The aim of this study was to determine and compare thymic output during 12 months of highly active antiretroviral therapy (HAART) in HIV-infected patients with different types of immune recovery. In total, 18 Thai HIV-infected patients with normal immune recovery (NR) and 13 Thai HIV-infected patients with slow immune recovery (SR) were enrolled. T-cell receptor rearrangement excision circle (TREC) levels in peripheral blood mononuclear cells (PBMCs) and CD4⁺ T cells were quantified at baseline, and after 6 and 12 months of HAART. CD4⁺ T-cell counts in NR patients were significantly higher than those in SR patients after 6 and 12 months of HAART. However, the median TREC levels in PBMCs and CD4⁺ T cells in both groups were comparable. Moreover, TREC levels showed similar trends in PBMCs and CD4⁺ T cells in both groups during 12 months of HAART. Only patients with SR had significant increases in median TREC levels in PBMCs and CD4⁺ T-cells during the first 6 months of HAART. No correlations were found between CD4⁺ T-cell count and TREC levels in PBMCs and CD4⁺ T cells. These results imply that the increase in CD4⁺ T-cell count in SR patients after 12 months of HAART is likely attributable to thymic output and other sources.

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The development of a dengue virus vaccine is a major priority in efforts to control the diseases. Several researchers are currently using the Asian 1 and Asian 2 genotypes as vaccine candidates for dengue type 2 virus (DENV-2). However, in this study, we constructed a recombinant plasmid-based prM/E gene, from a DENV-2 Cosmopolitan genotype strain as a dengue DNA vaccine candidate. The protein expression of the recombinant plasmid in CHO cells was analyzed using an enzyme-linked immunosorbent assay, western blotting, and sucrose gradient sedimentation. After being used to immunize ddY mice three times at doses of 25 or 100 μg, the DNA vaccine induced humoral immune responses. There was no difference in the neutralizing antibody titer (focus reduction neutralization test 50% value) of mice immunized with 25 and 100 μg DNA vaccine doses. When challenged with 3 × 10⁵ FFU DENV-2, immunized mice could raise anamnestic neutralizing antibody responses, which were observed at day 4 and day 8 post-challenge. Analysis of immunogenicity using BALB/c mice showed that their antibody neutralization titers were lower than those of ddY mice. In addition, the antibodies produced after immunization and challenge could also neutralize a DENV-2 Asian 2 genotype (New Guinea C) strain. Therefore, the DENV-2 Cosmopolitan genotype may be a DENV-2 vaccine candidate.

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Infection with the novel H10N8 virus in humans has raised concerns about its pandemic potential worldwide. We report the results of a cross-sectional study of avian influenza viruses (AIVs) in live poultry markets (LPMs) in Nanchang, China, after the first human case of H10N8 virus infection was reported in the city. A total of 201 specimens tested positive for AIVs among 618 samples collected from 24 LPMs in Nanchang from December 2013 to January 2014. We found that the LPMs were heavily contaminated by AIVs, with H9, H10, and H5 being the predominant subtypes and more than half of the LPMs providing samples that were positive for the H10 subtype. Moreover, the coexistence of different subtypes was common in LPMs. Of the 201 positive samples, 20.9% (42/201) had mixed infections with AIVs of different HA subtypes. Of the 42 mixed infections, 50% (21/42) showed the coexistence of the H9 and H10 subtypes, with or without H5, and were from chicken samples. This indicated that the H10N8 virus probably originated from segment reassortment of the H9 and H10 subtypes.

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We aimed to assess the risk of infections caused by sexual practices and determine the factors affecting the prevalence of human immunodeficiency virus (HIV) and syphilis among men who have sex with men (MSM) in Lanzhou, China. A survey including 600 MSM was conducted from March 2008 to December 2009. Demographic and sexual behavior data were collected using the snowball sampling technology, and blood samples were obtained. Multivariate logistic regression models were used to determine the predictors of HIV and syphilis infection. The seroprevalence of HIV, syphilis, HCV, and co-infection of HIV with syphilis was 6.0%, 18.0%, 1.3%, and 2.5%, respectively. Use of condoms was more inconsistent in vaginal sex than that in anal sex, buying sex or selling sex. Multivariate analysis showed that MSM with syphilis infection, MSM from other provinces, and MSM of Han ethnicity were more likely to have HIV infection. Further, MSM with HIV infection, inconsistent condom use, and less knowledge about AIDS prevention were more likely to have syphilis infection. Their social background, habit of condom use, and infection status may affect the prevalence of HIV or syphilis. These findings may have important implications for further behavioral interventions among this population.

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The prevention and control of methicillin-resistant Staphylococcus aureus (MRSA) are important, particularly in emergency units. The active surveillance of MRSA was prospectively performed at the emergency medical center of Nagasaki University Hospital. After obtaining nasal swab specimens, a fully automated molecular test (FAMT) and a culture-screening method were utilized for MRSA detection. A total of 150 patients were enrolled in the study, and 366 nasal swab specimens were obtained. MRSA was detected by culture in 11 (7.3%) patients including one new acquisition and by the FAMT in 34 (22.7%) patients including 13 new acquisitions. The sensitivity, specificity, positive predictive value, and negative predictive value of the FAMT at the patient level were 86.7, 85.2, 39.4, and 98.3%, respectively, when compared with the culture-based results. An FAMT can effectively detect MRSA colonization, which may remain undetected with the conventional method, and it may be useful in detecting newly acquired MRSAs.

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Health care-associated infections (HCAIs) cause considerable morbidity and mortality in pediatric intensive care units (PICUs). The objective of this point prevalence study was to assess the burden of HCAIs in PICUs in Turkey. Fifty PICUs participated in this study. Data regarding demographics, microbiological findings, therapeutic interventions, and outcomes were collected for all PICU inpatients. A total of 327 patients participated in the study: 122 (37%) experienced 1 or more HCAI. The most frequently reported site of infection was lower respiratory tract (n=77, 63%). The most frequently isolated pathogens were Pseudomonas aeruginosa, Acinetobacter species, and Candida species. Two hundred and forty-seven patients (75%) were receiving antimicrobial therapy at the time of the survey, and the most frequently administered antimicrobials were third generation cephalosporins. Hospital type, male, PICU stay >7 days, and mechanical ventilation were found to be independent risk factors for HCAIs. At the 4-week follow up, 43 (13%) patients had died, 28 (65%) of whom died of HCAIs. Endotracheal intubation, urinary catheter, male, and HCAIs were independent risk factors for mortality. This national, multicenter study documented a high prevalence of HCAIs in Turkey. In light of the ‘primum non nocere’ principle, the prevention of these infections should be a priority of public health policy.

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In this study, G proteins of the rabies virus (RABV) Kyoto strain were detected in the cytoplasm but not distributed at the cell membrane of mouse neuroblastoma (MNA) cells. G proteins of CVS-26 were detected in both the cell membrane and perinuclear space of MNA cells. We found that N-glycosylation of street RABV G protein by the insertion of the sequon Asn²⁰⁴ induced the transfer of RABV G proteins to the cell surface membrane. Fixed RABV budding from the plasma membrane has been found to depend not only on G protein but also on other structural proteins such as M protein. However, the differing N-glycosylation of G protein could be associated with the distinct budding and antigenic features of RABV in street and fixed viruses. Our study of the association of N-glycan of G protein at Asn²⁰⁴ with the transport of RABV G protein to the cell surface membrane contributes to the understanding of the evolution of fixed virus from street virus, which in turn would help for determine the mechanism underlying RABV budding and enhanced host immune responses.

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The frequency distributions of the numbers of patients and eaters were near lognormal for almost all food poisonings. The medians and ranges remained almost unchanged over time, although the annual number of incidents varied. However, the numbers of patients and eaters were not correlated for many of the food poisonings. A regular pattern relating the numbers of patients and eaters emerged for food poisonings exhibited strong seasonal incidence. For example, food poisonings caused by norovirus or Campylobacter spp. exhibited a comb teeth-like pattern in the plot comparing the ranking numbers of patients and that of eaters, and fingerprint-like pattern in the plots comparing the attack rate and number of patients or eaters. Meanwhile, food poisonings with low incidence and without seasonality, such as those caused by Clostridium perfringens, exhibited amorphous patterns in both plots. Further analysis indicated the occurrence of these patterns was determined by the combination of pathogens and services, and not by either factor alone. Thus, the emergence of a regular pattern may be associated with “complexity” nature of food poisoning.

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Acute Toxoplasma gondii infection causes different clinical courses in 10–20% of cases. In immunocompetent patients, toxoplasmosis most often presents as asymptomatic cervical lymphadenopathy. Clinical manifestations such as pneumonia, myocarditis, hepatitis, and encephalitis are rarely reported. We present the case of an immunocompetent patient with a serious and complicated clinical course of toxoplasmic hepatitis with a maculopapular rash. The diagnosis was confirmed by serology and identification of bradyzoites in liver biopsy samples.

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Chikungunya (CHIK) is a mosquito-borne viral disease. In the 2004 CHIK outbreak in Kenya, diagnosis was delayed because of the lack of accurate diagnostics. Therefore, this study aimed to develop and evaluate an in-house IgM-capture enzyme linked immunosorbent assay (ELISA) (in-house ELISA) for the detection of chikungunya virus (CHIKV) infections. Anti-CHIKV antibodies were raised in rabbits, purified and conjugated to horseradish peroxidase. These anti-CHIKV antibodies and cell-culture derived antigen were used to develop the ELISA. To validate the in-house ELISA, 148 patient sera from the 2005 Comoros CHIK outbreak were tested with centers for disease control and prevention (CDC) IgM-capture ELISA (CDC ELISA) and focus reduction neutralization test (FRNT) as reference assays. The in-house ELISA had a sensitivity of 97.6% and specificity of 81.3% compared to the CDC ELISA and a sensitivity of 91.1% and specificity of 96.7% compared to FRNT. Furthermore, 254 clinically suspected dengue patient samples from Eastern Kenya, collected in 2013, were tested for CHIKV IgM using the in-house ELISA. Out of the 254 samples, 26 (10.2%) were IgM positive, and of these 26 samples, 17 were further analyzed by FRNT and 14 (82.4%) were positive. The in-house ELISA was able to diagnose CHIKV infection among suspected dengue cases in the 2013 outbreak.

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In this study, we investigated whether Vibrio cholerae cells exposed to nutrient-limited conditions developed resistance to abiotic stresses because of which most, if not all, starved cells turn coccoid in shape and exhibited marked resistance to freezing, low pH, and chlorine, but not against bile.

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The regulatory mechanism of miRNA induction in response to Mycobacterium tuberculosis (MTB) infection has not been clearly established. Autophagy has recently been identified as an effective way to control intracellular survival of MTB. In the present study, we demonstrate a novel role of miR-30A in the negative regulation of the autophagy-mediated anti-MTB response. We found that overexpression of miR-30A suppresses the elimination of intracellular MTB through the inhibition of autophagy. Furthermore, there was a negative correlation between concentrations of miR-30A and beclin-1 in MTB positive patients and miR-30A expression decreased after anti-TB treatment. Our results indicate that miR-30A plays a key role in immune response against MTB and, therefore, may serve as a potential target for future treatments of tuberculosis infection.

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Dengue fever, an acute, mosquito-borne, febrile illness caused by Flavivirus spp., is a problem in Africa, South and Southeast Asia, Latin America, and the Caribbean. A dengue outbreak occurred after nearly 70 years of absence or no detection, and then 158 autochthonous cases occurred in Japan from August to October 15, 2014. The most competent mosquito vectors for dengue virus transmission were Aedes aegypti and A. albopictus. Since A. albopictus is widely distributed across Japan and A. aegypti recently invaded Japan by airplane, we examined the susceptibility of these species to infection by dengue virus.

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The aim of this study was to determine the resistance patterns of uropathogenic Escherichia coli (UPEC) isolates and to investigate the frequency of several virulence genes, including fimH, papA, hlyD, cnf-1, sitA, and tsh, among various phylogenetic groups of UPEC isolates. A total of 85 E. coli isolates were recovered from urine samples from outpatients with a clinical diagnosis of uncomplicated urinary tract infections. A molecular approach to examine the antimicrobial resistance patterns was employed using PCR and the disc diffusion method. The detected frequencies of the virulence factor genes determined using PCR were: fimH (34.1%), papA (9.4%), hlyD (21.2%), cnf-1 (3.5%), sitA (15.3%), and tsh (27.1%). These results revealed that the isolates were resistant to trimethoprim-sulfamethoxazole (SXT) (74.1%), cefotaxime (CTX) (68.2%), and amoxicillin-clavulanic acid (AMC) (94.1%), and they were relatively less resistant to N (56.5%). According to these results, further investigation is needed to determine exactly whether or not SXT, CTX, and AMC are appropriate antibiotics for the treatment of UPEC infections in southern Iran. Although these results demonstrate that fimH is the most frequent virulence gene among UPEC isolates, the high prevalence of isolates that do not encode fimH (75.9%) and the relatively low frequency of isolates that carry other virulence genes require further investigation to clarify the role of the other potential virulence factors in the pathogenesis of these isolates.

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Campylobacter species are common causes of bacterial diarrhea, and Campylobacter jejuni and C. coli are known as the predominant causative agents in humans. Recent studies suggested that loop-mediated isothermal amplification (LAMP) is an efficient and practical tool for rapid detection of C. jejuni and C. coli in clinical samples. We used LAMP to screen 151 stool samples for Campylobacter; these samples were collected in 2012 from Thai children with diarrhea. The PCR method discriminated C. jejuni and C. coli among the detected Campylobacter strains; these species were subjected to sequencing of the hipO gene (in C. jejuni) or the ask gene (in C. coli). The results suggest that the prevalence of Campylobacter infection among Thai children with diarrhea is 8.6%, and C. jejuni is the most prevalent species.

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We surveyed Rickettsiales bacteria, including Rickettsia, Ehrlichia, Anaplasma, and Neoehrlichia, in wild sika deer (Cervus nippon nippon) from Shizuoka prefecture, Japan. In spleen samples from 187 deer, Anaplasma phagocytophilum (deer type), A. bovis, and A. centrale were successfully detected by PCR assay targeting to 16S rDNA or p44/msp2, and their positive rates were 96.3% (180/187), 53.5% (100/187), and 78.1% (146/187), respectively. Additionally, 2 or 3 Anaplasma species could be detected from a single deer in 165 spleen samples (88.2%), indicating dual or triple infection. In contrast, A. phagocytophilum (human type) 16S rDNA, Rickettsia gltA, Ehrlichia p28/omp-1, and Neoehrlichia 16S rDNA could not be amplified. The serological test of 105 deer serum samples by immunofluorescence assay showed that the detection of antibodies against antigens of A. phagocytophilum HZ (US-human isolate) and Rickettsia japonica YH were 29.5% (31/105) and 75.2% (79/105), respectively. These findings suggest that A. phagocytophilum (deer type), A. centrale, and A. bovis are highly dominant and prevalent in wild sika deer from Shizuoka, a central region of Japan, and that the antibodies against some Rickettsiales bacteria have also been retained in deer blood.

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