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The aim of the present study was to describe the genetic diversity of Mycobacterium tuberculosis (M.tuberculosis) strains circulating in the region of Northern Greece. A total of thirty-seven M. tuberculosis clinical isolates were analysed by the spoligotyping method. According to the results, six clusters comprising seventeen strains were detected, and the remaining twenty strains showed unique patterns. The M.tuberculosis families according to SITVITWEB were distributed as follows: Haarlem (H) (27.0%); T (24.3%); Beijing (13.5%); Latin-America and Mediterranean (LAM) (5.4%) and S (2.7%). The remaining isolates (27%) did not match any isolates within the database and they were characterized as orphans. Regarding GenoType MTBDRplus results, two strains (5.4%) were Multi-Drug-Resistant, four strains (10.8%), were isoniazid monoresistant, while the remaining thirty-one strains (83.8%) were susceptible. In conclusion, in the region of Macedonia-Thrace (Northern Greece), there was high phylogenetic diversity among M. tuberculosis isolates. Molecular tools used and data presented can have regional and national impact on tuberculosis control.

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SETTING: Data on the relationship between pleural tuberculosis (TB) and anti-tuberculosis drug resistance are scarce. OBJECTIVE: To determine the patterns of drug resistance among pleural Mycobacterium tuberculosis isolates in Greece and the incidence of tuberculous pleural effusion (TPE) among patients with multidrug-resistant (MDR) or extensively drug-resistant (XDR) pulmonary TB. DESIGN: Drug susceptibility testing (DST) results recorded in the database of the National Reference Centre for Tuberculosis in Athens, Greece, over a 9-year period (2003-2011) were reviewed. Chest X-rays from hospitalised patients with pulmonary MDR/XDR-TB during the same period were also reviewed for the presence of TPE. RESULTS: Resistance to at least one first-line drug was observed in 11% of the cases (MDR-TB 3%, XDR-TB 1%), while 29% of the patients with pulmonary MDR/XDR-TB presented with TPE during the course of their disease, the majority ipsilateral to the lung lesions, which responded to guided anti-tuberculosis treatment. CONCLUSION: The prevalence of drug resistance among pleural M. tuberculosis isolates in Greece highlights the importance of DST prior to treatment selection in TPE patients. In our study population, TPE that developed in one third of the patients with pulmonary MDR/XDR-TB usually resolved with DST-guided anti-tuberculosis treatment.

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SETTING: Sotiria Chest Diseases Hospital (SCDH), a referral hospital in Athens, Greece, 2012. OBJECTIVE: To assess the completeness of the mandatory notification system for tuberculosis (TB) at the SCDH, and compare the observed and estimated annual incidence rates. DESIGN: Record linkage and the capture-recapture method were applied. Data sources were the registers from the national mandatory notification register (Hellenic Centre for Disease Control and Prevention [HCDCP]), the National Reference Laboratory for Mycobacteria (NRLM) and SCDH records. The log-linear model with the lowest Akaike information criterion was selected as the most valid statistical model. RESULTS: The observed and estimated TB under-reporting rates at the national level were respectively 55% (95%CI 49-60) and 75% (95%CI 71-78). The observed completeness of the HCDCP, NRLM and SCDH registers were respectively 45% (95%CI 40-51), 66% (95%CI 61-71) and 36.5% (95%CI 31-42). The estimated TB incidence rate was 15 cases per 100 000 (range 13-19/100 000), compared to the 4.9/100 000 rate officially notified. CONCLUSION: Adult TB incidence has been largely underestimated, and the TB burden is likely to be much higher than officially notified in our setting. A thorough review of the notification system should be carried out. The implementation of a network-based notification system and retraining of all relevant personnel is advised.

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BACKGROUND: A nosocomial outbreak in a 740-bed hospital in Athens, Greece, was investigated in January-February 2012. METHODS: Recommendations on infection control measures were given and two case-control studies were conducted among patients (study A) and health care workers (HCWs) (study B). Compliance to control measures was evaluated. RESULTS: The absence of a routine recording system of nosocomial-acquired gastroenteritis cases led to a 10 days delay in outbreak identification. In total, 63 gastroenteritis cases were identified; 30 HCWs and 33 patients. In the multivariable analysis of study A the disease incidence among patients was statistical significantly associated with a prior incident of vomitus in their room (OR=7.96, 95% CI=1.29-49.2). In study B, the incidence was associated with the history of direct contact with a symptomatic patient (OR=3.03, 95%CI 1.01-9.12). Twenty one (75%) of the symptomatic HCWs reported absence from work for a median of 2 days (range: 1-4). Seven (25.0%) continued to work despite being symptomatic. Only, 11.1% of patients were isolated or cohorted after developing symptoms. In-hospital virological testing was not feasible and one specimen sent to a university laboratory was positive for norovirus. CONCLUSIONS: An appropriately designed protocol regarding the detection, the management and the laboratory investigation of nosocomial gastroenteritis outbreaks should be followed in order effective containment to be reassured. Hippokratia 2014; 18 (3): 204-208.

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We present the first fatal case of extensively drug-resistant tuberculosis (XDR-TB) in an injecting drug user (IDU) in Athens, Greece, co-infected with human immunodeficiency virus and hepatitis C virus and discuss the implications for public health. Despite immediate initiation of treatment, the patient's condition gradually deteriorated and he died 16 days after hospital admission because of multiple organ failure. The contact tracing investigation revealed no further infections among the patient's contacts.

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BACKGROUND: Cystatin C is a marker of renal function that appears to be associated with inflammation. The aim of the present study was to investigate whether there is any relationship between cystatin C, total and differential leukocyte count and other inflammatory markers. METHODS: Cystatin C, creatinine, high sensitivity C-reactive protein (hs-CRP), haptoglobin, ferritin, serum albumin, glucose, total cholesterol, HDL and triglycerides together with total and differential leukocyte count were determined in 490 adults (46 ± 16 years, 40% men) who underwent a typical health examination. Glomerular filtration rate was estimated by the simplified Modification of Diet in Renal Disease formula. Anthropometric and lifestyle characteristics were also recorded. RESULTS: After adjustment for demographic risk factors, comorbid health conditions and renal function, a positive and independent relationship of serum cystatin C levels with peripheral monocyte blood count (regression coefficient ± SE: 12 ± 3.38, P < 0.001) and white blood count (0.616 ± 0.278, P= 0.027) was evident. In this multiple linear regression analysis, other inflammatory markers (i.e. hs-CRP, haptoglobin, ferritin, albumin) did not seem to affect cystatin C blood levels. CONCLUSION: The results of this study demonstrated that monocytes, which play an important role in chronic inflammation and atherosclerosis, were independently related with cystatin C concentrations. This finding may provide a plausible link for the usefulness of cystatin C in predicting increased cardiovascular risk.

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BACKGROUND: The role of Helicobacter pylori infection and especially of the cytotoxin-associated gene A (CagA) product strain in peptic ulcer bleeding among non-steroidal anti-inflammatory drugs (NSAIDs) users remains controversial. METHODS: A case-control study was carried out including 191 consecutive chronic NSAIDs users admitted to hospital because of peptic ulcer bleeding. Peptic ulcer was verified by endoscopy. Controls comprised 196 chronic NSAIDs users without signs of bleeding of similar age and gender to cases. Multivariate regression analysis was performed for further evaluation of the relationship between H. pylori, CagA status and other risk factors. RESULTS: H. pylori infection was present in 121 (63.4%) cases compared with 119 (60.7%) controls (odds ratio (OR) = 1.14, 95% CI, 0.76-1.72). CagA-positive strains were found to be significantly more frequent in cases than in controls (65/106 versus 41/99 P = 0.008). Current smoking (OR = 2.65; 95% CI, 1.14-6.15; P= 0.02), CagA status (OR = 2.28; 95% CI, 1.24-4.19; P = 0.008), dyspepsia (OR = 6.89; 95% CI, 1.84-25.76; P = 0.004) and past history of peptic ulcer disease (OR=3.15; 95% CI, 1.43-6.92; P=0.004) were associated significantly with increased risk of bleeding peptic ulcer. CONCLUSIONS: The results suggest that CagA-positive H. pylori infection is associated with a more than 2-fold increased risk of bleeding peptic ulcer among chronic NSAIDs users.

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Mycobacterium gordonae was isolated as a light growth from bronchoalveolar aspirates from nine patients over 12 months. All patients were in one hospital, and had been bronchoscoped for suspected malignancy. None of the patients had symptoms or radiographic findings of mycobacterial infection. The isolates were characterized by biochemical tests and molecular hybridization. Random amplified polymorphic DNA analysis (RAPD) was used to test whether the strains had a common origin. All the isolates generated four to eight fragments, and almost all presented distinct RAPD patterns. Antimicrobial resistance patterns to six agents confirmed that the isolates were unrelated. Thus epidemiologically unrelated strains of M. gordonae can exist as contaminants in the same department over a relatively short time frame. RAPD analysis is easy to perform, gives rapid results, and can be used for epidemiological analysis of M. gordonae isolates.

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Rhodococcus equi is a facultative intracellular bacterium that can cause pneumonia in both young horses and immunocompromised humans. In this study, we have tried to determine the T-cell populations that recognize this pathogen during murine infection, as well as the bacterial antigens recognized by these cells. When BALB/c mice were hyperimmunized with a virulent R. equi strain, we did not observe preferential expansion of a particular T-cell subset in their spleens. However, when the splenic T lymphocytes of the hyperimmunized BALB/c mice were cultured in the presence of killed bacteria, we found that alpha/beta CD4+ T cells proliferated and exhibited increased levels of the interleukin-2 receptor (IL2R). In order to ensure antigen specificity, two different controls were included in these experiments: (i) T-cell proliferation and expression of the IL2R in the presence of the major membrane constituent of Bacillus megaterium were studied comparatively with the presence of the R. equi bacterial antigen, and (ii) T-cell proliferation and expression of the IL2R from naive, non-infected mice in the presence of bacterial antigens were compared to those observed in hyperimmunized mice. In our study, the T cells from hyperimmunized mice did not significantly proliferate nor were they activated in the presence of non-related bacterial antigens, and T cells from naive mice were not found to significantly recognize R. equi antigens. When we studied the localization of R. equi antigens that could stimulate the in vitro proliferation and activation of T cells, we found that they were constituents of the bacterial cell wall and the cytoplasm, but they were not excreted in the culture medium. For these experiments, T-cell recognition of the bacterial antigens in hyperimmunized mice was compared to that of naive mice. With T-cell immunoblotting, we found that T-cell proliferation and activation were obtained with proteins having molecular masses of approximately 65, 43, 30, 22-27 and 15-17 kDa. It is noteworthy that among the recognized bacterial antigens, some have been described as being associated with virulence.

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