Candida is the most frequently encountered fungal infection of the gastrointestinal tract after antibiotic exposure. The pathogenesis of Candida probably varies with each species. The speciation of fecal Candida after antibiotic use is not well investigated. One hundred and eleven fecal samples negative for Clostridium difficile toxin and for other enteric pathogens formed the basis of our investigation. The diarrheic samples came from patients receiving antibiotics in a hospital setting. In addition, samples from 30 age-matched healthy participants who did not receive antibiotics and did not have diarrhea were also studied. Initially, a Gram stain identification for yeasts was performed for each fecal sample, then each sample was cultured on Sabouraud's dextrose agar. Candida was isolated as pure growth (>10⁵ cfu/ml) from the stools of 32 (28.8%) patients. The identification of the yeast was done based upon a combination of morphological, physiological and biochemical criteria. The predominant isolates were C. tropicalis (n = 16), C. albicans (n = 14) and C. krusei (n=2). Candida isolated from healthy participants (n = 4) was sparse and therefore not speciated. Different Candida spp. may play an important role in precipitating antibiotic-associated diarrhea.

The hospital records of 213 outpatients from Bangkok, Thailand, infected with Strongyloides stercoralis as determined by stool inspections were examined retrospectively for the different clinical presentations ascribed to patients with HIV, those with chronic illness, those who used immunosuppressant drugs and relatively healthy subjects. For HIV patients with strongyloidiasis, the most common symptoms were chronic diarrhea, fever, persistent coughing and loss of weight, but only the first three symptoms were significantly different from other immunocompromised hosts. For healthy patients with strongyloidiasis, acute diarrhea and abdominal pain were the most frequent symptoms. Moreover, the peripheral eosinophil blood count was significantly lower (P = 0.004) in the HIV patients than in any of the other subsets. Males were more common than females across all categories. While the average age of all subjects was 48.3 ± 16.4 years, the strongyloidiasis patients with chronic illness were significantly older (56.8 ± 13.5 years) than those in the other groups. This study may suggest that strongyloidiasis is commonly found in geriatric males, and that the patients most at risk for S. stercoralis infection are HIV patients. This is the first report of the different clinical presentations of intestinal strongyloidiasis in various groups of patients with impaired immunity.

Mouse 3T3-Swiss albino cells are widely used as feeder cells to culture the human epidermis for the treatment of burns. To minimize the risk of xenoinfection, quality control of the feeder cells is required in the Japanese guidelines on regenerative medicine using feeder cells. We characterized three lots of 3T3-Swiss albino cells that are publicly or commercially available in Japan. One lot, which propagated more rapidly than the other two without showing typical contact inhibition, was found to release endogenous murine leukemia virus upon iododeoxyuridine-treatment. Southern blotting of restriction fragments showed that the rapidly growing lot consisted of genetically altered cells that had probably emerged during the passages. The data support the guidelines that recommend the quality control of each lot of 3T3-Swiss albino cells if they are to be used clinically.

Quinolone resistance mostly originates from chromosomal mutations. In recent years, however, plasmid-mediated quinolone resistance has been reported in several parts of the world. Plasmid-borne qnrA, qnrB, or qnrS genes are responsible for this kind of resistance. Little is known about the diversity, type, and species range of the qnr genes in Turkey. We screened qnrA, qnrB, and qnrS genes in quinolone-resistant blood culture isolates collected from six different medical centers in Turkey which produced extended-spectrum β-lactamases (ESBLs). A total of 78 ESBL-positive isolates were enrolled in this study. Of these, 37 (47.4%) were nalidixic-acid resistant or intermediate. qnrA was found on large plasmids isolated from five (6.4%) of the Nal^I/R isolates. In three of these, the same plasmid also carried bla_CTX-M. Four of the qnrA-positive isolates were Klebsiella pneumoniae from Dokuz Eylul University Hospital, Izmir, and the fifth isolate was Escherichia coli from Istanbul University Hospital. Two of the isolates from Izmir were found by enterobacterial repetitive interegenic consensus sequence-PCR to be clonally related. This is the first report on the qnrA prevalence among ESBL-positive blood culture isolates collected from different regions in Turkey. According to our results, plasmid-mediated resistance is a potential problem for the spread of quinolone resistance, and this mechanism could be emerging strongly among the ESBL-positive Enterobacteriaceae in Turkey.

Although polymerase chain reaction (PCR) is a highly sensitive procedure for the diagnosis of enteroviruses, it has never been systemically applied to the treatment of enteroviral encephalitis using intravenous immunoglobulin (IVIg). We conducted a 2-year randomized, controlled comparison of reverse transcription (RT)-PCR of cerebrospinal fluid (CSF) with traditional viral isolation to guide IVIg treatment. Seventy-five patients were enrolled and classified into three groups: one group with clinical manifestations of enteroviral infections and two without. The latter two groups were separated on the basis of whether IVIg treatment was guided by RT-PCR or virus culture assay. CSF specimens from the 18 confirmed cases of enteroviral encephalitis were RT-PCR positive for enterovirus in all but one case. Of the remaining 57 cases of nonenteroviral encephalitis, only 4 were positive for enterovirus RT-PCR. One patient in the group of IVIg treatment guided by viral isolation subsequently displayed a sequel of epilepsy. No patients in the IVIg treatment groups guided by RT-PCR had any neurological sequelae. In conclusion, the use of RT-PCR allowed rapid, sensitive, and specific detection of enteroviral RNA in CSF. When used to guide IVIg treatment, RT-PCR may shorten hospitalization and improve outcomes of patients with enteroviral encephalitis.

We aimed to evaluate the treatment outcome of pulmonary tuberculosis patients and factors affecting treatment outcomes. We analyzed the records of 586 pulmonary tuberculosis patients who were older than 15 years followed between January 1999 and December 2004. Of these patients, 76.1% were smear-positive for tuberculosis and 23.9% were smear-negative for tuberculosis. The treatment outcomes of all patients analyzed were as follows: treatment success 91.7%, defaulted treatment 5.1%, died 2.4%, failure 0.3%, and transferred out 0.5%. The treatment outcomes of smear-positive pulmonary tuberculosis patients were as follows: cured 77.1%, treatment completed 13.5%, defaulted treatment 5.4%, died 2.9%, failure 0.4%, and transferred out 0.7%. In multivariate regression analysis, risk factors for non-successful treatment outcome were determined to be re-treatment patients, patients older than 46 years of age, and the presence of rifampicin resistance. We conclude that application of Directly Observed Therapy may increase treatment success in all patients, especially patients who have risk factors for a low treatment success rate.

Traditional risk factors are not adequate for predicting serious bacterial infection (SBI) in febrile infants. The purpose of this study was to evaluate the diagnostic value of the plasma level of soluble form of triggering receptor expressed on myeloid cells-1 (sTREM-1) in predicting SBI in febrile infants less than 3 months old. Forty-four febrile infants less than 3 months old with clinical suspicion of SBI were enrolled. Blood was drawn for measurement of plasma sTREM-1 levels, and microbiological cultures were obtained at the time of admission. Twenty-three infants (52.3%) had SBI and 21 infants (47.7%) had no evidence of SBI based on the results of bacterial culture. sTREM-1 levels were significantly higher in infants with SBI than in infants without SBI (mean ± SD, 324.6 ± 546.3 versus 7.7 ± 16.4, P < 0.0001 after adjusting for age). The area under the receiver-operating characteristic curve was 0.88 for the sTREM-1 level. At a cutoff level of 24.4 ng/mL, the sTREM-1 level yielded a sensitivity of 87%, a specificity of 81%, a positive likelihood ratio of 4.6, and a negative likelihood ratio of 0.2 for differentiating between presence and absence of SBI. In conclusion, sTREM-1 may become a valuable diagnostic tool in the initial evaluation of febrile young infants.

Pneumonia is the leading cause of pediatric morbidity and mortality worldwide, and Mycoplasma pneumoniae and Chlamydia pneumoniae are the two most common atypical pathogens. This study was designed to determine the prevalence and clinical impact of mycoplasma and chlamydia pneumonia in children hospitalized with severe pneumonia. Children 1 month - 15 years old with a diagnosis of severe pneumonia (WHO criteria) were recruited between March 2005 and March 2006. Serologic studies were performed for anti-M. pneumoniae and anti-C. pneumoniae IgG/M on admission and 2-4 weeks afterward using ELISA. Of 52 patients, 13 (25%) were positive for Mycoplasma, 8 (15%) were positive for Chlamydia, 4 (7.6%) were positive for a mixed infection and 27 (52%) were negative. The subjects' mean age was 23.8 ± 4.1 months. The mean of initial oxygen saturation on admission was 87.5 ± 1.2%. Fever and prolonged cough were the leading symptoms. The mean of hospitalization was 18.8 ± 2.6 days, chlamydia pneumonia had the longest duration, 30 ± 10.2 days and 13/52 (25%) study subjects developed respiratory failure. Only 10% were treated with adequate antibiotic prior to serologic results. There was one mortality (1/52, 2%). Our study suggests that mycoplasma and chlamydia infections are commonly found among children hospitalized with severe pneumonia. Coverage with an appropriate antibiotic should be considered to hasten recovery.

Japanese encephalitis is a severe central nervous system (CNS) disease with a high case fatality rate in humans. We characterized T-cells infiltrating the brain after infection with Japanese encephalitis virus (JEV) in a mouse model and determined the clonality of the infiltrating T-cells by analyzing the sequences of complementary determining region 3 (CDR3) of the T-cell receptor. C3H/He mice died after intraperitoneal infection with the JaTH160 strain of JEV, demonstrating CNS degeneration and prominent T-cell infiltration. The percentages of T-cells bearing the VA5-1, VA17-1, VA19-1, VB2-1, VB8-3 and VB13-1 subfamilies were significantly increased following infiltration of the brains in infected mice. Additionally, CDR3 size spectratyping revealed the oligoclonality in T-cells bearing VA11-1 and VA18-1. CDR3 amino acid sequences were then determined for the VA5-1, VA11-1, VA18-1, VB8-3 and VB13-1 subfamilies. There were high levels of identity and similarity in amino acid sequences of CDR3 in these T-cells. Quantitative real-time PCR analysis also revealed that CD8, interferon-γ and tumor necrosis factor-α were highly expressed in the infected mouse brain. These results indicate that T-cells with high clonality and similarity infiltrate the JEV-infected mouse brain, and that these T-cells are mainly CD8-positive and have the Th1/Tc1 phenotype.

In this report we describe the clinical and laboratory profiles of different opportunistic infections (OIs) among 125 immunocompromised patients admitted to a referral hospital in the eastern part of India. Different pathogens were isolated, identified and characterized using the laboratory gold standard methods. Oral candidiasis (88%) was found to be the most common OI, followed by tuberculosis (57%), enteropathogenic Vibrio (47%), cytomegalovirus infection (45%), cryptosporidial diarrhea (43%), Escherichia coli infection (42%) and other infections among the study subjects. Statistical analysis of the case studies shows 120/cumm median CD4+ blood cell count, and the OIs showed an inversely proportional occurrence to the CD4+ count of the immunocompromised patients. The spectrum and frequency of certain OIs highlight the urgency of studying HIV/AIDS in resource-limited countries where locally specific disease patterns may be observed. The purpose of the present investigation was the identification of such opportunistic pathogens, as we feel the HIV epidemic can be more effectively managed if physicians and health planners are aware of this information.

The purpose of the present study was to compare the incidence of childhood meningitis in regions with fewer than 2,000 inhabitants (rural regions) and regions with more than 2,000 inhabitants (urban regions) in the prefecture of Achaia in Western Greece during 1991-2005. Included were all 555 children hospitalized for meningitis. The criteria for bacterial meningitis were (i) positive blood/cerebralspinal fluid (CSF) culture, Gram stain, or latex agglutination and/or (ii) increased β-glucuronidase in CSF. In case of suspected bacterial meningitis, the following findings were considered: compatible clinical and laboratory findings, and whether or not a cure was achieved with antibiotic treatment and finally resulted in negative cultures. In cases of suspected viral meningitis, compatible clinical and laboratory findings were considered, together with observation of a cure without antibiotic treatment. Only 28 of 555 meningitis patients were from rural regions. The incidence per 10,000 children in rural and urban regions, respectively, was as follows: meningitis, 1.13 and 8.99; bacterial meningitis, 0.16 and 2.40; suspected bacterial meningitis, 0.52 and 3.00; and viral meningitis, 0.44 and 3.58. The incidence ratio for bacterial, suspected bacterial, and viral meningitis in urban versus rural regions was 14.85, 5.72, and 8.10, respectively. Only 2 of the 79 cases with a confirmed causative pathogen came from rural regions. In conclusion, compared to those living in urban regions, children living in rural regions are relatively spared from bacterial and viral meningitis.

We identified seven distinct subtypes of enterohemorrhagic Escherichia coli (EHEC) O157:H7 isolates that were derived from sporadic cases and outbreaks from multiple prefectures in Japan in 2005. A surveillance system utilizing pulsed-field gel electrophoresis (PFGE), PulseNet Japan, was used. Some strains showed indistinguishable PFGE patterns using another restriction enzyme (BlnI or SpeI) in each subtype of EHEC O157:H7 isolates that were routinely subtyped by the XbaI PFGE pattern. In order to examine the genotypic relatedness of these strains, we carried out a multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA). By using the MLVA system, we found that three of seven subtypes of EHEC O157:H7 strains that were isolated from sporadic cases dispersed across multiple prefectures within a few months showed indistinguishable PFGE patterns and identical MLVA types. Strains belonging to the other four subtypes of EHEC O157:H7 in the PFGE analysis were further classified into different clusters of EHEC O157:H7. Therefore, compared to PFGE, MLVA showed greater discriminatory power with respect to analysis of the isolates in this study.

With the introduction of varicella vaccination in India, surveillance of circulating varicella-zoster strains has gained significance. The aim of the present study was to achieve molecular characterization of circulating varicella-zoster virus (VZV) strains and differentiation from the Oka vaccine strain. In this study, the genotype of 100 clinical VZV strains was analyzed. Vesicle fluid was collected from patients with VZV infections (92 cases of varicella and 8 cases of herpes zoster). The PCR-RFLP analysis of two polymorphic loci–a PstI restriction site in ORF 38 and a BglI restriction site in ORF 54 was used to characterize and differentiate them from the vaccine strain. All the wild-type strains were positive for the PstI restriction site in ORF 38. This differentiated them from the Oka vaccine strain, which is PstI negative. The wild-type strains as well as the Oka vaccine strain were positive for the BglI restriction site in ORF 54. Thus, the genotype of all the VZV strains examined had the wild-type pattern represented as PstI⁺ BglI⁺. None of the strains had the PstI^- BglI⁺ genotype characteristic of the Oka strain or the PstI⁺ BglI^- wild-type pattern. To conclude, PstI and BglI serve as good reference markers in the genotyping of circulating varicella strains in India and serve to differentiate them from the vaccine strain as well as other wild-type strains.

The aim of the current study was to determine the possible crucial role of cancer antigen 125 (CA125) in the diagnosis of pulmonary tuberculosis (PTB). The CA125 levels of study and control groups were statistically compared. In a total of 146 patients that were included in the current study, 30 had active PTB, 37 inactive PTB, 28 community-acquired pneumonia (CAP), 25 pleural or pulmonary malignancies, and 13 patients exacerbation of chronic obstructive pulmonary disease. The mean CA125 levels in PTB, inactive PTB, CAP, and pleural-pulmonary malignancies were 118.46 ± 248.41, 40.80 ± 50.95, 47.76 ± 60.76, and 57.77 ± 65.59, respectively. For active-inactive discrimination of PTB, with a cut-off level of ≥35 U/ml, the sensitivity, specificity, positive predictive value, and negative predictive value of CA125 were 63, 59, 56, and 67%, respectively. Increased CA125 levels were detected in active PTB in the current results. The current results also show that high level CA125 should be reconsidered in the prediagnosis and/or discrimination of active and inactive PTB patients.

To evaluate the prevalence of Staphylococcus aureus carriage among healthy preschool children in our region, nasal swabs were collected from 5- to 7-year-old healthy children who were attending a day care center. Sociodemographic features and the data related with risk factors were obtained from the children's parents. Of the 200 children, S. aureus was isolated in 36 (18%) subjects. Methicillin-resistant S. aureus was isolated in 2 (5.6%) of these subjects, neither of whom had any identified risk factors. Antimicrobial susceptibility testing revealed that all tested strains were sensitive to gentamicin, vancomycin, trimethoprim-sulfamethoxazole, rifampicin, and mupirocin. Erythromycin, clindamycin, fusidic acid, and tetracycline resistance were determined to be 16.6, 8.3, 5.6, and 8.3%, respectively.

Hepatitis B virus (HBV) surface antigen mutations may lead to immune escape and eventually cause failure of immunization. In this report, we identified immune escape variants in immunized babies born to hepatitis B surface antigen (HBsAg) carrier mothers. A total of 68 babies were followed up for 2 years after the full course of vaccination; 2.9% (2/68) of babies were found to be infected with the variant HBV in spite of preexisting antibody to surface antigen (anti-HBs) at 24 months post immunization. Both infants were positive for HBV-DNA; sequencing results of the “a” determinant region of the surface gene revealed that both babies had point mutations at a different nucleotide position resulting in various amino acid substitutions. In addition, an intriguing variant having an addition-deletion mutation was observed in one of the babies. This is the first report to show the addition-deletion variant of HBV in India. However, the immunological significance of the above HBV variants needs to be further elucidated.

We report on a Mycobacterium monacense infection associated with a pulmonary tumor in a Chinese patient. To our knowledge, this is the first case of M. monacense described in a non-European patient with a tuberculosis-like disease. Further evaluation of the human pathogenic potential of M. monacense is needed.

Three groups of populations at high risk of human immunodeficiency virus (HIV) infection, i.e., injecting drug users (IDUs), truckers, and attendees of sexually transmitted disease (STD) clinic of Amritsar (Punjab), Northern India, were studied in order to determine the prevalence of HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV) infections. Of the 157 IDUs, 16.6, 17.8, and 33.7% were found to be positive for HIV, HBV, and HCV infections, respectively. HCV showed significant difference (P < 0.01) and very high rate (8.3%) of co-infection with HIV. In truckers, maximum seropositivity was associated with HIV (19%), i.e., significantly higher than that of HBV (6%, P < 0.01) and HCV (3%, P < 0.01). In STD clinic attendees, the highest rate of seroprevalence was that of HIV (4.3%), followed closely by that of HBV (3.7%), and HCV (2.6%). The present study emphasizes the need for early screening for HBV and HCV infections of all those also at high risk for HIV infection, and the results highlight the importance of relevant targeted interventions in these populations.

This study was conducted to determine the trends in Campylobacter antibiotic resistance occurring in our setting and to assess the differences in the isolates using patterns of plasmid profiles. One hundred Campylobacter jejuni strains of human and poultry origin isolated in 2002-2003 (phase A) and 2005-2006 (phase B) in the Kingdom of Bahrain were evaluated. Susceptibility to erythromycin, ciprofloxacin and tetracycline was determined, and plasmid extraction and polymerase chain reaction detection of the tet(O) gene was carried out. A single erythromycin-resistant isolate was identified, in sharp contrast to the high ciprofloxacin resistance which also showed an increment in phase B. Tetracycline resistance was higher in chicken (80.9%) compared to human (41.3%) isolates (P < 0.01). Most isolates harbored two plasmids (23 kb and 35 kb) with significant correlation between tetracycline resistance and plasmid carriage in chicken isolates. The findings show continued effectiveness of erythromycin for campylobacteriosis but an increasing trend of high ciprofloxacin and tetracycline resistance. Tetracycline resistance is most likely due to the transfer of plasmids carrying the tet(O) gene between isolates.

The purpose of this study was to determine the prevalence of multidrug-resistant Escherichia coli in clinical specimens. In addition, the existence of integrons in resistant isolates was assessed by amplification of intergase genes. Susceptibility of 200 isolates from five Shiraz hospitals and health centers to 13 antibiotics was determined by the Kirby-Bauer disk diffusion method. The majority of the bacteria were isolated from urine (70.5%) and stool (25.5%) specimens. Antibiotic resistance patterns were observed as follows: amoxicillin 63%, tetracycline 57.5%, co-trimoxazole 48%, cephalotin 40%, nalidixic acid 36%, ciprofloxacin 21%, nitrofurantoin 25%, norfloxacin 20.5%, gentamicin 18%, chloramphenicol 18%, ceftazidime 14%, amikacin 8.5% and imipenem 2%. Of 200 isolates tested, 165 (82.5%) were multidrug resistant. The frequency of multidrug resistance to more than 5 antibiotics was 24.2%. The existence of integrons was confirmed in 44.8% of isolates. Significant association between resistance to gentamicin, amikacin, cephalotin, nalidixic acid, ciprofloxacin, norfloxacin and co-trimoxazole with the existence of integrons was obtained by the PCR-RFLP method. These results showed that integrons may be partly responsible for multidrug resistance. Imipenem, amikacin and ceftazidime were the most effective antibiotics in vitro; however, the clinical efficacy of these antibiotics remains to be assessed.