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Aim: The immunosuppressive drug tacrolimus has several effects on the central nervous system. Besides its protective effect in hearing deficiencies, it is also considered to be able to cause tinnitus. In the present work, we attempted to describe its effects on a characteristic synapse of the auditory system that may be involved in the pathogenesis of tinnitus., Methods/materials: Slices of the dorsal cochlear nucleus (200 microm thick) were prepared from 9- to 14-day-old Wistar rats. In response to stimulation targeting the superficial layer of the nucleus, we recorded excitatory postsynaptic currents (EPSCs) developing in the cell bodies of the pyramidal neurons using whole-cell voltage clamps. Inhibitory synaptic activity was inhibited by the application of bicuculline and strychnine. Short-term plasticity was investigated using high-frequency stimulation (50 Hz). Unambiguous identification of the investigated neurons was ensured by employing biocytin in the pipette solution, which allowed the confocal reconstruction of the cells after the functional measurements. A concentration of 1 micromol/L tacrolimus was applied extracellularly., Results: Tacrolimus effectively and reversibly inhibited glutamatergic neurotransmission in the investigated synapse from -145 +/- 26 pA to -55 +/- 15 pA (n = 7; P = .00928). In contrast, EPSC amplitudes without failures were not significantly reduced (from -153 +/- 26 pA to -131 +/- 23 pA) in the presence of tacrolimus, but there were increased failure numbers of synaptic transmission. These data suggested that application of tacrolimus produced a combined pre- and postsynaptic inhibition., Conclusion: Tacrolimus affected short-term synaptic plasticity in the rat dorsal cochlear nucleus. It was also capable of inhibiting the glutamatergic neurotransmission. These effects suggested that tacrolimus may have neuroprotective effects in this structure., (Copyright 2010 Elsevier Inc. All rights reserved.)

Background and Aims: Recent studies have shown that dietary red palm oil (RPO) supplementation improves functional recovery following ischaemia/reperfusion in isolated hearts. The main aim of this study was to investigate the effects of dietary RPO supplementation on myocardial infarct size after ischaemia/reperfusion injury. The effects of dietary RPO supplementation on matrix metalloproteinase-2 (MMP2) activation and PKB/Akt phosphorylation were also investigated., Materials and Methods: Male Wistar rats were divided into three groups and fed a standard rat chow diet (SRC), a SRC supplemented with RPO, or a SRC supplemented with sunflower oil (SFO), for a five week period, respectively. After the feeding period, hearts were excised and perfused on a Langendorff perfusion apparatus. Hearts were subjected to thirty minutes of normothermic global ischaemia and two hours of reperfusion. Infarct size was determined by triphenyltetrazolium chloride staining. Coronary effluent was collected for the first ten minutes of reperfusion in order to measure MMP2 activity by gelatin zymography., Results: Dietary RPO-supplementation decreased myocardial infarct size significantly when compared to the SRC-group and the SFO-supplemented group (9.1 +/- 1.0% versus 30.2 +/- 3.9% and 27.1 +/- 2.4% respectively). Both dietary RPO- and SFO-supplementation were able to decrease MMP2 activity when compared to the SRC fed group. PKB/Akt phosphorylation (Thr 308) was found to be significantly higher in the dietary RPO supplemented group when compared to the SFO supplemented group at 10 minutes into reperfusion. There was, however, no significant changes observed in ERK phosphorylation., Conclusions: Dietary RPO-supplementation was found to be more effective than SFO-supplementation in reducing myocardial infarct size after ischaemia/reperfusion injury. Both dietary RPO and SFO were able to reduce MMP2 activity, which suggests that MMP2 activity does not play a major role in protection offered by RPO. PKB/Akt phosphorylation may, however, be involved in RPO mediated protection.

Objective: Systemic sclerosis (SSc) is a connective tissue disease that is characterized by microvascular disease and tissue fibrosis. Progressive loss and irregular architecture of the small blood vessels are well characterized, but the potential involvement of the lymphatic vessel system has not been analyzed directly in SSc. This study was undertaken to assess whether the lymphatic vascular system is affected in SSc, and whether changes to the lymphatic vessels are associated with dystrophic changes and tissue damage in patients with SSc., Methods: Lymphatic endothelial cells in skin biopsy samples from patients with SSc and age- and sex-matched healthy volunteers were identified by staining for podoplanin and prox-1, both of which are specifically expressed in lymphatic endothelial cells but not in blood vascular endothelial cells. CD31 was used as a pan-endothelial cell marker. Statistical analyses were performed using Kruskal-Wallis, Mann-Whitney U, and Spearman's rank correlation tests., Results: The numbers of podoplanin- and prox-1-positive lymphatic vessels were significantly reduced in patients with SSc as compared with healthy individuals. The number of podoplanin-positive lymphatic precollector vessels was significantly lower in SSc patients with fingertip ulcers than in SSc patients without ulcers. Moreover, the number of lymphatic vessels correlated inversely with the number of fingertip ulcers at the time of biopsy and with the number of fingertip ulcers per year. The inverse correlation between lymphatic precollector vessel counts and fingertip ulcers remained significant after statistical adjustment for the blood vessel count, age, and modified Rodnan skin thickness score., Conclusion: These results demonstrate a severe reduction in the number of lymphatic capillaries and lymphatic precollector vessels in patients with SSc. Patients with decreased lymphatic vessel counts may be at particularly high risk of developing fingertip ulcers.

Introduction: Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases capable of degrading extracellular matrix, including the basement membrane. MMPs are associated with various physiological processes such as morphogenesis, angiogenesis, and tissue repair. Moreover, due to the novel non-matrix related intra- and extracellular targets of MMPs, dysregulation of MMP activity has been implicated in a number of acute and chronic pathological processes, such as arthritis, acute myocardial infarction, chronic heart failure, chronic obstructive pulmonary disease, inflammation, and cancer metastasis. MMPs are considered as viable drug targets in the therapy of the above diseases., Methods: For the development of selective MMP inhibitor molecules, reliable methods are necessary for target validation and lead development. Here, we discuss the major methods used for MMP assays, focusing on substrate zymography. We highlight some problems frequently encountered during sample preparations, electrophoresis, and data analysis of zymograms., Results and Discussion: Zymography is a widely used technique to study extracellular matrix-degrading enzymes, such as MMPs, from tissue extracts, cell cultures, serum or urine. This simple and sensitive technique identifies MMPs by the degradation of their substrate and by their molecular weight and therefore helps to understand the widespread role of MMPs in different pathologies and cellular pathways., (Copyright 2010 Elsevier Inc. All rights reserved.)

Purkinje-like cells (PLCs) of the cochlear nucleus (CN) are strongly calbindin positive neurones with unknown function. In the present work functional and morphological methods have been employed to provide data about PLCs in general, and about their possible involvement in the synaptic organisation of the CN in particular. PLCs had slightly elongated soma, from which a complex dendritic arborisation extended with highly variable dimensions. On the basis of their morphology, three classes of PLCs were identified. Positively identified PLCs fired a train of action potentials on sustained depolarization. When hyperpolarizing stimuli were applied, the presence of a slowly activating, ZD7288-sensitive inward current was noted that corresponded to the h-current. PLCs received both excitatory and inhibitory synaptic inputs. Functional experiments revealed that 76% and 14% of the spontaneous inhibitory postsynaptic currents recorded from the cell bodies of the PLCs were mediated via glycinergic and GABAergic synapses, respectively. PLCs presented strong cerebellin1-like immunoreactivity, but its distribution differed from that seen in cerebellar Purkinje cells. Our results indicate that PLCs are parts of the synaptic circuitry of the CN, thus they may be actively involved in the processing and analysis of auditory information.

The involvement of astrocytes in the cholinergic modulation of the cochlear nucleus has been studied using primary astrocyte cultures prepared from this nucleus. The cells were loaded with the membrane permeable form of the fluorescent Ca(2+) indicator Fluo-4, and carbachol-induced Ca(2+) concentration increases were monitored using an imaging system. In the presence of cholinergic stimulation 36.3% of the cells produced Ca(2+) transients. The time course of the transients was variable; 45.0% of the responding cells showed only a rapid Ca(2+) concentration increase, while in 50.5% of the astrocytes the fast component was followed by a slow plateau phase. Using muscarine as well as general and more specific cholinergic antagonists (atropine, pirenzepine, 4-DAMP and hexamethonium), the role of the M3 and (to a smaller extent) M1 muscarinic acetylcholine receptors could be demonstrated in the genesis of the carbachol-induced Ca(2+) transients. The presence of these two subtypes of muscarinic receptors has been confirmed at both mRNA (Q-PCR) and protein (immunocytochemistry) levels. Our data demonstrate the responsiveness of the cochlear astrocytes towards cholinergic stimulation, suggesting that they may have roles in mediating the effects of cholinergic modulation in the rat cochlear nucleus.

Subclavian artery puncture and dilation with 9 Fr dilator during pacemaker implantation is a rare complication. We present a case, where this complication occurred and the injury was sealed with collagen-based closure device without any complications. Long term follow-up and the control angio-CT revealed no further late vascular complications. Therefore, use of a collagen based closure device for treating accidental iatrogenic subclavian artery injury can be a simple, successful alternative therapy when surgical direct vascular repair is not possible or not available.

Objective: To assess the efficacy of leflunomide, intravenous immunoglobulins, and ciprofloxacin as active treatment of postrenal transplant BK virus nephropathy (BKVN) in graft outcome at 1 year., Patients and Methods: Renal transplant recipients with positive results of 2 BK virus polymerase chain reaction tests of urine and blood underwent graft biopsy to confirm BKVN. If BKVN was diagnosed, antimetabolite therapy (mycophenolate mofetil or azathioprine) was changed to leflunomide therapy accompanied by a course of immunoglobulin and oral ciproflxacin., Results: Of 18 patients evaluated, 72% were men. Nine patients received cadaveric organs, with a mean of 3.6 HLA mismatches. All patients received induction thereapy (61% thymoglobulin), and 61% received antirejection therapy before BKVN was diagnosed. Maintenance immunosuppression therapy was primarily with prednisolone (94%); mycophenolate mofetil, 2 g/d (94%); and tacrolimus (61%). At baseline, mean (SD) creatinine clearance was 35.6 (11.5) mL/min/1.73(2), which decreased to 29.3 (17.3) mL/min/1.73(2) at 1 year (P = .01). Patients were divided into 2 groups of 9 each according to creatinine clearance values. In group 1, baseline value was 44.5 (6.6) mL/min/1.73(2), compared with 25.36 (7.8) mL/min/1.73(2) in group 2, which decreased to 42.66 (12.8) mL/min/1.73(2) (P = .23) and 16.76 (9.0) mL/min/1.73(2) (P = .009), respectively, at 1 year. Three grafts (16.7%) were lost by the end of the study, all in group 2 (P = .03)., Conclusion: Late diagnosis and intensive immunosuppression predispose to BKVN. Early active treatment of BKVN may improve graft outcome at 1 year posttransplantation.

Although cholinergic modulation of the cochlear nucleus (CN) is functionally important, neither its cellular consequences nor the types of receptors conveying it are precisely known. The aim of this work was to characterise the cholinergic effects on giant cells of the CN, using electrophysiology and quantitative polymerase chain reaction. Application of the cholinergic agonist carbachol increased the spontaneous activity of the giant cells; which was partly the consequence of the reduction in a K(+) conductance. This effect was mediated via M4 and M3 receptors. Cholinergic modulation also affected the synaptic transmission targeting the giant cells. Excitatory synaptic currents evoked by the stimulation of the superficial and deep regions of the CN were sensitive to cholinergic modulation: the amplitude of the first postsynaptic current was reduced, and the short-term depression was also altered. These changes were mediated via M3 receptors alone and via the combination of M4, M2 and M3 receptors, when the superficial and deep layers, respectively, were activated. Inhibitory synaptic currents evoked from the superficial layer showed short-term depression, but they were unaffected by carbachol. In contrast, inhibitory currents triggered by the activation of the deep parts exhibited no significant short-term depression, but they were highly sensitive to cholinergic activation, which was mediated via M3 receptors. Our results indicate that pre- and postsynaptic muscarinic receptors mediate cholinergic modulation on giant cells. The present findings shed light on the cellular mechanisms of a tonic cholinergic modulation in the CN, which may become particularly important in evoking contralateral excitatory responses under certain pathological conditions.

Vaccination is the main strategy to control severe dehydrating gastroenteritis caused by rotaviruses in early childhood. The availability of new generation rotavirus vaccines has led to an intensification of strain surveillance worldwide, in part, to gauge the impact of the possible vaccine-driven immune selection of wild-type rotavirus strains. In the present study, authors describe the strain prevalence data obtained in 2007, with the involvement of different regions of Hungary. Genomic RNA was extracted from rotavirus-positive stool samples collected mainly from children and then subjected to genotyping using multiplex RT-PCR assay. Type-specific primers targeted G1 to G4, G6, G8 to G10, and G12 VP7 specificities, and P[4], P[6], and P[8] to P[11] VP4 specificities were used. Out of 489 rotavirus-positive specimens, collected from 482 patients, 466 and 474 were successfully G and P typed, respectively, and both G and P type specificities could be assigned for 457 strains. Prevalence data showed the predominance of G4P[8] (31.5%) strains, followed by G1P[8] (28.3%), G2P[4] (19.3%), and G9P[8] (10.2%). Minority strains were G1P[4] (0.4%), G2P[8] (1.3%), G3P[9] (0.2%), G4P[6] (0.7%), G6P[9] (0.4%), G8P[8] (0.2%), G9P[4] (0.2%), G9P[6] (0.8%), and G12P[8] (0.4%). Mixed infections were found in 1.2% of the samples, while 4.9% remained partially or fully non-typified. Our data indicate that the antigen specificities of medically important rotavirus strains identified in this 1-year study are well represented in the vaccines available in the pharmaceutical private market in Hungary. Depending on the vaccination coverage achievable in the forthcoming years, the post-vaccination rotavirus strain surveillance may allow us to gain comprehensive information on the impact of rotavirus vaccines on the prevalence of circulating rotavirus strains.

The incidence and type distribution of enteric human adenoviruses (HAds) among diarrheic children in south-western Hungary was investigated from 2003 through 2006. Laboratory studies were conducted using commercial antigen detection tests (latex agglutination or immunochromatography), polymerase chain reaction (PCR) amplification, single-strand conformation polymorphism, and sequencing and phylogenetic analysis of a conservative region of the HAd hexon gene. The overall rate of HAd infection in childhood gastroenteritis cases during the 4-year study was 8.1%, with a gradual decrease in detection rates from 11.7% in 2003 to 5.7% in 2006. Molecular studies of a subset of HAd-positive samples found that enteric HAd type 40 strains were identified only in 2003 and 2004, while HAd type 41 strains were identified throughout the 4-year study. Higher detection rates of non-enteric HAds was documented during the first half of the study period when latex agglutination was used in our laboratory for detection. Our study suggests that the choice of diagnostic method may profoundly influence the epidemiologic picture and disease burden attributed to enteric HAd infections.

The purpose of this study was to investigate plasma homocysteine (Hcy) levels in patients with systemic sclerosis (SSc) and to study the association between plasma Hcy, C677T polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR), and the clinical manifestations in SSc. Associations of Hcy level, C677T MTHFR polymorphism, and macrovascular diseases were investigated in 152 patients with SSc and 58 controls. No significant differences in Hcy levels and MTHFR genotypes were found in SSc patients compared to controls or in SSc patients with limited cutaneous compared to diffuse disease. Significantly higher Hcy concentration was observed in patients with macroangiopathy/thromboembolic events compared to patients without such clinical manifestations (p < 0.05). There was significant correlation between age and macrovascular disorders, between Hcy level and the disease duration (r = 0.164; p < 0.05). Seventy-one percent of patients with macrovascular disorders had MTHFR polymorphism. In addition, 45% of patients with hyperhomocysteinemia had pulmonary hypertension. The presence of MTHFR C677T mutation influences the incidence of macrovascular abnormalities in SSc patients. Elevated Hcy levels may be associated with disease duration and the evolution of macrovascular disorders and pulmonary hypertension in SSc.

Human adenovirus (HAdV) serotype 52 has recently been discovered in the United States in samples from human patients with gastroenteritis of unknown etiology and is suspected to be a new human enteric pathogen. The aim of the present pilot study was to investigate whether this virus is circulating in the population of Southern Hungary by screening stool specimens collected from gastroenteritis cases and communal sewage samples in the area of Baranya County. A total of 209 diarrheic stool (124 from children and 85 from adults) and 45 influent sewage samples were screened for HAdV-52 by PCR using a primer pair specific to the gene of 12.5K protein in the E3 genomic region. The novel human adenovirus was not detected in any of the tested samples, suggesting that HAdV-52 was not circulating in the target population and the area during the study period. Since temporal and geographical fluctuations may markedly affect the epidemiology of human enteric pathogens, additional investigations are required to gain more in-depth insights into the ecology of this novel adenovirus.

Background: Hepatitis E virus (HEV) is a common cause of acute, fecally transmitted hepatitis in developing countries. Identification of HEV in indigenous human infection and in domestic pig raising the possibility that HEV infection is also a zoonosis., Objectives/study Design: Molecular detection and epidemiology of HEV in humans (South-East Hungary) with acute hepatitis and in domestic (pig, cattle) and wild (boar and roe-deer) animals (countrywide) by ELISA and RT-PCR., Results: Between 2001 and 2006, a total of 116 (9.6%) of 1203 human sera were positive by HEV IgM ELISA and 13 (24.5%) of 53 samples were also confirmed by RT-PCR and sequencing. Forty-two (27.3%) of 154, 11 (34.4%) of 32 and 9 (12.2%) of 74 samples were RT-PCR-positive from swine (feces: 22.7%; liver: 30.8%), roe-deer (liver) and wild boar (liver), respectively. Except for an imported infection caused by genotype 1, 19 sequences (human: 12, swine: 4, roe-deer: 1, wild boar: 2) belong to genotype 3 HEV. Genetically identical strains were detected in human and roe-deer and in 2 other human clusters., Conclusions: HEV is an endemic agent in Hungary. Consumption of raw or undercooked meat-products is one of the possible sources of the indigenous HEV infections. Cross-species infection with genotype 3 HEV potentially involves a food-borne transmission route in Hungary.

The spiral ganglion cells provide the afferent innervation of the hair cells of the organ of Corti. Ninety-five percent of these cells (termed type I spiral ganglion neurones) are in synaptic contact with the inner hair cells, whereas about 5% of them are type II cells, which are responsible for the sensory innervation of the outer hair cells. To understand the function of the spiral ganglion neurones, it is important to explore their membrane properties, understand their activity patterns and describe the variety of ionic channels determining their behaviour. In this review, a brief description is given of the various experimental methods that allow the investigation of the spiral ganglion cells, followed by the discussion of their action potential firing patterns and ionic conductances. The presence, distribution and significance of the K(+) currents of the spiral ganglion cells are specifically addressed, along with the introduction of the putative subunit compositions of the relevant voltage-gated K(+) channels.

Background: The importance of diarrhoeagenic Escherichia coli (DEC) infections in the Arabian Gulf including Kuwait is not known. The prevalence of DEC (enterotoxigenic [ETEC], enteropathogenic [EPEC], enteroinvasive [EIEC], enterohemorrhagic [EHEC] and enteroaggregative [EAEC]) was studied in 537 children < or = 5 years old hospitalised with acute diarrhoea and 113 matched controls from two hospitals during 2005-07 by PCR assays using E. coli colony pools., Results: The prevalence of DEC varied from 0.75% for EHEC to 8.4% for EPEC (mostly atypical variety) in diarrhoeal children with no significant differences compared to that in control children (P values 0.15 to 1.00). Twenty-seven EPEC isolates studied mostly belonged to non-traditional serotypes and possessed beta and theta intimin subtypes. A total of 54 DEC isolates from diarrhoeal children and 4 from controls studied for antimicrobial susceptibility showed resistance for older antimicrobials, ampicillin (0 to 100%), tetracycline (33 to 100%) and trimethoprim (22.2 to 100%); 43.1% of the isolates were multidrug-resistant (resistant to 3 or more agents). Six (10.4%) DEC isolates produced extended spectrum beta-lactamases and possessed genetic elements (blaCTX-M, blaTEM and ISEcp1) associated with them., Conclusion: We speculate that the lack of significant association of DEC with diarrhoea in children in Kuwait compared to countries surrounding the Arabian Gulf Region may be attributable to high environmental and food hygiene due to high disposable income in Kuwait.

EuroRotaNet was launched to monitor rotavirus strain prevalence during and after introduction of rotavirus vaccines in Europe. In early 2007, we detected P[6],G9 rotaviruses to appear in Hungary, representing the first documented occurrence of this strain in our surveillance area. Epidemiologic data suggested that this strain was introduced from India.

Unlabelled: Hepatitis E virus (HEV) is one of a common cause of acute, fecally transmitted hepatitis in developing countries. Identification of HEV in indigenous human infection and in domestic pig raises the possibility that HEV infection is also a zoonosis., Aim/methods: Molecular detection and epidemiology of HEV in humans with acute hepatitis and in domestic (pig, cattle) and wild (boar and roe-deer) animals by ELISA and RT-PCR in Hungary., Results: Between 2001 and 2006, a total of 116 (9.6%) human sera were positive by HEV IgM ELISA and 13 (24.5%) of 53 samples were also confirmed by RT-PCR and sequencing. Forty-two, 11 and 9 samples were RT-PCR-positive from swine (feces: 22.7%; liver: 30.8%), roe-deer (liver: 34.4%) and wild boar (liver: 12.2%), respectively. Except for an imported infection caused by genotype 1, 19 sequences (human: 12, swine: 4, roe-deer: 1, wild boar: 2) belong to genotype 3 HEV. Genetically identical strains were detected in human and roe-deer and in 2 other human clusters., Conclusions: HEV is an endemic agent in Hungary. Consumption of raw or undercooked meat-products is one of the possible sources of the indigenous HEV infections. Cross-species infection with genotype 3 HEV involves a food-borne transmission route in Hungary.

Although the hyperpolarization-activated non-specific cationic current (I(h)) plays important roles in determining the membrane characteristics of the spiral ganglion cells (SGCs), neither the exact types of the hyperpolarization-activated, cyclic nucleotide-gated, cation non-selective channel (HCN) subunits contributing to the molecular assembly of the relevant channels, nor their distribution pattern presented by the SGCs is known. In the present work immunolabeling and Western blot analysis were performed to describe the presence and distribution of all four known HCN subunits in the guinea-pig spiral ganglion. Besides determining the expression of the HCN1-HCN4 subunits by both type I and type II SGCs, the presence of possible apico-basal gradients in the expression patterns was also sought. The results indicate that both type I and type II SGCs express all four HCN subunits. The intensity of the immunolabeling of the cell surface membrane was generally strong, but it showed pronounced cell-to-cell variability. The Western blot experiments in combination with densitometry revealed that the amount of the HCN1 and HCN3 proteins was more significant in the apical than in the basal third of the guinea-pig cochlea. These findings not only imply potential heteromeric HCN channel formation of the spiral ganglion neurons, but they also offer a possible explanation of the previously reported heterogeneity of I(h) recorded in functional studies.

Rotavirus strain surveillance is being conducted in many countries before and after introduction of newly licensed vaccines to assess the impact of the vaccines on rotavirus strains. Here we describe a strain surveillance study in the Budapest area of Hungary (2003-2006) based on RNA profile analysis, genotyping by multiplex PCR and nucleotide sequencing. Among 1,983 G-typed rotaviruses we identified G1 (22%), G2 (4.8%), G3 (3.5%), G4 (18.5%), G6 (1.1%), G8 (<0.1%, n = 1), G9 (42%), and G12 (3.4%) specificities. Information on P genotype incidence was determined for a subset of samples (n = 814). In addition to the globally important strains, a variety of uncommon antigen combinations were also found, for example, P[9],G3; P[14],G6; or P[14],G8. Sequence and phylogenetic analysis of the VP7, VP4, VP6, and NSP4 genes of selected strains with uncommon antigen combinations demonstrated high similarity with certain bovine, porcine, feline, equine, and lapine rotaviruses, respectively. Continued surveillance is needed to assess the role of animal rotaviruses in human diseases., ((c) 2008 Wiley-Liss, Inc.)

Endocardial radiofrequency catheter ablation was performed because of frequent, 150/min monomorphic ventricular tachycardia (VT). In February 2005 it was many times terminated only by ICD shock in a 55-year-old postinfarction patient who received an implantable cardioverter defibrillator (ICD) 5 years ago because of rapid, monomorphic VT. After being asymptomatic for two months, a slower, 120/min, however, incessant ventricular tachycardia was present which was untreatable by a repeated endocardial ablation. Combined antiarrhythmic treatment was not effective either. In June 2005, after another unsuccessful endocardial ablation, epicardial ablation was decided as an "ultimum refugium". After subxyphoidal percutaneous pericardial punction we positioned the ablation catheter in the pericardial space, and ablation at the earliest activation point terminated the permanently ongoing arrhythmia for one and a half month within 5 seconds. After another three ablations we were unable to induce ventricular arrhythmia even with programmed ventricular extrastimulation. During a three-year follow-up, ventricular tachycardia was noticed in only two cases, ICD terminated both arrhythmias with the first antitachycardia pacing. The patient is in NYHA stage II at present. According to our knowledge, our case is the first successful epicardial ablation of incessant ventricular tachycardia in a postinfarction patient in Hungary.

Background: Some tumor-associated antigens (TAAs) are expressed on inflammatory cells. We previously detected increased production of CA15-3, CA19-9 and CA125 in rheumatoid arthritis (RA). The production of some TAAs may also be increased in patients with systemic sclerosis (SSc), systemic lupus erythematosus (SLE) and other connective tissue diseases. Some of these TAAs contain sialylated carbohydrate motifs and they are involved in tumor-associated cell adhesion and metastasis., Objectives: We assessed levels of TAAs in the sera of SSc, SLE patients, patients with infectious diseases and healthy subjects. Serum TAA levels were correlated with each other, as well as with disease activity markers and organ involvement., Methods: TAAs including CEA, CA15-3, CA72-4, CA125 and CA19-9 were assessed by immunoassay in the sera of 92 patients with SSc, 40 patients with SLE, 50 age- and sex-matched healthy controls, as well as with 40 patients with current bacterial or viral infections. Normal upper limits for these TAAs were 3.4 mg/l, 25 kU/l, 6.9 kU/l, 35 kU/l and 34 kU/l, respectively., Results: There were significantly more SSc patients showing abnormally high levels of CA19-9 (8.8% vs 2.0%), CA125 (11.0% vs 6.0%) and CA15-3 (28.4% vs 14.0%) in comparison to controls (p < 0.05). In SLE, significantly more patients had elevated levels of CEA (32.5% vs 20.0%), CA19-9 (7.5% vs 2.0%), CA125 (15.0% vs 6.0%) and CA72-4 (15.0% vs 8.0%) than did controls (p < 0.05). The mean absolute serum levels of CEA (6.6+/-1.7 vs 1.8+/-1.4 mg/l) and CA15-3 (22.9 +/- 1.8 vs 18.6 +/- 2.2 kU/l) were also significantly higher in SSc compared to controls (p < 0.05). We found numerous correlations between the serum levels of different TAAs within the SSc and SLE population. Among SSc patients, serum CEA (R = 0.290; p = 0.005), CA15-3 (R = 0.260; p = 0.020) and CA19-9 (R = 0.257; p = 0.013) correlated with renal involvement. Serum CA15-3 also correlated with joint involvement (R = 0.329; p = 0.003), ANA positivity (R = 0.288; p = 0.010) and CRP levels (R = 0.407; p < 0.001). Within the SLE population, serum CA72-4 correlated with central nervous involvement (R = 0.624; p = 0.004) and CA125 correlated with the SLEDAI composite activity index (R = 0.666; p = 0.002). Patients with infections exerted serum TAA patterns similar to healthy controls., Conclusion: The concentration of some TAAs may be elevated in the sera of patients with SSc or SLE in comparison to healthy subjects. Pathogenically, most of these TAAs contain carbohydrate motifs and thus they may be involved in inflammation-associated adhesive events. Furthermore, the production of some TAAs may correlate with organ involvement or disease activity in scleroderma or lupus.

Aims: Phrenic nerve stimulation (PNS), which is often intolerable for the patient, is a known complication of resynchronization therapy. We describe a new, minimal invasive method for treating PNS., Methods and Results: Untreatable PNS was found in nine cardiac resynchronization therapy patients with distal coronary sinus (CS) lead position 6 +/- 6 (0.5-17) months after the implantation. Ablation catheter and Amplatz Left 2 type guiding catheter were introduced into the right atrium via the right femoral vein. Coronary sinus was cannulated with the Amplatz catheter, and on a normal guide wire, a coronary stent was introduced beside the lead into the side branch in seven cases or a bigger stent into the CS in two patients. The ablation catheter was looped around the CS lead in the atrium with bent tip and was drawn backward together with the CS electrode. New lead positions were evaluated with electrophysiological measurements, and the suitable position was stabilized with inflation of the stent. Pericardial effusion was not detected on post-operative echocardiography. After repositioning, suitable pacing parameters were registered (threshold: 1.6 +/- 1.1 V; 0.5 ms, impedance: 565 +/- 62 ohm). Phrenic nerve stimulation was not found with 7.5 V; 1.5 ms pacing. During follow-up (7.7 +/- 4.6 months), stable pacing threshold and impedance values were measured; transient and reprogrammable PNS was present in only one patient., Conclusion: Coronary sinus electrode reposition using the femoral approach seems to be a safe and effective procedure, which means smaller burden for the patients compared with the established reposition operation. The technique can be used successfully if the CS lead is in a distal position.

Recent results suggest that B cells may have multiple pathogenic roles in systemic sclerosis (SSc) and there may be increased incidence of B cell lymphomas in SSc. Here, we assessed the prevalence of lymphomas in a large SSc cohort. We analyzed data of 218 Hungarian patients undergoing follow-ups in our institutions between 1995 and 2007. During this follow-up period, there were three SSc patients, who eventually developed B cell lymphoma. The first case is a woman with diffuse cutaneous form of SSc (dcSSc) including pulmonary, cardiac, gastrointestinal, and renal manifestations and anti-topoisomerase I antibody positivity. B cell chronic lymphocytic leukemia (B-CLL) with Zap70 expression (Rai I stage) developed 2 years after the onset of SSc. The second case is a woman with dcSSc presenting with pulmonary, cardiac, and gastroesophageal manifestations. Twenty-one months after disease onset, a chronic small lymphocytic B cell non-Hodgkin's lymphoma was diagnosed from retroperitoneal lymph nodes. Our third case is a woman with dcSSc and no internal organ manifestations. She also developed Zap70-positive B-CLL, stage Rai I 9 months after the onset of SSc. Thus, there were three cases of B cell lymphoma among our 218 SSc patients (1.38%). The association of scleroderma and non-Hodgkin's lymphoma may be a rather uncommon feature; however, the incidence of lymphoma among Hungarian SSc patients may be 1.9-2.5 times higher than that in the general population. In our three patients, B cell lymphoma developed within 2 years after the onset of SSc. Altered B cell function implicated in the pathogenesis of SSc may lead to the development of lymphoid malignancies.

Objective: Rho-associated kinases (Rock) are the major cellular mediators of Rho GTPases and play an important role in the organization of the actin cytoskeleton. Inhibitors of Rock are currently being evaluated for the treatment of pulmonary arterial hypertension. This study was undertaken to analyze the role of Rock in the activation of fibroblasts in systemic sclerosis (SSc)., Methods: Rock signaling was inhibited using chemical inhibitors and small interfering RNA. The expression of extracellular matrix (ECM) proteins and alpha-smooth muscle actin was analyzed by real-time polymerase chain reaction, Western blotting, and SirCol assay. Metabolic activity was quantified by MTT assay. Cell viability was assessed by staining with annexin V and propidium iodide. The role of MAP kinases was investigated using selective inhibitors and Western blotting., Results: Inhibition of Rock strongly reduced the synthesis of the major ECM proteins at the messenger RNA level as well as the protein level. Counterregulatory changes in the expression of tissue inhibitors of metalloproteinases and matrix metalloproteinases were not observed. Inhibition of Rock prevented myofibroblast differentiation. Transforming growth factor beta activated ERK in a Rock-dependent manner, and ERK mediated in part the stimulatory effects of Rock on myofibroblast differentiation. Toxic adverse effects of the inhibition of Rock were not observed., Conclusion: Our findings demonstrate that Rock potently stimulates the differentiation of resting fibroblasts into myofibroblasts and the production of ECM at biologically relevant concentrations without cell toxicity. These findings, along with the beneficial effects of Rock inhibition on vascular disease, indicate that inhibition of Rock might be an interesting novel therapeutic approach for the treatment of SSc.

Group C rotaviruses have been recognized as a cause of acute gastroenteritis in humans, cattle, and swine, although the true epidemiologic and clinical importance of this virus in these hosts has not yet been fully established. A real-time PCR assay based on a broadly reactive primer pair was developed and used to quantitatively determine the viral load of group C rotaviruses in environmental samples. A total of 35 raw and 35 treated sewage samples collected at the same sampling time in four Hungarian sewage treatment plants during a survey in 2005 were tested for the presence of group C rotaviruses. The overall detection rates were 91% (32 of 35) for the influent and 57% (20 of 35) for the effluent samples. Molecular characterization of the amplified partial VP6 gene revealed the cocirculation of human and animal (i.e., bovine and porcine) strains that were easily distinguishable by melting curve analysis. Human strains yielded relatively high viral loads (mean, 1.2 x 10(7); median, 6.9 x 10(5) genome equivalents per liter influent sewage) and appeared to display seasonal activity over the study period, whereas animal strains appeared to circulate throughout the year at much lower average titers (bovine strains mean, 9.9 x 10(4); median, 3.0 x 10(4); porcine strains mean, 3.9 x 10(4); median, 3.1 x 10(4) genome equivalents per liter influent sewage). Our findings suggest that monitoring of communal sewage may provide a good surrogate for investigating the epidemiology and ecology of group C rotaviruses in humans and animals.

Background: Noroviruses are common pathogens in human gastroenteritis outbreaks worldwide. They belong to a genetically diverse group of RNA viruses with multiple genogroups (G) and genotypes. Genotype GII-4 norovirus (Lordsdale) is the predominant agent in epidemics., Objectives: To investigate the genetic variation in GII-4 strains isolated during seven epidemic seasons in Hungary from November 2000 to June 2007., Study Design: Using the prospective epidemiological surveillance of norovirus outbreaks in Hungary, GII-4 strains were selected for further genetic analysis. After phylogenetic analysis, RNA-polymerase (open reading frame 1; ORF1), capsid (ORF2) and the ORF1/ORF2 junction were analysed by RT-PCR and sequencing., Results: Three hundred and seventy-seven (76.8%) of 491 confirmed norovirus outbreaks were caused by genotype GII-4. GII-4 was the predominant genotype in six of the seven epidemic seasons. Four main GII-4 variants--epidemic point mutants--(GII-4-2000, GII-4-2002, GII-4-2004 and GII-4-2006b) were detected, with each variant predominating in two consecutive epidemic seasons., Conclusions: Genotype GII-4 was confirmed as the predominant genetic type in epidemic norovirus seasons in Hungary. Genetic drift successfully promotes the re-emergence of GII-4 variants in the population. The elevated number of norovirus outbreaks in the population predicts the emergence of new GII-4 genetic variants as part of an international epidemic.

The spiral ganglion accommodates the cell bodies of the acoustic nerve fibres connecting the hair cells to the central nervous system. As the ionic channels containing various voltage-gated K+ channel (Kv) subunits play pivotal roles in determining the functional properties and firing behaviour of the spiral ganglion cells (SGCs), every piece of information concerning the Kv expression of the SGCs is valuable. In the present work a comprehensive immunohistochemical analysis was performed to describe the expression of 9 Kv subunits in the guinea pig cochlea on traditional wax-embedded sections as well as employing a newly developed preparation that allowed confocal analysis, reconstruction of the three-dimensional appearance and precise morphological characterisation of the SGCs. Besides determining their Kv expression patterns, differences between type I and type II SGCs were sought. SGCs showed positivity for 8 out of the 9 Kv subunit-specific antibodies with varying intensity and proportion of the immunopositive cells; whereas no obvious Kv3.2 positivity could be noted. Type I and type II cells demonstrated similar expression patterns for all subunits tested, with the exception of Kv1.2, whose presence was confirmed in only 50% of the type II cells. Although the present findings suggest that type I and type II cells do not differ fundamentally in the Kv subunits they possess; they also imply that SGCs may not form a homogeneous cell population, and might provide explanation of the previously noted heterogeneity of the membrane properties of the SGCs.

Skin vasculitis may be associated with infections and autoimmune diseases. Furthermore, vasculitis may appear as a paraneoplastic symptom. A 19-year-old male patient was examined with swollen joints and papules presented on lower extremitis. Laboratory results showed high erythrocyte sedimentation ratio, positive C reactive protein, increased number of leukocytes and high circulating immune complex level. Histopatology of skin biopsy specimen proved vasculitis. ANCA was not present. No other changes could be observed besides skin involvement. Symptoms disappeared on 0.5-mg/bwkg methylprednisolon therapy. Few weeks later, enlarged cervical lymph nodes developed besides fever and weight loss. Biopsy indicated the presence of mixed cell type Hodgkin lymphoma. Appropriate examinations revealed clinical stage III/B with favorable prognosis (IPS=2). After eight cycles of ABVD therapy complete remission was achieved as confirmed by FDG-PET. As a consequence of the treatment of Hodgkin lymphoma, vasculitis also disappeared. The present case report calls to attention the importance of careful examinations to exclude other diseases, especially malignancies that may remain at the background of cutaneous vasculitis. Treatment of the primary disease also results in the improvement of secondary skin vasculitis.

Because the neuronal membrane properties and firing characteristics are crucially affected by the depolarization-activated K(+) channel (Kv) subunits, data about the Kv distribution may provide useful information regarding the functionality of the neurons situated in the cochlear nucleus (CN). Using immunohistochemistry in free-floating slices, the distribution of seven Kv subunits was described in the rat CN. Positive labeling was observed for Kv1.1, 1.2, 1.6, 3.1, 3.4, 4.2, and 4.3 subunits. Giant and octopus neurons showed particularly strong immunopositivity for Kv3.1; octopus neurons showed intense Kv1.1- and 1.2-specific reactions also. In the latter case, an age-dependent change of the expression pattern was also documented; although both young and older animals produced definite labeling for Kv1.2, the intensity of the reaction increased in older animals and was accompanied with the translocation of the Kv1.2 subunits to the cell surface membrane. The granule cell layer exhibited strong Kv4.2-specific immunopositivity, and markedly Kv4.2-positive glomerular synapses were also seen. It was found that neither giant nor pyramidal cells were uniform in terms of their Kv expression patterns. Our data provide new information about the Kv expression of the CN and also suggest potential functional heterogeneity of the giant and pyramidal cells.

The presence of TASK-3 channels has been described in a number of healthy and malignantly transformed cells, showing mainly intracellular distribution with relatively insignificant labelling of the cell surface membrane. In this work, immunochemical and molecular biology methods were utilised to establish the intracellular organelle whose TASK-3 expression accounts for this strong intracellular labelling using cultured melanoma and HaCaT cells. Before the immunocytochemical experiments, the presence of TASK-3 mRNA was also confirmed in melanoma cells. Comparison of the results of the TASK-3- and mitochondrion-specific labelling indicated that the TASK-3 channel subunits were strongly expressed by mitochondria in both investigated cell types. Moreover, prominent TASK-3 expression of keratinocytes could also be demonstrated in histological sections excised from the human skin. These results indicate that TASK-3 channels are present in the mitochondria in both malignantly transformed and healthy cells, suggesting that they might have roles in ensuring mitochondrial functions.

Tula hantavirus (TULV) is a member of the genus Hantavirus, family Bunyaviridae and is mainly carried by the European common vole (Microtus arvalis). In order to detect TULV, we tested Microtus arvalis (MAR) and Microtus subterraneus (MSU) voles captured in two different locations of the Southern Transdanubian region of Hungary. The viral genome was detectable in 37% of the tested MAR voles but, interestingly, was absent in all MSU. Phylogenetic analysis performed with a partial coding sequence of the capsid gene showed that Hungarian TULV strains clustered with viruses detected in western Slovakia and in the Czech Republic. To the best of our knowledge, this is the first report on the identification of TULV detected in MAR voles in the Transdanubian region of Hungary.

Objective: Survival analysis of a series of 366 consecutive patients with systemic sclerosis (SSc)., Methods: Clinical and laboratory data were evaluated from 1983 until 2005 using a standard protocol. The female/male ratio was 315/51. The mean (SD) age of the patients was 56.8 (12.2) years. The duration of disease was 12 (5-19) years with a median follow-up of 6 (3-12) years., Results: Kaplan-Meier univariate analysis showed that renal, cardiac involvement, pigmentation disturbances, malabsorption, a forced vital capacity <50%, diffuse scleroderma, presence of early malignancy, anaemia, and increased erythrocyte sedimentation rate (ESR) were signs of unfavourable prognosis, whereas anti-centromere antibodies were indicators of a good survival. In the multivariate Cox proportional hazards model the presence of diffuse scleroderma, renal involvement, coexistence of a malignant disease, and increased ESR were poor independent prognostic signs. Elderly age at the onset of disease also caused an unfavourable outcome. A total of 86 SSc-related deaths were recorded during the follow-up. Of them, 65% were attributed to cardiorespiratory manifestation of disease. Tumour associated early death was found in 12 cases (14%)., Conclusions: In addition to the well-known factors influencing the outcome (diffuse subset, internal organ involvements, and inflammatory signs), the coexistence of scleroderma with a malignancy also causes a poor outcome.

There are no data on the serotypes of rotaviruses prevalent in Kuwait, which has a large expatriate population and hence a focal point for transmission of pathogens. The serotype information will contribute to the fund of knowledge on the world epidemiology of rotavirus serotypes and will predict the outcome of vaccination in Kuwait. Of the 75 rotavirus-positive samples from 172 children (aged <5 years) with severe diarrhoea, 69 were genotyped. The distribution of genotypes was G1 (63.8%) followed by G9 (10.2%), G2 (7.3%), G4 (7.3%) and G3 (4.4%). Among the P types, P[8] was the most common type found across all G types. By fluorescent focus neutralization test, serum antibodies to genotypes G1 (94%), G4 (68%) and G9 (46%) were found in 120 other children. These results show that G1 is the predominant serotype in Kuwait and that a vaccine that contains G1 will be most effective.

Group A rotaviruses are the most common cause of severe gastroenteritis worldwide. The incidence and distribution of group A rotavirus sero/genotypes varies between geographical areas during a rotavirus season, and from one season to the next. In addition, cocirculation of genetically diverse multitypic rotaviruses and of intratypic variants in any one place and time is common. Assuming widespread use of rotavirus vaccine in the near future, comprehensive surveillance of natural rotavirus infections is vital. EuroRotaNet has been established in order to gather comprehensive information on the rotavirus types co-circulating throughout Europe. The main objectives of the network are to (i) develop methods and algorithms for effective rotavirus strain typing and characterisation, (ii) describe in detail the molecular epidemiology of rotavirus infections in Europe, (iii) monitor the effectiveness of current genotyping methods and respond to changes associated with genetic drift and shift, and (iv) monitor the emergence and spread of novel rotavirus strains within Europe. This infrastructure may serve as a platform for future surveillance activities and nested studies for evaluating the effectiveness of a rotavirus vaccine in the general population. Studies to monitor the reduction in disease associated with common rotavirus types, the possible vaccine-induced emergence of antibody escape mutants of genotypes other than those included in the vaccine and of reassortment between vaccine and naturally circulating wildtype strains are required.

Dobrava hantavirus (DOBV) belongs to the genus Hantavirus of the family Bunyaviridae, and is carried by yellow necked and striped field mice (Apodemus flavicollis and Apodemus agrarius), respectively. The aim of this study was to detect and genetically characterize new DOBV strains in rodents captured in the Transdanubian region of Hungary. Rodent corpses were dissected and lung tissues were used for hantavirus detection by SYBR Green-based real-time RT-PCR using specific primers located in the S-segment of the virus genome. A total of 22 captured animals of the Apodemus species were tested for the presence of DOBV. Three out of the 22 mice were positive. Phylogenetic and molecular sequence analyses showed that Hungarian DOBVs were most closely related to those viruses detected from A. agrarius mice in Slovenia. Based on our new data from the region we concluded that extended reservoir studies would be necessary in the future.

The development of autoimmune diseases may be influenced by hormonal, immunomodulatory, and metabolic pathways. Prolactin (PRL), ferritin, vitamin D, and the tumor marker tissue polypeptide antigen (TPA) were measured in autoimmune diseases: systemic lupus erythematosus (SLE), systemic sclerosis (SSc), rheumatoid arthritis (RA), polymyositis (PM), dermatomyositis (DM), multiple sclerosis (MS), autoimmune thyroid diseases, and antiphospholipid syndrome. Hyperprolactinemia (HPRL) was detected in 24% of PM patients, in 21% of SLE patients, in 6.7% of MS patients, 6% of RA patients, and in 3% of SSc patients. Hyperferritinemia was detected in 23% of SLE patients, 15% of DM patients, 8% of MS patients, and 4% of RA patients. The patients had relatively low levels of 25 OH Vitamin D: the average results (mean +/- SD) were between 9.3 +/- 4.4 to 13.7 +/- 7.1 ng/mL in the different diseases, while the 25 OH Vitamin D concentrations less than 20 ng/mL are regarded as deficient. TPA levels were in the same range of the controls, elevated only in SLE. HPRL, hyperferritinemia, hypovitaminosis D, and TPA levels did not correlate with SLE activity elevated levels of rheumatoid factor or anti-CCP antibodies in RA. HPRL, hyperferritinemia, and hypovitaminosis D have different immunological implications in the pathogenesis of the autoimmune diseases. Preventive treatment with vitamin D or therapy for HPRL with dopamine agonists, may be considered in certain cases. Hyperferritinemia may be used as an acute-phase reactant marker in autoimmune diseases mainly SLE. TPA may be used to indicate the tendency for malignancies.

Introduction: Hepatitis A virus (HAV) is one of the most important cause of fecally transmitted acute infectious hepatitis worldwide. In Hungary, beside the sporadic HAV infections, outbreaks also occur, particularly in Northeast part of the country where the subgenotype IA is endemic. The reported number of HAV cases was less than 10 per year in Southwest Hungary., Aims: A part of the European HAV surveillance, the author's aims were to follow and to analyze the outbreak of hepatitis A in Transdanubia (Southwest Hungary) in 2006 by molecular epidemiological methods., Materials and Methods: Sera samples from symptomatic patients were tested prospectively by enzyme-immunoassay (EIA) and reverse transcription-polymerase chain reaction (RT-PCR) followed by sequence- and phylogenetic analysis., Results: Between June and December 2006, a total of 115 serum samples were positive for HAV IgM antibody in Southwest Hungary. Thirty (76.9%) of the 39 samples were RT-PCR-positive and contain genetically identical subgenotype IB hepatitis A virus (HAV/Transdanubia/2006/HUN) which has 100% nucleotide identity to strain IT-MAR-02 from Italy in 2002. Until now, approximately 1200-1300 persons have been infected with the probably imported strain. The average age of patients was 18 years (years 1 to 80). In this study, detailed clinical and epidemiological data of the outbreak are presented., Conclusions: Only a prospective molecular epidemiological study could verify the connection between the first sporadic HAV cases in June, the outbreak in August and the endemic spread of the virus since September in Southwest Hungary. This epidemic calls attention to how a hepatitis A outbreak can develop anywhere and at any time in Hungary and to the importance of active HAV immunization in the primary prevention.

Objectives: The pathogenesis of systemic sclerosis (SSc) includes vasculopathy with endothelial dysfunction. The aim of this study was to investigate endothelium-dependent, flow-mediated dilatation (FMD), as well as endothelium-independent, nitroglycerin-mediated dilatation (NMD) of the brachial artery and to assess common carotid intimal-medial thickness (ccIMT) in SSc patients compared with healthy controls., Methods: FMD and NMD of the brachial artery were determined using high-resolution ultrasound imaging and the values were expressed as percentage change from baseline in 29 SSc patients and 29 healthy controls. The two groups were very similar regarding sex, age and traditional cardiovascular risk factors. In addition, common carotid arteries were assessed by duplex colour ultrasound, ccIMT determined using high resolution ultrasound and expressed in mm thickness in the same patients and controls. Correlations between FMD, NMD, ccIMT, age and the SSc subtype (diffuse or limited form) were analysed., Results: In the 29 SSc patients (mean age: 51.8 yrs), the FMD was significantly lower (4.82 +/- 3.76%) in comparison with the controls (8.86 +/- 3.56%) (P < 0.001). No difference was found in NMD between patients (19.13 +/- 17.68%) and controls (13.13 +/- 10.40%) (P > 0.1). There was a tendency of increased ccIMT in SSc patients (0.67 +/- 0.26 mm) compared with healthy subjects (0.57 +/- 0.09), but this difference was not significant (P = 0.067). A significant, positive correlation between ccIMT and age in SSc (r = 0.470, P = 0.013) was detected, as well as in healthy controls (r = 0.61, P = 0.003), but no correlation was found between FMD and age. In addition, ccIMT, but not FMD and NMD, displayed significant correlation with disease duration (r = 0.472, P = 0.011). NMD displayed significant inverse correlation with the age in SSc patients (r = -0.492, P = 0.012), but not in controls. We did not find any correlation between FMD, NMD, ccIMT and SSc subtype., Conclusions: There is an impairment of endothelium-dependent vasodilatation indicated by low FMD in SSc. At the same time, the endothelium-independent dilatation assessed by NMD is still preserved giving an opportunity of nitroglycerine therapy. Carotid atherosclerosis indicated by ccIMT may occur at higher ages and after longer disease duration. Thus, the assessment of FMD in the pre-atherosclerotic stage may have a beneficial diagnostic, prognostic and therapeutic relevance.

Background: Soluble, human low affinity Fcgamma receptors, such as sFcgammaRII and sFcgammaRIII, are known to play a pathologic role in different diseases. Sandwich ELISAs had previously been applied for the specific detection and determination of these soluble receptors. In these ELISAs, commercial monoclonal antibodies (Ab) were used as capture antibodies with monoclonal or polyclonal antibodies serving as detector Abs. Increased levels of cell-free FcgammaRIII have been detected in patients with lupus but the functions and levels of sFcgammaRII have not been fully characterized yet., Objectives: The aim of this work was to determine the ligand binding capacities and levels of soluble FcgammaRII and FcgammaRIII in sera of patients with systemic lupus erythematosus (SLE). Moreover, correlation between the levels of sFcgammaRII and sFcgammaRIII and the clinical activity of the disease were investigated., Methods: Sera of 47 patients with SLE, and 51 healthy subjects were analyzed. In the newly developed indirect sandwich ELISAs commercial monoclonal anti-FcgammaRs are used as capture antibodies, and the ligand of FcgammaRII and FcgammaRIII, an artificial immune complex (IC), serves as a detection component replacing the second antibodies used in previous methods., Results: The ligand binding capacity of both soluble FcgammaRII and sFcgammaRIII were elevated in the sera of SLE patients compared to control samples. This increase was significant in patients with the active disease (n = 30; p < 0.01). It was also revealed that a substantial part of the soluble Fcgamma receptors in these patients was bound in vivo to circulating IC., Conclusion: These newly developed ELISAs are probably more phisiologically relevant than other previous assays because they detect the circulating receptors on the basis their in vitro ligan binding capacities. Therefore this method can separately measure the levels of the soluble, free FcgammaRs and those bound circulating IC in vivo.

Nowadays the management strategy for primary gastric lymphoma is undergoing change due to the effectiveness of chemotherapy and immunotherapy. While earlier surgical resection was the primary treatment and chemotherapy was only a follow-on procedure, at the present time this arrangement seems to have been reversed. Early stage low-grade MALT lymphoma can be treated with Helicobacter pylori eradication. Total or subtotal gastrectomy with D2 lymphadenectomy and with adjuvant chemotherapy in R1 situation is proposed up to stage II.1. The strategy is similar in the case of high-grade gastric lymphoma, but resection is useful only in those cases when one can be assured the result will be an R0 situation. With the exception of these cases, the only indication for resection is chemo-resistance. There is no reason for operating in advanced stages of the disease. The only purpose can be to manage complications. Unfortunately, the exact diagnosis is difficult. The diagnosis of lymphoma can often only be made after the operation. In the 6-year period between 1st January 2000 and 31st December 2005 we treated 38 patients for primary gastric lymphoma. Altogether 9 patients were operated on. Resection was performed in 6 cases. The diagnosis of lymphoma was known preoperatively only in one case. Nowadays surgery seems to be only secondary to chemotherapy and immuno-chemotherapy in the treatment of primary gastric MALT lymphoma. Our own cases also mirror this change.

Adequate interpretation of the functional data characterising the projection neurones of the cochlear nucleus (CN) is impossible without the unequivocal classification of these cell types at the end of the experiments. In this study, morphological criteria applicable for unambiguous identification of CN neurones have been sought. The neurones were labelled with rhodamine from incisions severing the projection pathways of the individual cell types, allowing their selective labelling and morphological characterisation. Confocal microscopy was employed for the investigation of the rhodamine-filled cells whose morphology was assessed after reconstructing the three-dimensional images of the cell bodies and proximal processes. The diameters of the somata and the number of processes originating from the cell bodies were also determined. In most of the cases, unambiguous identification of the bushy, octopus and Purkinje-like cells was relatively straightforward. On the other hand, precise classification of the pyramidal cells was often difficult, especially because giant cells could easily possess morphological features resembling pyramidal neurones. Occasionally, giant cells also mimicked the appearance of octopus neurones, which may be another important source of identification error, especially as these two cell types are often situated close to each other in the CN. It is concluded that morphological criteria defined in the present work may be effectively applied for the unambiguous identification of the projection neurones of the CN, even following functional measurements, when the correct cell classification is essential for the interpretation of the experimental data. Moreover, the present study also confirmed that Purkinje-like cells project to the cerebellum.

Introduction: Coronary sinus (CS) leads used for cardiac resynchronization have undergone development in the last years. However, dislocation rate remained high (5-9%). The aim of this study was to investigate the effectiveness and safety of stent implantation in a CS side vein to stabilize the left ventricular lead position after postoperative or intraoperative dislocation of the electrode., Methods and Results: Thirty-six patients (age: 64 +/- 9.7 years, 19 primary, 17 ischemic cardiomyopathy, NYHA class III: 30, IV: 6) were treated with stenting. The procedure was performed because of postoperative dislocation in seven patients, while dislocation was observed during the implantation in 29 cases. The electrode was repositioned into the desired position and a bare metal coronary stent was introduced via another guide wire. The stent was deposited at 5- to 15-mm proximal to the tip of the electrode with a pressure of 6 to 14 atmospheres. Control angiography showed no blood flow compromise in any of the side branches or in the coronary sinus. Control echocardiography did not show pericardial effusion due to stenting. During follow-up (11.5 +/- 5.5, 2-23 months), left ventricular pacing threshold increased from 2.2 to 5.6 V in one patient, but dislocation was not found by fluoroscopy. Clinically important pacing threshold increase was not detected in the other cases. Impedance measurements did not suggest lead insulation failure., Conclusion: Stent implantation to stabilize the left ventricular lead position seems to be a useful and safe procedure in the treatment of patients with complicated coronary sinus anatomy or lead instability.

Caliciviruses closely related to human norovirus and sapovirus were recently detected in domestic pigs, causing discussions about the animal reservoir and the potential for zoonotic transmission to humans. To detect porcine caliciviruses, 17 fecal samples collected on two swine farms in southwestern Hungary were tested by reverse transcription-polymerase chain reaction. Three (17.6%) samples were positive for caliciviruses. This study confirms the presence of caliciviruses, both porcine sapovirus (genus Sapovirus) and porcine norovirus (genus Norovirus), in domestic pigs in Hungary and provides additional information on the viral genetic diversity and relationship to viruses referred to as human caliciviruses.

Dobrava hantavirus (DOBV) infection was diagnosed in a previously healthy 46-y-old hunter suffering from severe haemorrhagic fever with renal syndrome (HFRS). Specific IgM antibodies against DOBV were identified by an immunofluorescence assay, while viral nucleic acid was detected by the molecular method, confirming the diagnosis. Our results reveal an existing risk of DOBV transmission to humans in Hungary.

Background: Group C rotaviruses are recognized enteric pathogens of humans and animals. Human group C rotaviruses have been associated with sporadic episodes and large outbreaks of gastroenteritis in children and adults but their epidemiology and ecology are still unexplored., Objectives: To collect epidemiological data on group C rotavirus infections among children with gastroenteritis in Hungary and perform molecular characterization on the identified strains., Study Design: Fecal samples were collected during the 2003 surveillance in Baranya County, Hungary. The presence of group C rotavirus RNA was investigated by polyacrylamide gel electrophoresis and by reverse transcription-nested polymerase chain reaction for the VP6 gene. The identified strains were further characterized by sequencing and phylogenetic analysis of the VP7, VP6, VP4, and NSP4 genes., Results: Three of 472 samples (0.6%) tested positive for group C rotavirus. Two samples were selected for molecular analysis. Strains BaC 6104/03 and BaC 11549/03 displayed an overall identity of >99.8% and 99.3% at the nucleotide and amino acid level, respectively. The VP7 of the strain BaC 6104/03 was most closely related (99.5% aa) to the Nigerian strain Jajeri, while the VP4s of strains BaC 6104/03 and BaC 11549/03 were more similar (98.1% aa) to strains Belem and 208, detected in Brazil and China, respectively., Conclusions: Based on this 1-year study, we conclude that group C rotaviruses are not of epidemiological relevance in the etiology of childhood acute gastroenteritis in Hungary. The low sequence divergence between the Hungarian strains suggested that a single group C rotavirus strain circulated in this period in the study area.

Aims: Routine procedures for monitoring viruses in water samples have not been drawn up for the water-microbiology screening panel. Enteric viruses, including astroviruses, are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. The aim of this study was to detect human astroviruses (HAstVs) from raw wastewater samples., Methods and Results: To obtain data on whether human astroviruses are shed in the environment, 35 raw sewage samples from 22 sewage plants in different regions of Baranya County, Hungary were tested for astrovirus using a polyethylene glycol method for concentration and a guanidinium thiocyanate-silica procedure for extraction of viral RNA. Reverse transcription-polymerase chain reaction (RT-PCR) with HAstV-specific primer pairs was used for amplification and the specificity of amplicons was confirmed by nucleotide sequencing and phylogenetic analysis. Among the 35 raw sewage samples, 15 (43%) contained HAstV and by sequence analysis, 10 genotype HAstV-1 and one genotype HAstV-2 were identified., Conclusions: The high detection rate of astroviruses we encountered in this study provide convincing evidence that HAstVs circulate at a relatively high frequency in the Hungarian population. No correlation between the standard indicators of faecal pollution and the presence of HAstVs was found., Significance and Impact of the Study: Our study is the first report on detection of HAstV in sewage in Hungary and suggests that HAstV might be potent indicators of viral pollution in environmental specimens.

Amplification of the kcnk9 gene and overexpression of the encoded channel protein (TASK-3) seems to be involved in carcinogenesis. In the present work, TASK-3 expression of melanoma cells has been studied. For the investigation of TASK-3-specific immunolabelling, a monoclonal antibody has been developed and applied along with two, commercially available polyclonal antibodies targeting different epitopes of the channel protein. Both primary and metastatic melanoma cells proved to be TASK-3 positive, showing prominent intracellular TASK-3-specific labelling; mostly concentrating around or in the proximity of the nuclei. The immunoreaction was associated with the nuclear envelope, and with the processes of the cells and it was also present in the cell surface membrane. Specificity of the immunolabelling was confirmed by Western blot and transfection experiments. As TASK-3 immunopositivity of benign melanocytes could also be demonstrated, the presence or absence of TASK-3 channels cannot differentiate between malignant and non-malignant melanocytic tumours.