- Volume 64, Issue 10, 2015
Volume 64, Issue 10, 2015
- Review
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Current concepts in the pathogenesis and treatment of chronic suppurative otitis media
Otitis media (OM) is an inflammation of the middle ear associated with infection. Despite appropriate therapy, acute OM (AOM) can progress to chronic suppurative OM (CSOM) associated with ear drum perforation and purulent discharge. The effusion prevents the middle ear ossicles from properly relaying sound vibrations from the ear drum to the oval window of the inner ear, causing conductive hearing loss. In addition, the inflammatory mediators generated during CSOM can penetrate into the inner ear through the round window. This can cause the loss of hair cells in the cochlea, leading to sensorineural hearing loss. Pseudomonas aeruginosa and Staphylococcus aureus are the most predominant pathogens that cause CSOM. Although the pathogenesis of AOM is well studied, very limited research is available in relation to CSOM. With the emergence of antibiotic resistance as well as the ototoxicity of antibiotics and the potential risks of surgery, there is an urgent need to develop effective therapeutic strategies against CSOM. This warrants understanding the role of host immunity in CSOM and how the bacteria evade these potent immune responses. Understanding the molecular mechanisms leading to CSOM will help in designing novel treatment modalities against the disease and hence preventing the hearing loss.
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- Pathogenicity and Virulence/Host Response
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Patterns of EPIYA motifs among cagA-positive Helicobacter pylori strains: a case–control study in a Turkish population with Eurasian geographical features
Geographical variation in the frequency of various gastroduodenal pathologies was shown to be related to the geographical diversity of H. pylori CagA Glu-Pro-Ile-Tyr-Ala (EPIYA) patterns. We examined the EPIYA patterns of H. pylori and the association of EPIYA patterns with gastric cancer (GC) for the first time, to the best of our knowledge, in Turkey. The patient group (PG) contained 60 patients [38 GC and 22 duodenal ulcer (DU) patients]. The control group (CG) was 110 individuals [94 gastritis patients and 16 persons with a normal gastrointestinal system (NGIS)]. Specific primers were used for the detection of cagA including empty-site-positive and EPIYA-A, -B, -C, -D PCR. Bands of EPIYA-A, -B, -C were confirmed by DNA sequencing. One hundred and forty-two (83.5 %) strains [60 in the PG (38 GC, 22 DU), 82 in the CG (72 gastritis, 10 NGIS)] were positive for the cagA gene. EPIYA-C with multiple repeats was detected in 34 (23.9 %) strains, and 22 (64.7 %) were from GC patients. EPIYA-C with one repeat was detected in 89 (62.7 %) strains, and 54 (60.7 %) were from gastritis patients. EPIYT was detected in 10 strains, and EPIYA-D was not detected. The number of EPIYA-C with multiple repeats was significantly higher for the PG than for the CG (P < 0.0001). In GC patients, the number of EPIYA-C with multiple repeats was significantly higher than one repeat (P < 0.0001). In conclusion, our study showed that multiple EPIYA-C repeats increases the GC risk by 30.6-fold and the DU risk by 8.9-fold versus the CG. This indicates that Western-type H. pylori strains in Turkey have similar EPIYA motifs to those of neighbouring countries and Western populations.
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A new variant of cytolethal distending toxin in a clinical isolate of Campylobacter hyointestinalis
Increasing numbers of Campylobacter hyointestinalis have been isolated from humans and animals with gastroenteritis, although the virulence mechanism of this species remains largely unknown. Here, we show that C. hyointestinalis isolated from a patient with diarrhoea in Thailand produced a novel variant of cytolethal distending toxin (CDT). Sequencing of a 13 965 bp genomic region of C. hyointestinalis carrying the genes coding for Ch-CDT revealed three ORFs of 798, 804 and 537 bp, which code for the Ch-CdtA, Ch-CdtB and Ch-CdtC subunits, respectively. The deduced amino acid sequence of Ch-CdtA showed ∼38.9 % homology with the CdtA of Campylobacter coli, but sequences of Ch-CdtB and Ch-CdtC were homologous to CdtB (65.7 %) and CdtC (33.1 %) of Campylobacter upsaliensis, respectively. Filter-sterilized sonic lysate of C. hyointestinalis demonstrated distension and death of HeLa cells by arresting the cell cycle at the G2/M phase and phosphorylation of host histone H2AX, a sensitive marker of DNA double-strand breaks. Rabbit antiserum raised against recombinant Ch-CdtB was not reactive against the recombinant CdtB protein of Campylobacter jejuni. A reconstituted Ch-CDT holotoxin prepared using each of the recombinant subunit proteins demonstrated distension and death of HeLa cells, suggesting that the C. hyointestinalis isolate indeed produced functionally active Ch-CDT. Furthermore, the immunological distinctiveness of the Ch-CDT produced by C. hyointestinalis and the increasing prevalence of the species in patients and animals with gastroenteritis suggest that this species may be an important emerging zoonotic pathogen.
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- Diagnostics, Typing and Identification
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Characterization of the medically important yeast Trichosporon mucoides and its close sister Trichosporon dermatis by traditional and advanced technologies
More LessTrichosporon dermatis is a causative agent of several mycoses in immunocompromised patients but is often misidentified as Trichosporon mucoides due to their phenotypic resemblance. In order to evaluate the current identification keys for these species and to develop a rapid and reliable identification method, 11 strains of these yeasts were fully characterized in this study by traditional and advanced technologies. DNA sequences of the internal transcribed spacer (ITS), IGS1, and D1/D2 regions identified six of the yeasts as T. dermatis that were previously known as T. mucoides, including ATCC 204094 that has been used as the quality-control strain of T. mucoides for the VITEK 2 system and other commercial yeast identification kits. These two species could not be differentiated reliably by any previously known phenotypic keys for the species, such as growth patterns on ethylamine, phloroglucinol and tyramine, or by the VITEK 2 system. On the other hand, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) proved to be a rapid and reliable identification tool for the two closely related yeasts. With newly added superspectra from fully authenticated reference strains, the VITEK MS system using MALDI-TOF MS successfully separated strains of T. dermatis and T. mucoides at a similarity level of approximately 67 % for the mass spectra data, and could identify these strains at the species level with 100 % accuracy in repeated tests. Furthermore, the in vitro susceptibility results indicated that itraconazole, posaconazole and voriconazole were more effective against both T. mucoides and T. dermatis than the other antifungal agents tested in this study.
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Comparison of two types of matrix-assisted laser desorption/ionization time-of-flight mass spectrometer for the identification and typing of Clostridium difficile
Microflex LT (Bruker Daltonics) and VITEK MS (bioMérieux) are bacterial identification systems that are based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). For VITEK MS, two identification softwares, VITEK MS IVD (IVD) and SARAMIS (SARAMIS), are available. Microflex LT is equipped with MALDI Biotyper RTC software (Biotyper). Although the identification accuracy of each instrument has been compared for various bacteria, no detailed examination has been conducted for the identification accuracy of Clostridium difficile. In this report, we compared the three identification softwares for identification reproducibility in three ATCC C. difficile strains and identification accuracy in 50 clinical C. difficile isolates. The results showed 100, 91.7 and 100 % identification reproducibility accuracy of ATCC strains when examined by IVD, SARAMIS and Biotyper software, respectively. For the identification of the clinical isolates, all three softwares exhibited satisfactory identification accuracy of C. difficile. Among the 50 clinical isolates, seven showed identical toxin genotype corresponding to the exact ribotype. However, MALDI-TOF MS failed to identify them as the identical type. Based on the above results, we concluded that both types of MALDI-TOF MS reproducibly identified C. difficile; however, they are currently not suitable for typing of C. difficile clones.
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- Antimicrobial Agents and Chemotherapy
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Inhibitory effect of silver nanoparticles mediated by atmospheric pressure air cold plasma jet against dermatophyte fungi
More LessIn an in vitro study with five clinical isolates of dermatophytes, the MIC50 and MIC100 values of silver nanoparticles (AgNPs) ranged from 5 to16 and from 15 to 32 μg ml− 1, respectively. The combined treatment of AgNPs with atmospheric pressure-air cold plasma (APACP) induced a drop in the MIC50 and MIC100 values of AgNPs reaching 3–11 and 12–23 μg ml− 1, respectively, according to the examined species. Epidermophyton floccosum was the most sensitive fungus to AgNPs, while Trichophyton rubrum was the most tolerant. AgNPs induced significant reduction in keratinase activity and an increase in the mycelium permeability that was greater when applied combined with plasma treatment. Scanning electron microscopy showed electroporation of the cell walls and the accumulation of AgNPs on the cell wall and inside the cells, particularly when AgNPs were combined with APACP treatment. An in vivo experiment with dermatophyte-inoculated guinea pigs indicated that the application of AgNPs combined with APACP was more efficacious in healing and suppressing disease symptoms of skin as compared with the application of AgNPs alone. The recovery from the infection reached 91.7 % in the case of Microsporum canis-inoculated guinea pigs treated with 13 μg ml− 1 AgNPs combined with APACP treatment delivered for 2 min. The emission spectra indicated that the efficacy of APACP was mainly due to generation of NO radicals and excited nitrogen molecules. These reactive species interact and block the activity of the fungal spores in vitro and in the skin lesions of the guinea pigs. The results achieved are promising compared with fluconazole as reference antifungal drug.
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In vitro activity of colistin in antimicrobial combination against carbapenem-resistant Acinetobacter baumannii isolated from patients with ventilator-associated pneumonia in Vietnam
Vien Le Minh, Nguyen Thi Khanh Nhu, Voong Vinh Phat, Corinne Thompson, Nguyen Phu Huong Lan, Tran Vu Thieu Nga, Pham Thi Thanh Tam, Ha Thanh Tuyen, Tran Do Hoang Nhu, Nguyen Van Hao, Huynh Thi Loan, Lam Minh Yen, Christopher M. Parry, Ho Dang Trung Nghia, James I. Campbell, Tran Tinh Hien, Louise Thwaites, Guy Thwaites, Nguyen Van Vinh Chau and Stephen BakerAcinetobacter baumannii has become one of the major infection threats in intensive care units (ICUs) globally. Since 2008, A. baumannii has been the leading cause of ventilator-associated pneumonia (VAP) in our ICU at an infectious disease hospital in southern Vietnam. The emergence of this pathogen in our setting is consistent with the persistence of a specific clone exhibiting resistance to carbapenems. Antimicrobial combinations may be a strategy to treat infections caused by these carbapenem-resistant A. baumannii. Therefore, we assessed potential antimicrobial combinations against local carbapenem-resistant A. baumannii by measuring in vitro interactions of colistin with four antimicrobials that are locally certified for treating VAP. We first performed antimicrobial susceptibility testing and multilocus variable number tandem repeat analysis (MLVA) genotyping on 74 A. baumannii isolated from quantitative tracheal aspirates from patients with VAP over an 18-month period. These 74 isolates could be subdivided into 21 main clusters by MLVA and >80 % were resistant to carbapenems. We selected 56 representative isolates for in vitro combination synergy testing. Synergy was observed in four (7 %), seven (13 %), 20 (36 %) and 38 (68 %) isolates with combinations of colistin with ceftazidime, ceftriaxone, imipenem and meropenem, respectively. Notably, more carbapenem-resistant A. baumannii isolates (36/43; 84 %) exhibited synergistic activity with a combination of colistin and meropenem than carbapenem-susceptible A. baumannii isolates (2/13; 15 %) (P = 0.023; Fisher's exact test). Our findings suggest that combinations of colistin and meropenem should be considered when treating carbapenem-resistant A. baumannii infections in Vietnam, and we advocate clinical trials investigating combination therapy for VAP.
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Evaluation of cefazolin as a surrogate marker for cefpodoxime susceptibility for urinary tract isolates
More LessOf the cephalosporins, cefpodoxime has the most published clinical data for the treatment of urinary tract infections. In 2014, the Clinical and Laboratory Standards Institute (CLSI) guidelines recommended that cefazolin should be used as the surrogate marker for cefpodoxime among urinary tract isolates, replacing cephalothin. This study attempted to determine how well cefazolin serves as the surrogate marker. Additionally, it investigated how cefuroxime compared with cefazolin as a surrogate marker. The MicroScan Walkaway Plus system was used to determine susceptibility for cefazolin and cefuroxime on consecutive urine cultures with a colony count of ≥ 50 000 organisms. Only Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis isolates were included, following CLSI guidelines. Simultaneously, an Etest for cefpodoxime was conducted. The cefpodoxime interpretation was compared with that of the other two agents, and the categorical agreement was calculated, defined as the percentage of identical susceptibility interpretations. Cefazolin (92 %) had a significantly higher categorical agreement than cefuroxime (85 %) among 284 isolates (P = 0.011). The major error rate was 4.4 % for cefazolin and 1.1 % for cefuroxime. The very major error rate was 64 % for cefazolin and 18 % for cefuroxime among the 11 cefpodoxime-resistant isolates. Cefazolin was a better predictor of cefpodoxime susceptibility than the previously recommended agent, cephalothin. However, cefuroxime had better major and very major error rates than cefazolin.
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- Clinical Microbiology
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Improving the molecular diagnosis of Chlamydia psittaci and Chlamydia abortus infection with a species-specific duplex real-time PCR
Chlamydia psittaci and Chlamydia abortus are closely related intracellular bacteria exhibiting different tissue tropism that may cause severe but distinct infection in humans. C. psittaci causes psittacosis, a respiratory zoonotic infection transmitted by birds. C. abortus is an abortigenic agent in small ruminants, which can also colonize the human placenta and lead to foetal death and miscarriage. Infections caused by C. psittaci and C. abortus are underestimated mainly due to diagnosis difficulties resulting from their strict intracellular growth. We developed a duplex real-time PCR to detect and distinguish these two bacteria in clinical samples. The first PCR (PCR1) targeted a sequence of the 16S–23S rRNA operon allowing the detection of both C. psittaci and C. abortus. The second PCR (PCR2) targeted the coding DNA sequence CPSIT_0607 unique to C. psittaci. The two PCRs showed 100 % detection for ≥ 10 DNA copies per reaction (1000 copies ml− 1). Using a set of 120 samples, including bacterial reference strains, clinical specimens and infected cell culture material, we monitored 100 % sensitivity and 100 % specificity for the detection of C. psittaci and C. abortus for PCR1. When PCR1 was positive, PCR2 could discriminate C. psittaci from C. abortus with a positive predictive value of 100 % and a negative predictive value of 88 %. In conclusion, this new duplex PCR represents a low-cost and time-saving method with high-throughput potential, expected to improve the routine diagnosis of psittacosis and pregnancy complication in large-scale screening programs and also during outbreaks.
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Evolution of macrolide resistance in Streptococcus pyogenes over 14 years in an area of central Italy
We evaluated temporal fluctuations in macrolide resistance rates, analysing genetic determinants of resistance and clonal evolution in a population of 2744 S. pyogenes isolates collected in the period 2000–2013. The total resistance rate to erythromycin of the isolates was 17.9 %. A maximum of erythromycin resistance emerged in 2000 (38.6 %), followed by a significant decrease to 5.2 % in 2012 (P < 0.0001). Molecular analysis revealed the presence and co-presence of known genetic resistance determinants mefA, mefE, ermTR and ermB, in line with phenotypes. PFGE analysis identified genetically related groups in 2000 and 2007–2008, mainly the MLS and M phenotypes, respectively. The most prevalent emm types among a representative subset of resistant isolates were emm2, emm75 and emm77. All emm2 and 88.2 % of the strains harbouring the emm75 gene were only recorded in M-phenotype strains, whilst all emm77-positive strains had the inducible MLS phenotype. The analysed susceptible isolates showed several emm types partially shared with resistant ones. Our results suggest that changes in bacterial population clonality, rather than horizontal transfer of resistance determinants, plays a major epidemiological role in S. pyogenes. Continuous monitoring of microbiological epidemiology seems to be crucial for correct and effective management of streptococcal infections.
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In vitro activity of minocycline alone or in combination in multidrug-resistant Acinetobacter baumannii isolates
More LessMinocycline (MIN) usually shows good activity against Acinetobacter baumannii strains. The reintroduction to the market of intravenous MIN provides an additional agent to the limited options for the treatment of A. baumannii infections. The activity of MIN as a single agent and in combination with rifampicin (RIF), colistin (COL) or imipenem (IMI) was evaluated by means of killing curves and 24 h-time-kill curves in five A. baumannii isolates which were selected on the basis of different antimicrobial resistance profiles. MIN showed bacteriostatic activity in three isolates (2 × or 16 × MIC) and bactericidal activity in the other isolates (64 × MIC). In isolates harbouring the tetB gene, the associations studied were always indifferent. However, in isolates not harbouring tetB, the use of MIN in combination showed a rapid synergistic effect (at 4 h) in four out of nine combinations (two with RIF and one each with IMI and COL). At 24 h, this effect was observed in six out of nine combinations (two in each association). MIN in combination with RIF, IMI and COL showed bactericidal synergy in most of the isolates which did not harbour the tetB gene, but the combinations were not synergistic in tetB-positive isolates.
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Human toxoplasmosis: a comparative evaluation of the diagnostic potential of recombinant Toxoplasma gondii ROP5 and ROP18 antigens
More LessToxoplasmosis is one of the most common parasitic diseases worldwide and it poses a serious challenge regarding prevention, diagnosis and therapy. The commonly used diagnostic methods are mostly based on the detection of specific antibodies in sera. Since they are not always accurate enough and do not allow precise definition of the phase of the Toxoplasma gondii infection, there is an urgent need to find specific molecular markers of acute or chronic infection stages. This study provides a comparative assessment of recombinant ROP5 and ROP18 T. gondii proteins in the serodiagnosis of human toxoplasmosis. We found that both ROP5 and ROP18 proteins allowed the detection of specific IgM and IgG antibodies with a relatively low sensitivity; however, ROP18 IgM ELISA proved to be more sensitive than the SAG1 assay. This study also points to a relatively weak potential of the corresponding native ROP5 and ROP18 kinases in the generation of a strong antibody response in humans.
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- Microbial Epidemiology
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Comparative analysis of virulence determinants, antibiotic susceptibility patterns and serogrouping of atypical enteropathogenic Escherichia coli versus typical enteropathogenic E. coli in India
The epidemiology of enteropathogenic Escherichia coli (EPEC) and the significance of isolation of atypical EPEC (aEPEC) in childhood diarrhoea have not been well studied in an Indian context. A comparative study was undertaken to investigate virulence determinants, antibiotic susceptibility patterns and serogrouping of typical EPEC (tEPEC) versus aEPEC causing diarrhoea in children. A total of 400 prospective and 500 retrospective E. coli isolates were included. PCR was performed for eae, bfpA, efa, nleB, nleE, cdt, ehxA and paa genes. The Clinical and Laboratory Standards Institute's disc diffusion test was used to determine the antimicrobial susceptibility. Phenotypic screening of extended spectrum β-lactamases (ESBLs), AmpC and Klebsiella pneumoniae carbapenemase (KPC) production, and molecular detection of bla NDM-1, bla VIM, bla CTX-M-15, bla IMP and bla KPC were performed. aEPEC (57.6 %) were more common as compared with tEPEC (42.3 %). The occurrence of virulence genes was observed to be three times higher in aEPEC as compared with tEPEC, efa1 (14.7 % of aEPEC, 4 % of tEPEC) being the most common. Most of the isolates did not belong to the classical EPEC O-serogroups. The highest resistance was observed against amoxicillin (93.22 %) followed by quinolones (83 %), cephalosporins (37.28 %), cotrimoxazole (35.59 %) and carbapenems (30.5 %). Overall equal numbers of aEPEC (41.17 %) and tEPEC (40 %) were observed to be multidrug-resistant. Fifteen EPEC strains demonstrated presence of ESBLs, five produced AmpC and four each produced metallo-β-lactamases and KPC-type carbapenemases; eight, seven and one isolate(s) each were positive for bla VIM, bla CTX-M-15 and bla NDM-1, respectively. Here, to the best of our knowledge, we report for the first time on carbapenem resistance and the presence of bla NDM-1 and bla CTX-M-15 in EPEC isolates from India.
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Molecular identification of clinical Nocardia isolates from India
The epidemiology of nocardiosis is evolving with increasing number of Nocardia spp. causing human infection. In recent years, molecular techniques have been used to identify Nocardia spp. There are limited data available on the spectrum of Nocardia spp. isolated from clinical samples in India. Here, a molecular study was carried on 30 clinical isolates maintained in our National Culture Collection to evaluate the techniques used for identifying the agents. The isolates were identified by sequencing two promising genes: the 16S rRNA gene and hsp65. Both hsp65 and the 16S rRNA gene could reliably identify 90 % of Nocardia isolates, i.e. N. farcinica, N. cyriacigeorgica, N. brasiliensis, N. otitidiscaviarum, N. amamiensis and N. pneumoniae. The mean percentage dissimilarity of sequence identification was higher using the hsp65 gene (4 %, range 0–7.9 %) compared with the 16S rRNA gene (2.3 %, range 0–8.9 %). Two isolates that showed ambiguous results in both the short segment of the 16S rRNA gene and hsp65 sequences could be resolved by sequencing a larger fragment (∼1000 bp) of the 16S rRNA gene. Both of these isolates were identified as N. beijingensis with similarities of 99.8 and 100 % compared with the standard strain. Genotyping of N. cyriacigeorgica strains was performed using hsp65 gene sequences and compared with previously described genotypes. Our N. cyriacigeorgica isolates belonged to genotype 1 (n = 4) and genotype 2 (n = 2). The present study highlights a wide spectrum of Nocardia spp. in India and emphasizes the need for molecular techniques for identification to the species level.
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Predominance of PCR-ribotypes, 018 (smz) and 369 (trf) of Clostridium difficile in Japan: a potential relationship with other global circulating strains?
Mitsutoshi Senoh, Haru Kato, Tadashi Fukuda, Akiko Niikawa, Yoshiko Hori, Hideharu Hagiya, Yoichiro Ito, Hiroshi Miki, Yoshifumi Abe, Kiyoshi Furuta, Hideki Takeuchi, Hirokazu Tajima, Harumi Tominaga, Hideyuki Satomura, Hideaki Kato, Sayuri Morita, Ai Tanada, Toshinori Hara, Miki Kawada, Yuka Sato, Masahiko Takahashi, Akiko Higuchi, Tomoko Nakajima, Yukiko Wakamatsu, Masahiro Toyokawa, Akiko Ueda, Paul Roberts, Fabio Miyajima and Keigo ShibayamaGlobal spread and evolutionary links of an epidemic Clostridium difficile strain (PCR-ribotype 027) have been noted in recent decades. However, in Japan, no outbreaks caused by type 027 have been reported to date. A total of 120 C. difficile isolates from patients at 15 hospitals during non-outbreak seasons between 2011 and 2013 as well as 18 and 21 isolates collected from two hospitals in 2010 and 2009, respectively, in outbreak periods in Japan, were examined. Among these 120 isolates, Japan-ribotypes smz and ysmz (subtype variant of smz) were the most predominant (39.2 %) followed by Japan-ribotype trf (15.8 %). Types smz/ysmz and trf were also concurrently predominant at two hospitals in the outbreak settings. Out of the five binary toxin-positive isolates observed, only one was PCR-ribotype 027 and another PCR-ribotype 078. Type smz was later found to correspond to PCR-ribotype 018. High rates of resistance against gatifloxacin, moxifloxacin, erythromycin and clindamycin were observed in the PCR-ribotype 018 isolates. Interestingly, all trf isolates were toxin A-negative, toxin B-positive, but they did not correspond to PCR-ribotype 017, thus being assigned a new ribotype (PCR-ribotype 369). In conclusion, PCR-ribotypes 018 (smz) and 369 (trf) were identified as major circulating strains in both outbreak and non-outbreak settings in Japan. Given their epidemiological relevance, molecular investigations are warranted to clarify potential evolutionary links with related strains found elsewhere, such as PCR-ribotypes 018 and 017 from Europe and North America.
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- Models of Infection
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Whole glucan particles as a vaccine against systemic coccidioidomycosis
We reported previously that yeast-derived whole glucan particles (WGPs), with or without conjugation to BSA, used as a vaccine protected against systemic aspergillosis in mice. Here, we examined their utility as a potential vaccine against coccidioidomycosis. WGPs were prepared from Saccharomyces cerevisiae; conjugation with BSA (WGP–BSA) was done using 1-cyano-4-dimethylaminopyridinium tetrafluoroborate-mediated conjugation. Heat-killed S. cerevisiae (HKY) was used as a positive-control vaccine. CD-1 mice were vaccinated with WGPs or WGP–BSA, HKY or PBS once weekly, beginning 21 days prior to infection. Mice were infected intravenously with arthroconidia of Coccidioides posadasii. In the low-mortality study, 50 % of PBS-treated controls died. Only WGP–BSA at 0.6 mg per dose induced significant protection compared with PBS treatment. All surviving mice were infected in all three organs examined. Those given WGP–BSA at 0.6 mg per dose had fewer c.f.u. in liver and lungs (P = 0.04), and those given WGPs at 6 mg per dose had fewer in lungs (P < 0.02), compared with PBS. In the high-mortality study, 90 % of PBS mice died. Vaccination with HKY, and WGPs or WGP–BSA at 6 or 12 mg per dose significantly prolonged survival (P ≤ 0.05). No surviving mice were free of infection. HKY and WGP–BSA at 12 mg per dose reduced c.f.u. in the liver and lungs (P < 0.05) and WGP–BSA at 6 mg per dose reduced c.f.u. in the lungs (P < 0.05); unconjugated WGPs did not reduce infection. WGPs or WGP–BSA acted as a vaccine that protected against mortality caused by coccidioidomycosis. Thus, WGP protection against coccidioidomycosis and aspergillosis provides the basis for development of a pan-fungal vaccine.
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- Prevention and Therapy
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Inactivating effects of the lactoperoxidase system on bacterial lyases involved in oral malodour production
The main components of oral malodour have been identified as volatile sulfur compounds (VSCs), including hydrogen sulfide (H2S) and methyl mercaptan (CH3SH). The lactoperoxidase (LPO) system (consisting of LPO, glucose oxidase, glucose and thiocyanate) was previously shown to exhibit antimicrobial activities against some oral bacteria in vitro and suppressive effects on VSCs in mouth air in a clinical trial. Here, we examined the in vitro effects of the LPO system on the activities of the bacterial lyases involved in the production of VSCs by oral anaerobes. The exposure of crude bacterial extracts of Fusobacterium nucleatum and Porphyromonas gingivalis or purified methionine γ-lyase to the LPO system resulted in the inactivation of their lyase activities through l-cysteine and l-methionine, which was linked to the production of H2S and CH3SH, respectively. The exposure of living F. nucleatum and P. gingivalis cells to the LPO system resulted in the suppression of cell numbers and lyase activities. The inactivation of the crude bacterial extracts of F. nucleatum and purified methionine γ-lyase by the LPO system was partly recovered by the addition of DTT. Therefore, the LPO system may inactivate bacterial lyases including methionine γ-lyase by reacting with the free cysteine residues of lyases. These results suggested that the LPO system suppresses the production of VSCs not only through its antimicrobial effects, but also by its inactivating effects on the bacterial lyases of F. nucleatum and P. gingivalis.
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Volumes and issues
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Volume 73 (2024)
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Volume 72 (2023 - 2024)
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Volume 71 (2022)
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Volume 70 (2021)
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Volume 69 (2020)
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Volume 68 (2019)
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Volume 67 (2018)
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Volume 65 (2016)
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Volume 1 (1968)