multiplex primers was developed. Four pairs of oligonucleotide primers were designed from each gene of three kinds of virulent plasmids and a chromosomal DNA; 60‐Md plasmid‐located gene (caf1) encoding Y. pestis‐specific capsular antigen fraction 1, a Y. pestis‐specific region of a yopM gene encoded on 42‐Md virulent plasmid, a plasminogen activator gene (pla) encoded on Y. pestis‐specific 7‐Md plasmid and an invasin protein gene (inv) encoded on chromosomal DNA. This multiplex‐primer system was specific for the detection of Y. pestis among pathogenic Yersinia species and other enterobacteriaceae having antigens common to Y. pestis. Since this method is simple and safe, it will be useful to identify and confirm Y. pestis in cases of emergency and for the surveillance of epidemics.
Migration of Salmonella typhi through Intestinal Epithelial Monolayers: An In Vitro Study
Authors: Kops Sandra Kraeuter , Lowe Daniel K. , Bement William M. , West A. Brian 
Published: November 2013

AbstractThis study characterizes the transmigration of enteroinvasive Salmonella typhi in vitro, using a human intestinal epithelial cell line as a model of small intestinal epithelium. C2BBe cells, a subclone of CACO‐2 with a highly differentiated enterocytic phenotype, were grown to maturity on Transwell filters. S. typhi Ty2 and the vaccine strain, Ty21a, the S. typhi mutant X7344 and parent strain SB130, and S. typhimurium 5771 in logarithmic phase were introduced to the upper chamber of the filter units. Numbers of bacteria in the lower chamber, TER and permeability of the monolayer to mannitol were measured over time. Monolayers were examined by light, electron and confocal microscopy to determine the pathway of bacterial transmigration, and intracellular bacteria were estimated by gentamicin assay. Epithelial cell injury was quantified by light microscopy. S. typhi transmigrated earlier and in larger numbers than S. typhimurium, inducing marked changes in electrical resistance and permeability. Unlike S. typhimurium, S. typhi selected epithelial cells in small number and caused their death and extrusion from the monolayers leaving holes through which S. typhi transmigrated. Ty2 consistently transmigrated in larger numbers and with more injury to monolayers than Ty21a. S. typhi crosses the monolayers of C2BBe cells by a paracellular route in contrast to the transcellular pathway described for other Salmonellae. This may be related to the unique pathophysiology of S. typhi infection and the restricted host specificity of this pathogen. In these assays the vaccine strain, Ty21a, is slightly less invasive than its parent, though more invasive than S. typhimurium.
Cooperative Anti‐Candida Effects of Lactoferrin or Its Peptides in Combination with Azole Antifungal Agents
Authors: Wakabayashi Hiroyuki , Abe Shigeru , Okutomi Takafumi , Tansho Shigeru , Kawase Kouzou , Yamaguchi Hideyo 
Published: November 2013

AbstractThe effects of lactoferrin (LF), an antimicrobial protein secreted in body fluids, and its peptides in combination with azole antifungal agents were investigated by the micro‐broth‐dilution method in a study of Candida albicans. In the case of LF, its pepsin hydrolysate (LFhyd) or the LF‐derived antimicrobial peptide Lactoferricin® B (LF‐B), the concentrations required to inhibit the growth of Candida decreased in the presence of relatively low concentrations of clotrimazole (CTZ). The minimum inhibitory concentration (MIC) of all azole antifungal agents tested was reduced by 1/41/16 in the presence of a sub‐MIC level of each of these LF‐related substances. Polyene and fluoropyrimidine antifungal agents did not show such a combined effect with these LF‐related substances. The anti‐Candida activity of LF or LF‐B in combination with CTZ was shown to be synergistic by checkerboard analysis. These results indicate that LF‐related substances function cooperatively with azole antifungal agents against C. albicans.
Production, Binding Characteristics and Protective Immunity of Monoclonal Antibody to Pneumococcal Type‐9V Conjugate
Authors: Lee Chi‐Jen , Karpas Arthur , Wang Theresa R. , Kosaka Tadashi , Koizumi Kaio 
Published: November 2013

AbstractA monoclonal antibody (MAb) to pneumococcal type‐9V polysaccharide (PS) was produced using PS conjugated to inactivated pneumolysin as the immunogen. The MAb to 9V PS was of the IgG1 subclass. The antigen‐antibody reaction increased rapidly at low concentrations and reached a plateau at 10 μg PS/ml as measured by nephelometry of the group 9 PS against 9V MAb binding. In contrast, the binding of group 9 PS against rabbit 9V antiserum (AS) increased proportionally and continued to increase up to the highest concentration of PS tested (20 μg PS/ml). The 9V MAb reacted with all group 9 PSs (9A, 9L, 9N and 9V) by immunodiffusion. In the homologous 9V Ag‐MAb reaction, there were marked differences in the inhibition of binding by the cross‐reactive 9L PS (19.2% inhibition) and the 9N PS (0.2%). In contrast, inhibition of the homologous 9V Ag‐rabbit AS binding by cross‐reactive 9L and 9N PSs ranged from 57.8 to 62.7%. Removal of the O‐acetyl group from 9V PS by alkali hydrolysis resulted in decreased binding with rabbit 9V AS. However, the binding reaction with 9V MAb was less affected by the loss of O‐acetyl content. The 9V MAb was both opsonic and passively protected young mice against challenge with type‐9V pneumococci.
An Epidemiological Study of Salmonella enteritidis by Pulsed‐Field Gel Electrophoresis (PFGE): Several PFGE Patterns Observed in Isolates from a Food Poisoning Outbreak
Authors: Murase Toshiyuki , Nakamura Akiko , Matsushima Akiyoshi , Yamai Shiro 
Published: November 2013

AbstractAn epidemiological analysis of Salmonella enteritidis from a food poisoning was done using pulsed‐field gel electrophoresis (PFGE) of BlnI‐ or XbaI‐digested fragments of chromosomal DNA of isolates. S. enteritidis isolates obtained from 19 patients had identical PFGE patterns. Therefore, a strain giving the same pattern was considered to be the causative agent of this outbreak. In addition, four isolates that had different BlnI‐digested PFGE patterns were obtained from three patients, suggesting that the observed variations in PFGE patterns might occur as the result of some point mutations of chromosomal DNA during growth or from the existence of several S. enteritidis strains from various sources. Subsequent PFGE analysis of continuously subcultured strains supported the former possibility. These observations indicate that PFGE analysis on multiple numbers of colonies from each patient are necessary for the epidemiologic investigation of S. enteritidis.
Differential Expression of a Mycobacterium tuberculosis Protein Recognized by a Mouse Monoclonal Antibody
Authors: Cummings Patrick J. , Rowland Sharon S. , Hooper Nancy E. , Schwalbe Richard S. 
Published: November 2013

AbstractMurine monoclonal antibodies were produced against Mycobacterium tuberculosis (Mtb) using standard hybridoma procedures. By a whole cell enzyme‐linked immunosorbent assay (ELISA), one monoclonal antibody (mAb), HB28, demonstrated high level specific reactivity to Mtb. Western blot analysis demonstrated reactivity to a single 65 kDa Mtb protein in the cell wall extract and culture filtrate. HB28 mAb appears to be recognizing a 65 kDa Mtb protein that is over‐expressed by Mtb but not other species under certain culture conditions. Differential expression and detection of this protein by HB28 mAb may have potential for diagnostic applications.
Rapid Increase of Pneumococcal Resistance to β‐Lactam and Other Antibiotics in Isolates from the Respiratory Tract (Nagasaki, Japan: 1975–1994)
Authors: Rikitomi Naoto , Sow Papa Salif , Watanabe Kiwao , Nunez Duilio S. , Martinez Glenda , Nagatake Tsuyoshi 
Published: November 2013

AbstractThe susceptibility of 101 pneumococcal isolates from the respiratory tract during 1991–1994 was examined and compared with the susceptibility of isolates over the period of 1975–1990. A rapid increase of resistance was seen not only to penicillin but also other antimicrobial agents. During 1991–1994, 38% of all the isolates were resistant to penicillin. The rates of resistance during this period were 16–23% for three newer cephalosporins, 18% for imipenem, 69% for tetracycline, 31% for erythromycin, 20% for chloramphenicol and 9% for clindamycin. The use of antibiotics within one month prior to pneumococcal isolation was correlated with penicillin resistance (P < 0.05). Serotyping of the isolates by antiserum revealed differences in predominant types between penicillin‐resistant (19F, 23F, 4) and ‐susceptible isolates (15, 4, 11A). Our data suggests that anti‐pneumococcal antibiotics should be carefully chosen on the basis of susceptibility tests.
Expression of thecolHGene EncodingClostridium histolyticumCollagenase inBacillus subtilisand Its Application to Enzyme Purification
Authors: Jung Chang‐Min , Matsushita Osamu , Katayama Seiichi , Minami Junzaburo , Ohhira Lichiro , Okabe Akinobu 
Published: November 2013

AbstractThecolHgene encoding 116‐kDa collagenase ofClostridium histolyticum(cColH) was cloned into anEscherichia coli‐Bacillus subtilisshuttle vector to develop a method for purification of recombinant collagenase (rColH). When plasmid pJCM310 containing thecolHgene was introduced intoB. subtilisDB104 and the transformant was grown in LB broth at 37 C, stability of the plasmid was not maintained. However, stability was partly improved by growing the transformant in a modified LB broth containing 0.5Msodium succinate with gentle shaking at 35 C. When the transformant was grown to an optical density of 0.4 at 600 nm in this medium, pJCM310 was stable and rColH was produced in sufficient amounts. rColH was purified to homogeneity by ammonium sulfate precipitation, gel filtration and ion‐exchange chromatography. The yield of rColH from an 800‐ml culture was 0.53 mg and its specific activity was estimated to be 1,210 U per mg of protein. The purified rColH was capable of degrading native type‐I collagen fibril from bovine achilles tendon, as was demonstrated by zymography. A comparison of the N‐terminal amino acid sequence between cColH and rColH revealed that rColH has 10 extra N‐terminal amino acid residues. However, the peptide mapping of rColH with V8 protease was virtually identical to that of cColH. Furthermore, the molecular mass of rColH was estimated to be 112,999 Da by mass spectrometry, coinciding with the value of 112,977 Da, which was predicted from the nucleotide sequence of thecolHgene. Therefore, the recombinantB. subtilisculture is capable of serving as a useful source for enzyme purification.
Effects of Granulocyte‐Macrophage Colony Stimulating Factor (GM‐CSF) on Biomaterial‐Associated Staphylococcal Infection in Mice
Authors: Rózalska Barbara , Ljungh Asa , Paziak‐Domańska Beata , Rudnicka Wieslawa 
Published: November 2013

AbstractStaphylococcal infections are a major complication in the usage of biomaterials. Different modifications of polymers have been made to reduce the incidence of such infections. We studied the effects of modifying heparinized polyethylene (H‐PE) with mouse recombinant granulocyte‐macrophage stimulating factor (rGM‐CSF). The elimination of staphylococci (Staphylococcus aureus, S. epidermidis) from the peritoneum of mice implanted with rGM‐CSF‐coated H‐PE was slightly more effective than the elimination of the bacteria from the peritoneum of animals implanted with uncoated H‐PE. Most interestingly, the number of staphylococci present in the biofilms covering rGM‐CSF‐coated implants were significantly lower than the number of bacteria detected on the surface of H‐PE not coated with rGM‐CSF. In vitro, rGM‐CSF restored the anti‐bacterial potency of the phagocytes, which had been reduced by surface contact with H‐PE. The results suggest that modification of biomaterials with rGM‐CSF could be one way of preventing staphylococcal infections; especially in neutropenic disorders, which constitute the highest risk factor for foreign body‐associated infections.
The Thermostable Direct Hemolysin Gene (tdh) of Vibrio hollisae Is Dissimilar in Prevalence to and Phylogenetically Distant from the tdh Genes of Other Vibrios: Implications in the Horizontal Transfer of the tdh Gene
Authors: Nishibuchi Mitsuaki , Janda J. Michael , Ezaki Takayuki 
Published: November 2013

AbstractVibrio hollisae strains isolated recently from patients in various locations were examined for the presence of the thermostable direct hemolysin gene (tdh) using nucleic acid hybridization and polymerase chain reaction assays. The results were consistent with the previous finding that all strains of V. hollisae carry the tdh gene. In contrast, the tdh gene has been detected in a minority of strains for other Vibrio species (V. parahaemolyticus, V. cholerae non‐O1, and V. mimicus). Detailed phylogenetic analysis showed that the tdh genes of the non‐V. hollisae species were very closely related to each other and that the tdh gene of V. hollisae was distantly related to the tdh genes of the non‐V. hollisae species. These results and the proposed insertion sequence‐mediated tdh transfer mechanism suggest that the tdh gene may have been maintained stably in V. hollisae and that the tdh genes of the non‐V. hollisae species may have been involved in recent horizontal transfer.
Cloning and Sequencing of the Spore Germination Gene of Bacillus megaterium ATCC 12872: Similarities to the NaH‐Antiporter Gene of Enterococcus hirae
Authors: Tani Katsuji , Watanabe Takuya , Matsuda Hideyasu , Nasu Masao , Kondo Masaomi 
Published: November 2013

AbstractThe germination mutant TM‐31 of Bacillus megaterium ATCC 12872, was isolated by transposon Tn917 insertional mutagenesis. Glucose, L‐proline, L‐leucine and KNO3 germinated TM‐31 poorly. The DNA in the region of the Tn917 insertion was cloned, and its nucleotide sequence determined. One major open reading frame was present on the cloned DNA. The hydrophobic protein encoded is presumably membrane‐associated. A homology search revealed that the gene encoded in the region of the Tn917 insertion is homologous to napA of Enterococcus hirae. napA codes for the NaH‐antiporter. It is hypothesized that transport of cations must play an important role in spore germination in B. megaterium ATCC 12872.
Prophylactic Effect of Enterococcus faecalis FK‐23 Preparation on Experimental Candidiasis in Mice
Authors: Satonaka Katsuhito , Ohashi Kazutomo , Nohmi Takao , Yamamoto Tetsuro , Abe Shigeru , Uchida Katsuhisa , Yamaguchi Hideyo 
Published: November 2013

AbstractThe prophylactic effects of heat‐killed cells of Enterococcus faecalis FK‐23 (FK‐23 preparation) on experimental candidiasis were investigated in normal and leukopenic mice. In cyclophosphamide‐induced leukopenic mice, oral or intraperitoneal administration of the FK‐23 preparation at a daily dose of 1.25 or 5 mg/mouse for 3 consecutive days prior to Candida albicans infection significantly prolonged survival periods of the infected mice, and decreased viable counts of C. albicans recovered from their kidneys. In normal mice, the FK‐23 preparation administered at dosages ranging from 0.63 to 10 mg/mouse/day for 3 consecutive days was ineffective, while in leukopenic mice, the FK‐23 administered orally caused a facilitated recovery in the number of white blood cells including neutrophils. Furthermore, intraperitoneal administration of the FK‐23 preparation into mice augmented the anti‐Candida activity of immunocompromised peritoneal exudate cells obtained from the animals. These results suggested the potential usefulness of the FK‐23 preparation as a prophylactic agent for the management of patients with opportunistic fungal infections.
Susceptibility of Helicobacter pylori Isolates against Agents Commonly Administered for Eradication Therapy and the Efficacy of Chemotherapy
Authors: Gotoh Akira , Kawakami Yoshiyuki , Akahane Takayuki , Akamatsu Taiji , Shimizu Toshiki , Kiyosawa Kendo , Katsuyama Tsutomu 
Published: November 2013

AbstractAlthough chemotherapy has been documented to be effective in the treatment of Helicobacter pylori‐associated gastritis and gastroduodenal ulcers, some cases are known to have been unsuccessful in the attempt to eradicate this species. In this study, we examined the relation between the susceptibility of H. pylori isolates and the efficacy of chemotherapy. We utilized the modified agar plate dilution method to determine the minimum inhibitory concentrations (MICs) of 63 H. pylori strains isolated before treatment with several drugs routinely used during eradication chemotherapy. Among the drugs tested, amoxicillin (AMPC) and clarithromycin (CAM) demonstrated high degrees of activity with MIC90, 0.39 and 0.2 μg/ml, respectively. No highly resistant strain against AMPC was detected among the strains examined, while for CAM and metronidazole (MTZ), 9.5% and 7.9% of the strains, respectively, were resistant before treatment. It should be noted that all of the MICs of the strains from patients with successful therapy were lower than those from patients with unsuccessful therapy. These findings indicate that susceptibility tests should be carried out prior to the commencement of drug administration in order to provide safer and more effective chemotherapy.