Efi Petinaki Associate Progessor MIcrobiology Section, Medical School, University of Thessalia E-mail: petinaki@med.uth.gr; petinaki@hotmail.com
Research Interests: Microbiology, Molecular Biology, Epidemiology, Human Infections and Clinical Immunology.
Member of Research Projects co-funded by the European Social Fund - National Resources - Seventh Framework Programme of the European Community for Research. Author of chapters in Books. Supervisor in 9 PhD. Author in 129 International and 82 Hellenic Conferences.
Author of 68 papers in Intrernational Journals with I.F. Number of Citations : 551 (Scopus, ISI)
Abstract: Loop-mediated isothermal amplification (LAMP) is a recently developed molecular method that has been successfully implemented in the detection of Mycobacterium tuberculosis in clinical specimens. LAMP has several advantages, such as rapidity, high sensitivity, ease of application and cost-effectiveness. As a result, it is anticipated that its use for the detection of M. tuberculosis is likely to become widespread, especially in low-resource countries. The present review aimed to present this method and all of the available information on its implementation in the detection of M. tuberculosis in clinical specimens.
Abstract: Abstract Female genital tuberculosis is an uncommon type of tuberculosis that can lead to infertility. The present review describes the disease, reports available epidemiological data, and focuses on examinations and procedures necessary for the early diagnosis and the management of this curable disease.
Abstract: Tonsillar disease (recurrent tonsillitis and/or tonsillar hypertrophy) is one of the most common human disorders, with Streptococcus pyogenes (group A beta-hemolytic streptococcus [GAS]) and Haemophilus influenzae representing the most common pathogens. Until now, no study has investigated why some individuals are more susceptible to tonsillar infections caused by specific bacteria than others. The aim of this study was to uncover possible associations between common Toll-like receptor gene (TLR) polymorphisms and tonsillar disease. The TLR2-R753Q, TLR4-D299G, and TLR4-T399I polymorphisms were determined in a cohort of 327 patients subjected to tonsillectomy due to recurrent tonsillitis (n = 245) and tonsillar hypertrophy (n = 82) and 245 healthy bone marrow donors. Associations of the aforementioned polymorphisms with the isolated bacterial strains after tonsillectomy were also investigated. Interestingly, carriers of the TLR4 polymorphisms displayed an approximately 3-fold increased risk for GAS infections (for TLR4-D299G, odds ratio [OR] = 2.81, 95% confidence interval [CI] = 1.16 to 6.79, P = 0.038; for TLR4-T399I, OR = 3.01, 95% CI = 1.29 to 7.02, P = 0.023), and this association was more profound in patients with recurrent tonsillitis. On the contrary, the presence of the TLR4-T399I polymorphism was associated with a 2-fold decreased risk of Haemophilus influenzae carriage (OR = 0.38, 95% CI = 0.15 to 0.96, P = 0.038). In the end, no significant differences were observed, considering the genotype and allele frequencies of the above-mentioned polymorphisms, between patients and controls. Our findings indicate that, regarding tonsillar infections, TLR4 polymorphisms predispose individuals to GAS infection, while they are protective against Haemophilus influenzae infection. This result further elucidates the role that host immune genetic variations might play in the susceptibility to common infections and tonsillar disease.
Abstract: Multidrug-resistant Acinetobacter baumannii (MDR-AB) infections are difficult to treat owing to the extremely limited armamentarium. The present review reports all available treatment options against MDR-AB, including single molecules, combination schemes, and alternative modes of antimicrobial administration. Additionally, a group of recently reported peptides with anti-MDR-AB activity is described.
Abstract: Polyphasic identification combined with antifungal susceptibility study of Aspergillus spp. clinical isolates comprising 102 common and rare members of Sections Fumigati, Flavi, Terrei, Nidulantes, Nigri, Circumdati, Versicolores and Usti is the first in Greece. High amphotericin B MIC (>2μg/ml) exhibited 17.6% of strains. Itraconazole, posaconazole and voriconazole MIC>4μg/ml demonstrated 1%, 5% and 0% of the isolates respectively. Anidulafungin, micafungin and caspofungin MECs≥2 μg/ml were correspondingly recorded for 4%, 9% and 33% of the strains.
Abstract: Because of their frequent isolation in the routine laboratory and their increasing clinical significance, fast and accurate species identification of staphylococci may be required; this can only be achieved by automated systems. A total of 147 clinical isolates (52Â Staphylococcus aureus, 50 Staphylococcus epidermidis, and 45 other coagulase-negative staphylococci [CoNS]) were first identified by molecular methodology and then comparatively tested by Vitek 2 (new colorimetric identification card) and Phoenix using the novel 0.25Â McFarland and the standard 0.50 McFarland inoculum protocols. All S. aureus isolates were accurately identified. Vitek 2 identified correctly all S. epidermidis and 93.3% of the other CoNS, whereas the respective rates were 86% and 82.2% for Phoenix's standard and 92% and 82.2% for the novel protocol. It appears that both systems provide excellent identification of S. aureus, but Vitek 2 recognizes CoNS species more accurately than Phoenix. The 0.25 McFarland protocol does not improve system performance.
Abstract: A total of 359 vancomycin-resistant enterococci (344 Enterococcus faecium and 15 E. faecalis) collected during 2007 from eight tertiary-care hospitals in Greece were analysed for genotypic characteristics. Four common clones, ST412, ST203, ST16 and ST17, were identified among E. faecium and one clone, ST28, among E. faecalis strains.
Abstract: Mycobacterium arupense is a novel mycobacterium species. It was first identified from clinical specimens in 2006 and since then there have been only two reports of its recovery from clinical samples. In the present case M. arupense was isolated from the sputum of a 62-year-old man with a malignant mass in his left kidney, who presented with a one-month history of recurrent fever, dyspnea and haemoptysis. M. arupense was identified with sequencing of hsp65 and 16S rRNA genes. In the present study, its biochemical profile along with its resistance status and hsp65 RFLP analysis is presented.
Abstract: Staphylococcus lugdunensis is an unusually virulent coagulase-negative species, associated with severe infections. The present report describes the development of a single-step, species-specific PCR protocol for S. lugdunensis identification. fbl gene, encoding a fibrinogen-binding adhesin, was exploited and assessed as a suitable nucleic acid target. The gene was detected in all 17 S. lugdunensis isolates examined, while no amplification product was obtained from 98 isolates representing 11 staphylococcal and 17 nonstaphylococcal species. Forty-seven percent of the S. lugdunensis strains produced a positive slide coagulase reaction, which is consistent with varying levels of Fbl protein expression within the species.
Abstract: Prosthetic valve and pacemaker lead endocarditis by Staphylococcus lugdunensis remain very rare, while the former is associated with an ominous prognosis. Two cases involving a prosthetic aortic valve and a pacemaker lead, respectively, are reported. Despite disease severity and delayed diagnosis, patients recovered fully with combined antimicrobial and surgical treatment.
Abstract: Mycobacterium thermoresistibile is a non-tuberculous mycobacterium strongly associated with human infections. Since 1966, there have only been six reports of its isolation from clinical samples. We report on the first case from Europe and review all the previous cases. Identification was achieved with sequencing of the 16S rRNA and hsp65 genes. This study presents its phenotypic and biochemical profile, susceptibilities to selected antibiotics and hsp65 polymerase chain reaction-restriction fragment length polymorphism profile with BsteII and Hae III .
Abstract: Three molecular assays were evaluated for the direct detection of Mycobacterium tuberculosis complex bacteria in 125 respiratory and 22 nonrespiratory samples. The overall sensitivities obtained were as follows: GenoType MTBDRplus, 97.9%; GenoType Mycobacteria Direct, 93.7%; Gen-Probe Mycobacterium tuberculosis Amplified Direct Test, 89.6%. The specificity of the assays used was 100%.
Abstract: Ralstonia pickettii is an infrequent pathogen of invasive infections in healthy individuals. The microorganism is supposed to be of relatively low virulence, but can cause infections, mainly of the respiratory tract, in immunocompromised and cystic fibrosis patients. Ralstonia pickettii has also been associated with hospital outbreaks related to contamination of products used for medical care and laboratory diagnosis.
Abstract: Visceral leishmaniasis (VL) remains a public health problem in most countries bordering the Mediterranean basin. Its diagnosis is challenging and often delayed, as the main clinical picture is often indistinguishable from that of other infectious and non-infectious diseases. Herein, we report two unusual cases of VL that presented with several characteristics of autoimmune hepatitis (AIH). Neither patient had a history of fever, only generalized symptoms accompanied by polyclonal hypergammaglobulinemia, cytopenias, signs of portal hypertension, elevated transaminases, and high titers of antinuclear and smooth-muscle autoantibodies (SMA) with reactivity against filamentous actin (F-actin), which has been recognized as specific to AIH. A clinical diagnosis of AIH was considered, but a bone marrow biopsy was performed before a liver biopsy to exclude a primary bone marrow disease. The biopsy led to the diagnosis of VL. The diagnosis was further confirmed by IgG antibodies against Leishmania spp. using ELISA and PCR-based assays. Treatment with amphotericin in the first case and pentamidine in the second (because of a severe reaction to amphotericin) was effective. From the clinical point of view, it should be emphasized that, in cases with high titers of anti-F-actin AIH-specific SMA accompanied by polyclonal hypergammaglobulinemia, the possibility of AIH should be cautiously differentiated from VL; this distinction is of paramount importance because initiation of immunosuppression for AIH treatment would be detrimental to a patient with underlying leishmaniasis. Therefore, in such cases and in areas where the disease is still present, it seems rational to exclude VL before starting any immunosuppressive therapy.
Abstract: The autograft preparation process for anterior cruciate ligament reconstruction has a potential for graft contamination. The purpose of this study was to evaluate the possibility of contamination of the bone-patellar tendon-bone and hamstring tendon autograft during preparation for anterior cruciate ligament reconstruction.
Abstract: Clonal analysis and PCR screening for the presence of Panton-Valentine leukocidin (PVL) genes was performed among 694 methicillin-resistant Staphylococcus aureus (MRSA) cases collected during a 2-y period in Greece. The detection rate of PVL-positive MRSA is high, both in the community and in hospital. Clonal analysis revealed the predominance among the PVL-positive strains of the clonal complex CC80 (ST80-IV) and the emergence of ST377 clone carrying agr1 allele and SCCmec type V.
Abstract: Although QuantiFERON-TB Gold (QFT-G) has been approved for the diagnosis of latent tuberculosis infection (LTBI), there are limited data regarding its performance in routine clinical practice. The aim of this study was to compare QFT-G 'In Tube' results, based on stimulation with Mycobacterium tuberculosis-specific antigens, with tuberculin skin test (TST) results in an unselected hospital-based Greek population.
Abstract: Coagulase-negative staphylococci (CoNS) are frequently encountered pathogens in hospital environment. Dialysis patients, often carrying central venous catheters, are prone to CoNS infections. Methicillin-resistant (MR) staphylococci in hospitals are resistant to multiple antibiotics and may cause an overall increase in the incidence of staphylococcal infections rather than simply replacing the more susceptible strains. The aim of this study was to evaluate the antimicrobial resistance and the clonal relatedness of all clinically significant CoNS isolates recovered from haemodialysis patient infections treated in a tertiary care centre, the University Hospital of Larissa, in central Greece. In addition, the CoNS isolates from carriers among health-care workers of the local haemodialysis unit were tested.
Abstract: The aim of this study is to establish a new experimental model of hematogenous implant-related infection (IRI) by a community-acquired methicillin-resistant S. aureus (CA-MRSA) strain. Cylindrical porous tantalum intramedullary implants were inserted in the proximal right tibia of 30 male white rabbits after administration of antibiotic prophylaxis. Four weeks later and without antibiotic prophylaxis, 20 animals received 1 ml of inoculum of two different concentrations (study groups A and B) of CA-MRSA strain through an ipsilatelar femoral artery catheter. The remaining 10 received normal saline instead (control group C). Surviving animals were sacrificed 4 weeks later. Sterile bone, bone marrow biopsies, and implants were harvested for culture and histological evaluation. Ten animals receiving 5 x 10(8)cfu/ml (group A) died within 48-72 h due to septic shock. Blood cultures were positive; histology demonstrated acute infection. Ten animals received bacterial load of 3 x 10(8)cfu/ml (group B) and all survived; two had negative Gram-stain and cultures but PCR and RT-PCR results demonstrated the viability of the microorganisms, while periprosthetic osteolysis and histological evaluation indicated subacute osteomyelitis; eight animals established periprosthetic infection, osteomyelitis, and septic arthritis documented by positive Gram-stain, cultures, subperiosteal reaction, and chronic infection on histology. Control group specimens demonstrated no signs of infection. Histopathological semiquantitative scoring was used to compare the three groups. Comparison of groups A and B with control group and between group A and B showed statistically significant difference (p < 0.05) in all parameters except for periosteal reaction between groups B and C (p = 0.354). This novel, reproducible experimental model will facilitate the study of hematogenous CA-MRSA IRIs.
Abstract: During 2005-2006, a total of 865 Enterococcus faecium isolated from patients from eight Greek hospitals were tested for susceptibility to quinupristin/dalfopristin (Q/D). Among them, 250 genetically unrelated strains (28.9%) were found to be intermediate-resistant to Q/D (minimum inhibitory concentration (MIC) 1.5-4 mg/L); all were resistant to dalfopristin (MIC=16-64 mg/L), whilst 69% were resistant to quinupristin, carrying the ermB gene. No strain was found to carry any of the known genes, such as vatE and vatD, involved in Q/D resistance, indicating that a non-transferable undetermined mechanism is responsible for the expression of low-level Q/D resistance. The high percentage of Q/D-intermediate-resistant E. faecium in Greece was not associated with prior consumption of the agent or with the veterinary use of virginiamycin.
Abstract: Streptococcus uberis UCN 42, isolated from a case of bovine mastitis, was intermediately resistant to lincomycin (MIC = 2 microg/ml) while remaining susceptible to clindamycin (MIC = 0.06 microg/ml) and erythromycin. A 1.1-kb SacI fragment was cloned from S. uberis UCN 42 total DNA on plasmid pUC 18 and introduced into Escherichia coli AG100A, where it conferred resistance to both clindamycin and lincomycin. The sequence analysis of the fragment showed the presence of a new gene, named lnu(D), that encoded a 164-amino-acid protein with 53% identity with Lnu(C) previously reported to occur in Streptococcus agalactiae. Crude lysates of E. coli AG100A containing the cloned lnu(D) gene inactivated lincomycin and clindamycin in the presence of ATP and MgCl(2). Mass spectrometry experiments demonstrated that the lnu(D) enzyme catalyzed adenylylation of clindamycin. A domain conserved in deduced sequences of lincosamide O-nucleotidyltransferases Lnu(A), Lnu(C), LinA(N2), and Lin(D) and in the aminoglycoside nucleotidyltransferase ANT(2'') was identified.
Abstract: All Staphylococcus aureus isolated during 2003-2006 at a university hospital in Thessalia, central Greece, from ulcerative upper-extremity infections were tested for the presence of PVL gene and for possible clonal relationship to validate the role of PVL toxin in the clinical features of these infections and also to establish preventive measures towards limiting the spread of such strains among close contacts.
Abstract: Recurrent tonsillitis is 1 of the common human infectious diseases worldwide, but, to date, its pathogenesis remains unclear. Although Streptococcus pyogenes (GAS) is involved in recurrent bouts of acute tonsillitis, conventional cultures usually fail to isolate it. The purpose of this study was to clarify whether the deep tonsillar tissues of patients with recurrent tonsillitis might harbour GAS, resulting in reinfections. Deep tonsillar tissues obtained from 285 patients with recurrent tonsillitis and 172 patients with tonsillar hypertrophy, who had undergone tonsillectomy, were examined for the presence of GAS, using conventional and molecular methods. Cultures from all patients were negative for GAS. GAS DNA was found in the deep tonsillar tissues of 57 out of 285 patients with recurrences (20%), and GAS RNA, indicating the viability of GAS, was detected in 47 of them (82%). On the other hand, Haemophilus influenzae DNA was found in 15% and 16% of patients with recurrences and hypertrophy, respectively; but no Haemophilus influenzae RNA presence was detected. The low level of presence of GAS in patients with recurrent tonsillitis indicates that other unknown factors may be responsible for the recurrences.
Abstract: A total of 10420 Gram-positive cocci (including staphylococci, enterococci and various groups of streptococci) collected from clinically significant specimens in ten Greek hospitals during 2006--2007 were tested for their susceptibility to daptomycin. The minimum inhibitory concentration (MIC) was determined by the broth microdilution method. Daptomycin demonstrated very high activity against Enterococcus faecalis (MIC at which 50% of the isolates were inhibited (MIC50) = 1mg/L and MIC at which 90% of the isolates were inhibited (MIC90) = 1.36 mg/L), Enterococcus faecium (MIC50 = 1.36 mg/L and MIC90 = 1.90 mg/L), Streptococcus pyogenes (MIC50 = 0.12 mg/L and MIC90 = 0.50mg/L), Streptococcus agalactiae (MIC50 = 0.09 mg/L and MIC90 = 0.12 mg/L), Streptococcus pneumoniae (MIC50 = 0.24 mg/L and MIC90 = 0.5 mg/L) and viridans group streptococci (MIC50 = 0.50 mg/L and MIC90 = 0.89 mg/L). Resistance to linezolid and vancomycin for enterococci and to penicillin for streptococci appears to be independent of reduced susceptibility to daptomycin. On the other hand, daptomycin was also active against meticillin-resistant Staphylococcus aureus (MIC50 = 0.44 mg/L and MIC90 = 0.78 mg/L) and meticillin-resistant coagulase-negative staphylococci (MIC50 = 0.24 mg/L and MIC90 = 0.44 mg/L); however, 0.9% of the staphylococci tested had an MIC > 1mg/L, which is the Clinical and Laboratory Standards Institute breakpoint proposed for susceptibility. For all tested organism groups, resistance to daptomycin was not associated with glycopeptide resistance.
Abstract: We report the case of a 56-y-old male admitted with a left-sided post-pneumonic empyema. Clostridium sordellii DNA was directly detected in its pleural fluid by a broad-range 16S rRNA PCR, after 24 h of specimen collection. This is the third case of pleural infection caused by C. sordellii in the literature.
Abstract: Three collections of Staphylococcus aureus isolates (n = 1,058) were investigated to assess the spread of Panton-Valentine leukocidin (PVL)-producing strains in Greece and their association with skin and soft-tissue infections (SSTIs). The isolates were collected during 2001-2003 from inpatients and outpatients with invasive infections in two distinct geographical areas. Clonal types were identified according to their ClaI-mecA::ClaI-Tn554::pulsed-field gel electrophoresis patterns, and the presence of the lukS-PV and lukF-PV genes was assessed by PCR. In total, 287 (27%) S. aureus isolates carried the PVL genes: 45% of methicillin-resistant S. aureus (MRSA) and 12% of methicillin-sensitive S. aureus (MSSA). All the PVL-positive MRSA isolates belonged to a single clone that was disseminated in the community and hospitals. The PVL-positive MSSA isolates were polyclonal, with 14 of 65 isolates being associated with hospital-acquired infections. The community-acquired isolates were from SSTIs, while the hospital-acquired isolates were associated with surgical wound infections, especially those involving prosthetic devices. Thus, a unique clone of PVL-positive MRSA has spread in both the community and the hospital setting in Greece, and has replaced older clonal types.
Abstract: Idiopathic pulmonary fibrosis (IPF) is a fibroproliferative disorder. Cytokines contribute an important but yet undefined role to its pathogenesis.
Abstract: Bacterial DNA was detected directly in the serum of a patient with endocarditis by broad-range 16S rRNA PCR followed by sequencing and analysis of the results by the BLAST search. Using these methods, Cardiobacterium hominis was identified in 2 days from the date of serum collection. The microorganism was also isolated and identified using conventional methods (bacterial culture and biochemical tests) 17 days from the date of sample collection. This is the first report showing the direct detection of C. hominis in a patient's serum using molecular-based methods, emphasizing their potential usefulness as additional and rapid diagnostic tools for the detection and identification of fastidious bacteria.
Abstract: We describe a case of necrotizing pneumonia in Greece caused by a community-acquired methicillin-resistant Staphylococcus aureus (MRSA) producing Panton-Valentine leukocidin (PVL) carrying the staphylococcal chromosome cassette mec (SCCmec) type V. The pulse field gel electrophoresis (PFGE) pattern of this strain differed significantly from the PVL-positive multi-locus sequence typing (MLST)-80 clone of MRSA, which predominates in Greece and in Europe. Further analysis of this strain revealed that it belonged to the agr1 allele type, and its resistance to tobramycin, gentamicin, and kanamycin was associated to the presence of aac-(6')-Ie+aph(2') gene. This is the 1st report that documents the emergence of CA-MRSA with PVL genes in combination with the SCCmec type V in Europe.
Abstract: Methicillin-resistant coagulase-negative staphylococci (MR-CNS) (n = 132), isolated from pre-term neonates, were analysed to determine their antibiotic resistance patterns, clonal distribution, biofilm production and the presence of the ica operon. All MR-CNS were multiresistant, and 89% produced slime. A major clone was identified (77 isolates) among 115 Staphylococcus epidermidis isolates. Ten of 16 Staphylococcus haemolyticus isolates also belonged to a single clone. Most (80%) slime-positive isolates possessed all the ica genes tested, while the remaining 23 (20%) had a variety of gene combinations. The entire ica cluster was detected in three of 15 slime-negative isolates. One major and two minor slime-positive, multiresistant MR-CNS clones had disseminated among hospitalised pre-term neonates.
Abstract: Two commercial methods for the identification of coagulase-negative staphylococci (CNS) were compared with the restriction fragment length polymorphism (RFLP) of the amplified tuf gene, which served as the reference method.
Abstract: Risk factors for atherosclerosis such as hypertension, type 2 diabetes, obesity and dyslipidemia affect endothelial function and stimulate adhesion molecules expression. The aim of the study was to examine endothelial activation in type 2 diabetes and hypertension as indicated by adhesion molecule levels and further to investigate whether the coexistence of the above conditions has a different effect.
Abstract: Psoriasis is a chronic debilitating cutaneous disorder that affects both sexes and appears clinically as inflamed, edematous skin lesions covered with a silvery white scale. Strong evidence suggests that immune mechanisms are implicated in its pathogenesis, such as persistent activation of T-lymphocytes, excessive proliferation of keratinocytes and reactivation of proto-oncogenes and other elements. Additionally, several recent studies have demonstrated that cytokines play a significant role in the pathogenesis of the disease, as they can be found in the affected skin of psoriatic patients. In this study we evaluated levels of circulating cytokines in the serum of 45 Greek psoriatic patients before initiation of treatment and compared the results with those in 45 healthy volunteers. According to our findings interleukin (IL)-2, IL-10, IL-12 and tumor necrosis factor-alpha (TNF-alpha) levels were statistically significantly elevated in the serum of psoriatic patients before therapy compared with those of controls. IL-6 serum levels did not differ between psoriatic patients and healthy volunteers. Conversely, interferon-gammaserum levels of psoriatic patients were statistically significantly lower than those of healthy volunteers.
Abstract: Macrolide, lincosamide and streptogramin type B (MLS(B)) resistance was determined in Staphylococcus aureus clinical isolates from two University Hospitals.
Abstract: Background: Experimental studies demonstrate that hepatitis B virus may induce nitric oxide (NO) production in infected hepatocytes. Its presence in acute hepatitis B patients has not been studied. Methods: Serum levels of nitric oxide and its regulatory pro-inflammatory cytokines were detected in 15 patients with uncomplicated acute hepatitis B, 19 blood donors and 15 chronic hepatitis B patients. Cytokines were determined with an immunoassay. Nitric oxide was measured as the serum metabolic products of nitrates and nitrites using a modification of the Griess reaction. Results: All detected cytokines were increased in acute hepatitis B patients compared to healthy controls (p<0.001 for TNF-alpha, p<0.05 for IL-6, p<0.001 for IL1-beta and p<0.001 for IFN-gamma). High serum levels of nitric oxide were found in acute hepatitis B patients (156.96+/-9.76 micromol/l) compared to healthy controls (51+/-6.2 micromol/l, p<0.001) and chronic hepatitis B patients (63.97+/-3.78 micromol/l, p<0.001). No significant correlations were found between NO, cytokine levels and transaminases. Conclusions: High levels of nitric oxide and its regulatory cytokines were found in a group of patients with uncomplicated acute hepatitis B. The exact role of NO in the disease pathogenesis and outcome needs to be studied further.
Abstract: The accuracy of the Bactec MGIT 960 system for susceptibility testing of 177 clinical isolates of Mycobacterium tuberculosis to first line drugs (isoniazid, rifampicin, ethambutol and streptomycin) was compared with the agar reference method. The sensitivity, the ability to detect resistance, of the MGIT system was 100%, while the specificity, the ability to detect susceptibility, ranged from 98.6% to 100% for all drugs tested.
Abstract: Abnormal endothelial function is well known in patients with type 2 diabetes mellitus and thought to induce macroangiopathy. Increased levels of adhesion molecules have been found in type 2 diabetic patients and it has been suggested that they play an important role in the initiation of atherosclerosis. The aim of the present study was to clarify the relationship between objectively proven peripheral arterial disease (pAVD) and serum levels of soluble adhesion molecules in patients with type 2 diabetes mellitus.
Abstract: In order to test the serum levels of ICAM-1, ICAM-2 and ICAM-3 in patients with idiopathic pulmonary fibrosis (IPF), twenty patients with IPF and eleven with secondary interstitial fibrosis (SIF), as well as forty healthy volunteers (HV) were studied. Serum intracellular adhesion molecules (ICAM) 1, 2 and 3 were assessed by ELISA. Functional respiratory tests, which included spirometry and lung diffusing capacity were simultaneously performed. Median values of serum ICAM-1 and ICAM-2 were higher in the patients' than in the healthy volunteers' (HV) group: IPF group: 946.60 ng/ml and 400.14 ng/ml; SIF group: 901.58 ng/ml and 378.27 ng/ml; HV group: 308.40 ng/ml and 217.55 ng/ml, respectively (p<0.05). ICAM-3 serum levels were equal between the three groups. ICAM-2 negatively correlated to DLCO values. (p<0.005). It can be concluded that ICAM 1 and 2 are elevated in the sera of patients with pulmonary fibrosis. ICAM-2 might be associated with a more impaired clinical status.
Abstract: Raised serum levels of adhesion molecules are believed to reflect endothelial activation and may contribute to the development of diabetic vascular complications. The aim of this study was to clarify the association between soluble adhesion molecules levels and retinopathy in type 2 diabetic patients. Levels of soluble E-selectin, ICAM-1 and VCAM-1 were measured by enzyme-linked immunosorbent assay (ELISA) in 47 type 2 diabetic patients classified in two subgroups according to the presence (n=34) or absence (n=13) of retinopathy as determined by fundus ophthalmoscopy; 22 control subjects were also studied. Soluble E-selectin levels were significantly elevated in both diabetic subgroups compared to control subjects (p<0.01), while no significant difference was found in sICAM-1 and sVCAM-1 levels. However, sE-selectin, sICAM-1 and sVCAM-1 levels were comparable in diabetic subgroups. The progression of retinopathy was not associated with an increase in soluble adhesion molecules levels. Stepwise multiple regression analysis revealed that only diabetes duration and microalbuminuria were independent determinants of retinopathy (p<0.01). Our results confirm the contribution of endothelial activation in the development of diabetic complications as indicated by increased levels of soluble adhesion molecules. However, a direct implication of adhesion molecules in the pathogenesis or progression of type 2 diabetic retinopathy cannot be supported.
Abstract: Streptococcus agalactiae typically induces serious infections in pregnant women and newborns. Nonpregnant adult patients can also be infected and mortality rate exceeds 40%. CAPD peritonitis is very rarely induced by S. agalactiae. Seven cases have been described previously and all had a very severe course, which included bacteremia, septic shock and death. A 27-year-old male with end-stage renal disease due to membranoprolipherative glomerulonephritis type I, who was on CAPD for 17 months, was admitted with the clinical and laboratory picture of CAPD peritonitis. Severe abdominal pain, shaking chills and fever 38.5 microC were also observed at presentation. Streptococcus agalactiae was isolated from the peritoneal fluid and blood culture was sterile. Under treatment with ceftazidime and tobramycin (i.p.) and vancomycin (i.v.) cultures became negative after 48 hours, abdominal symptoms resolved after 12 days and WBC count in the dialysate normalized after 14 days. As a possible source of infection the patient's partner was shown to be a vaginal carrier of a clone of S. agalactiae identical to that isolated in the peritoneal fluid. S. agalactiae is a rare cause of CAPD peritonitis with potentially very serious consequences. Anal or genital tract colonization is, in general, the source of contamination with S. agalactiae. The microbiological findings in the case presented here suggest that colonization of the patient or of his close environment may be important in the pathogenesis of S. agalactiae-induced CAPD peritonitis.
Abstract: The possible contribution of metallo-beta-lactamases in the frequent detection of carbapenem-resistant Pseudomonas aeruginosa isolates in a tertiary Greek hospital in Central Greece was investigated.Materials and methods: All carbapenem-resistant (imipenem- and/or meropenem-resistant) P. aeruginosa isolates recovered from separate patients during a 1 year period in the Clinical Microbiology Laboratory at the University Hospital of Thessaly, Larissa, Greece, were studied for metallo-beta-lactamases. They were tested by Etest MBL, PCR analysis and nucleotide sequencing. DNA fingerprints were obtained by pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA.
Abstract: Initiation of a fibrotic process has been suggested as part of the intestinal response to chronic inflammation in inflammatory bowel disease. YKL-40 has been proposed as a new serum marker of fibrosis. We studied compared the serum levels of YKL-40 in patients with ulcerative colitis or Crohn's disease with inflammatory and healthy controls.
Abstract: The combined use of Bactec MGIT 960 system and a PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis was assessed for the rapid detection and identification of mycobacteria from clinical samples. The diagnostic sensitivity and the time of detection of MGIT 960 system were significantly higher than of Löwenstein-Jensen medium. PCR-RFLP identification analysis results were in concordance with those obtained by the conventional biochemical tests.
Abstract: A combination of Bactec MGIT 960 system, a PCR-based assay and a PCR-Restriction Analysis procedure (PRA) was assessed for the detection and identification of mycobacteria from clinical samples. The MGIT recovered 243 from 266 mycobacterial isolates. The PCR-based assay correctly identified all (234) Mycobacterium tuberculosis isolates, while 35 nontuberculous mycobacterial isolates were identified by PRA.
Abstract: A total of 300 Streptococcus pyogenes isolates, collected during 2001 from five hospitals in the Thessalia district (Central Greece), were examined for their resistance to macrolides. Resistance to erythromycin was detected in 58 isolates (19.3%). Of these, 68.9% were susceptible to clindamycin (M-phenotype) and carried the mefA gene. Of the remaining isolates, 18 expressed the MLS(B) phenotype: 12 and six exhibited inducible and constitutive resistance to clindamycin, respectively. All of these strains were found to be ermA(TR) positive, except for four that had the ermB gene. Of the erythromycin-resistant strains, none was found to be resistant to penicillin, tetracycline or quinupristin-dalfopristin. Molecular typing by PFGE showed the presence of a limited number of clones.
Abstract: One hundred and fifty clinical isolates of Mycobacterium tuberculosis were tested for susceptibility to pyrazinamide using the fully automated Bactec MGIT 960 system and the radiometric Bactec 460TB system. The overall concordance rate between MGIT 960 and radiometric system was 100% and the mean turnaround times to report the susceptibility test results were almost identical (6.37 and 6.8 days, respectively).
Abstract: /Aims: Laminin and collagen IV have been proposed as extracellular matrix serum markers. Because fibrosis is a major complication of inflammatory bowel disease, serum concentrations of laminin and collagen IV were measured in patients with ulcerative colitis (UC) and Crohn's disease (CD) and compared with inflammatory and healthy controls.
Abstract: Lung function in diabetes has been reported in several studies with contradictory results. Diabetes mellitus increases expression of adhesion molecules through hyperglycemia. These molecules play an important role in the pathophysiological dysfunction of the vasculature.
Abstract: A novel method, based on PCR Restriction Fragment Length Polymorphism Analysis (PRA) of a part of the tuf gene (370 bp), was designed for the identification of 11 staphylococcal species, including the most common staphylococcal pathogens. A total of 258 clinical isolates were validated by this assay, and the results were in concordance with those obtained by the reference method of Kloos and Schleifer.
Abstract: Oxacillin resistance was examined in 258 coagulase-negative staphylococci from Greek hospitals. mecA DNA was detected in 168 isolates, which were also resistant to oxacillin by agar dilution and disk diffusion, according to the current NCCLS breakpoints. Both methods exhibited a relatively low specificity misclassifying 21 and 19 of the 90 mecA-negative isolates respectively as oxacillin resistant. In contrast, an anti-PBP 2a latex agglutination test, applied after induction by oxacillin, correctly classified 163 mecA-positive (sensitivity 97%) and 88 mecA-negative isolates (specificity 97.7%).
Abstract: A dysregulated local immune defence with a constant influx of leucocytes provides a basis for continuous intestinal inflammation in ulcerative colitis and Crohn's disease. Since vascular adhesion protein 1 (VAP-1) is one of the adhesion molecules that mediates lymphocyte binding to endothelium, we investigated the levels of soluble VAP-1 (sVAP-1) in the sera of inflammatory bowel disease (IBD) patients compared with healthy controls.
Abstract: Matrix metalloproteases (MMPs) and their inhibitors are effector molecules involved in extracellular matrix remodelling. The serum profile for these proteolytic enzymes and their inhibitors during acute self-limiting viral hepatitis has not been studied. We therefore determined serum concentrations of MMP-1, MMP-3, MMP-2, MMP-9 and their inhibitors (tissue inhibitors of metalloproteinase) TIMP-1, TIMP-2 and alpha2 macroglobulin (AMG) in the serum of patients during the icteric stage of self-limiting acute viral hepatitis. Transforming growth factor-beta (TGF-beta) and interleukin (IL)-10, two cytokines involved in the regulation of MMPs and TIMPs were also assessed. Nineteen patients (12 men, seven women) with a mean age of 29.9 years (range 16-65 years) participated in the study. Fifteen had hepatitis B virus (HBV, two HCV and two HAV infection. The values of patients were compared with those obtained from 15 blood donor controls (eight men, seven women), mean age 36.2 years (range 18-55 years). Serum levels of TGF-beta, IL-10, MMP-1, MMP-3, MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed by ELISA. MMP-2 and MMP-9 were also measured by a zymogram protease assay. alpha2 macroglobulin (AMG) was measured by nephelometry. Compared with the healthy controls the mean serum concentrations of all MMPs were significantly decreased in the acute hepatitis patients. There was no difference in the serum concentration of TIMP-1 between patients and the controls. Serum levels of TIMP-2 (P < 0001), TGF-beta (P < 0.05), IL-10 (P < 0.001) and AMG (P < 0001) were increased in patients compared to healthy controls. A statistically significant negative correlation by linear regression analysis was found between AMG and MMP-1 (P=0003). The decreased levels of MMPs observed, together with normal and increased levels of TIMP-1 and TIMP-2, may indicate an attempt to limit matrix degradation at this stage of disease resolution. The increased levels of the anti-inflammatory cytokines IL-10 and TGF-beta might be the underlying mechanism responsible for the above effect. AMG inhibition especially for MMP-1 may play an additional important role.
Abstract: Detection of methicillin-resistant staphylococci is critical for the management of infected patients in the hospital. A total of 55 nonreplicated clinical isolates of staphylococci (31 Staphylococcus aureus and 24 coagulase-negative staphylococci; CNS) collected during a one-year period and expressing low-level resistance to methicillin (oxacillin MIC of 2-4 mg/l for S. aureus and 0.5-4 mg/l for CNS) were studied. mec determinants and overproduction of beta-lactamase were investigated and pulsed-field gel electrophoresis (PFGE) was applied as a typing method. Twenty-four S. aureus isolates and 19 CNS carried the mecA gene. The presence of mecR1/mecI and blaR1/blaI genes correlated with the expression of low-level methicillin resistance in CNS. Four mecA-negative isolates (2 S. aureus and 2 CNS) overproduced beta-lactamase. PFGE revealed the presence of 2 major clonal types in mecA-positive S. aureus isolates, and 3 in CNS. Low-level methicillin resistance of staphylococci is correlated with the presence of the mecA gene and overproduction of beta-lactamase.
Abstract: The distribution of the mec genes mecA, mecR1 and mecI that regulate the expression of methicillin resistance was investigated by PCR in 145 staphylococci of hospital origin. Determination of alterations and deletions in parts of the genes was achieved using 11 sets of primers in combined reactions. Methicillin-resistant Staphylococcus epidermidis strains appeared relatively stable, with 57.9% of isolates containing the whole regulatory region. Alterations within the mecA gene were detected more often in other coagulase-negative staphylococci, which also had a higher percentage with deletions of regulatory genes. Among methicillin-resistant S. aureus, a genetically heterogeneous population was identified, with several alterations and deletions of mec genes.
Abstract: A total of 250 consecutive Staphylococcus aureus clinical isolates were collected during the period 1999-2000 from the five major hospitals of the district of Thessaly (Central Greece). Thirty seven (14.8%) of the isolates were mecA-positive (MRSA) in a PCR-based assay; all exhibited resistance to oxacillin (agar dilution MICs > or =4 mg/L) and were also resistant to multiple antibiotics. Most of the MRSA isolates had been collected in the intensive care units and the surgical wards of the participating hospitals in a sporadic fashion. The MRSA incidence found here was significantly lower than reported in previous studies from Greece. Molecular typing by PFGE showed that the MRSA isolates were distributed between three pulsotypes. Evaluation of various conventional methods for assessing methicillin resistance showed that oxacillin agar dilution and immunological detection of PBP2a with the Slidex MRSA Detection kit were the most reliable in this setting. Misclassifications of isolates exhibiting low-level resistance (oxacillin MIC 2-4 mg/L) occurred with the salt agar screen, the oxacillin disk diffusion and the ATB Staph System methods.
Abstract: The combined measurement of perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) and anti-Saccharomyces cerevisiae mannan antibodies (ASCA) has recently been suggested as a valuable diagnostic approach in inflammatory bowel disease (IBD). The aim of this study was to assess the value of detecting pANCA and ASCA in the differentiation between ulcerative colitis (UC) and Crohn's disease (CD) in a Greek population with IBD.
Abstract: Analysis of the antibiotic binding capacity of penicillin-binding proteins (PBPs) in a group of clinical isolates of Staphylococcus epidermidis suggests that the increased level of resistance to methicillin (MIC 4.0 microg/ml) in an isolate free of the mecA gene is due to the decreased binding capacity of PBP3.
Abstract: A sample of 450 consecutive, non-replicated coagulase-negative staphylococci (CoNS), collected from clinical specimens during the period 2000-2001 from the five major hospitals of Thessaly district (Central Greece) were investigated for resistance to methicillin. Most of the isolates had been collected in a sporadic fashion from the intensive care units and the surgical wards of the participating hospitals. The majority of the isolates (76%) were Staphylococcus epidermidis (50%), Staphylococcus haemolyticus (14.8%) and Staphylococcus hominis (11.1%). All 316 isolates (70%) were classified as resistant according to NCCLS breakpoints (MIC > or =0.5 mg/l); 268 (59.5%) of them were mecA-positive in a PCR-based assay. All isolates with MIC > or =8 mg/l carried the gene, while, only 23.8% of isolates with MIC, 0.5-4 mg/l were carriers. Only 9% of the mecA-positive isolates were found to be sensitive to various non-beta-lactams, while 41.8% of the isolates were resistant to more than three antimicrobial groups apart from beta-lactams. Molecular typing by PFGE showed apparent heterogeneity among isolates of each species and the absence of predominant clones.
Abstract: Primary biliary cirrhosis and ulcerative colitis are two diseases with many features of autoimmunity. Thirteen cases of coexistence of the two diseases have been reported in the literature so far. Patients are usually younger and more often males than the ordinary primary biliary cirrhosis patient, while the colitis is mild and easily controllable. In a homogeneous population of 550,000 inhabitants of the island of Crete, 412 cases of ulcerative colitis and 82 individuals with primary biliary cirrhosis or autoimmune cholangitis have been identified. In two cases, coexistence of the two diseases was found. Immunological screening for AMA positivity in 150 ulcerative colitis sera disclosed no further cases. Prevalence of primary biliary cirrhosis in ulcerative colitis patients seems at least 30 times higher than in the general population in our area. A possible immunological link between the two diseases is discussed.
Abstract: Common aetiopathogenic factors may explain the association of ulcerative colitis with autoimmune disorders such as systemic lupus erythematosus.
Abstract: Fast tissue regeneration after therapeutic manipulations is a central problem of periodontology, oral surgery and trauma of the periodontal tissues, including bone. Several products, which augment tissue regeneration, have been manufactured and assayed in clinical practice with positive results. Emdogain is a recent addition in this field, as a tissue-regenerating product. The substance is a derivative of amelogenin, obtained from porcine embryonic tissues. At the present time, it is not known whether the substance can induce a local (due to the uptake of the substance) or systemic immune response. The aim of the present study was to evaluate, in vitro, the ability of Emdogain to influence, in vitro, the immune system. Peripheral blood lymphocytes, isolated for 10 healthy donors, were cultured in the presence of various concentrations of the substance, in order to determine the rate of cell proliferation, the expression of surface antigens and the production of cytokines and immunoglobulins. Under our experimental conditions, Emdogain produced a slight increase of the proliferation of lymphocytes, restricted to the CD25 (IL-2 receptor) fraction of the CD4 positive T-lymphocytes, and a concomitant decrease of CD19 positive B-lymphocytes. Other cell fractions (CD8 positive T-cells, B-cells and NK-cells) were not affected. Under our conditions too, immunoglobulin and cytokin (IL-2 and IL-6) production was not modified, even after a 3-day application of concentrations much higher than those used in clinical practice. Our data suggest that Emdogain slightly induce an immune response, restricted to the activated fraction of CD4 T-lymphocytes in vitro.
Abstract: Patients with inflammatory bowel disease (IBD) frequently suffer from thromboembolic events. Anti-cardiolipin (aCL) antibodies have been shown to be associated with thrombosis. Recently, the antibodies against the anti-cardiolipin cofactor beta2-glycoprotein I (a(beta2)GPI) have been found with higher specificity for thrombosis. The presence of these antibodies was assessed in 128 patients with IBD [83 with ulcerative colitis (UC) and 45 with Crohn's disease (CD)] and 100 healthy controls (blood donors). Patients with UC and CD had a significantly higher prevalence of aCL (18.1% and 15.6%, respectively) than healthy controls (HC) (3%). Eleven IBD patients (8.6%) but no HC had a(beta2)GPI. None of the IBD patients with a history of thrombosis had aCL and only one of them (a UC patient with deep vein thrombosis of the right leg) had a high titer of IgG a(beta2)GPI. In conclusion, these data show that both aCL and a(beta2)GPI are significantly associated with IBD but further studies are needed to determine the significance of our findings.
