Attila Juhász

Cerebrovascular reactivity in hypertensive patients: A transcranial Doppler study

We studied the usefulness of transcranial Doppler sonography for assessing changes in vasoreactivity in patients with hypertension and the hemodynamic consequences of hypertension.

Reference genes for quantitative real time PCR in UVB irradiated keratinocytes.

Real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a sensitive and highly reproducible method often used for determining mRNA levels. To enable proper comparison of gene expression genes expressed at stabile levels within the cells in the studied experimental system need to be identified and used as reference. Ultraviolet B (UVB) radiation is an exogenous carcinogenic stimulus in keratinocytes, and UVB elicited changes have extensively been studied by qRT-PCR, yet a comparison of commonly used reference genes in UVB treatment is lacking. To find the best genes for compensating slight inter-sample variations in keratinocytes in UVB experiments and to understand the potential effects of improper reference gene (RG) selection we have analyzed the mRNA expression of 10 housekeeping genes in neonatal human epidermal keratinocytes (NHEK) after UVB treatment. The biological effect of the used UVB light source was validated by trypane blue exclusion, MTT and comet assays. 20-40mJ/cm(2) dose was chosen for the experiments. The stability of the 10 RGs was assessed by the GeNorm and Normfinder software tools. Regardless of their slightly different algorithm the programs found succinate dehydrogenase complex subunit A (SDHA) to be the best individual RG and SDHA and phosphoglycerate kinase-1 (PGK1) as the most suitable combination. Analysis of the expression of tumor necrosis factor alpha (TNFalpha) and vascular endothelial growth factor (VEGF) found that while the perception of changes in TNF-alpha, a gene undergoing marked upregulation after UVB irradiation is independent of the used RG, changes seen in the more modestly upregulated VEGF are greatly effected by reference gene selection. These findings highlight the importance of reference gene selection in UVB irradiation experiments, and provide evidence that using SDHA or the combination of SDHA and PGK1 as standards could be a reliable method for normalizing qRT-PCR results in keratinocytes after UVB treatment.

Expression of invasion markers CD44v6/v3, NM23 and MMP2 in laryngeal and hypopharyngeal carcinoma

Twelve laryngeal squamous cell carcinoma cases (7 laryngeal and 5 hypopharyngeal cancer; 15 samples) were analysed by immunohistochemistry for the expression of invasion markers CD44v6/v3, NM23 and matrix metalloproteinase, MMP2. The laryngeal epithelium showed CD44v6+v3+NM23- /MMP2- phenotype. When tumors were grouped into TNM categories the phenotype of the T2 and T3 tumors was similar, characterised by decreased CD44v3+ and lack of MMP2 expressions. Meanwhile the NM23 expression was more frequent in T3 tumors. In T4 stage the frequency of NM23 and MMP2 positive cases increased (5/6 and 4/6, respectively) but there was no correlation with the appearence of lymph node metastasis. Comparison of the phenotype of laryngeal and hypopharyngeal tumors, irrespective of the TNM stages, revealed characteristic differences: T2 stage laryngeal tumors showed decreased CD44v3 and occasional NM23 and MMP2 positivity, while in T3 stage these tumors were characterised by increased frequency of NM23 positivity. The phenotype of the hypopharyngeal tumors was significantly different with a high frequency of MMP2 positive cases (5/6) and NM23+1ow CD44v3+ phenotype. The sharp differences in the phenotypes of laryngeal and hypopharyngeal carcinomas were connected to the differences in their invasive capacity unlike to the size of the tumors, since the T4 stage hypopharyngeal tumors had a significantly smaller size than laryngeal ones, even at lower stages.

Elevated level of tenascin and matrix metalloproteinase 9 correlates with the bone destruction capacity of cholesteatomas

Objectives: Bone destruction is a key step in the progression of cholesteatomas. Some of the lesions can grow without affecting the surrounding anatomic structures, whereas others can cause severe bone destruction despite their limited size. This study aims to identify factors that could play important role during the invasion of the disease. Methods: Cholesteatoma tissue samples were examined immunohistochemically. Tissue samples were arranged on the basis of bone destruction (destructive cholesteatomas [DC] and nondestructive cholesteatomas [NDC]). Double-immunofluorescent labeling was performed to detect simultaneously 1) tenascin (TN) and cytokeratin; 2) matrix metalloproteinase 9 (MMP-9) and TN; 3) TN and Ki-67. An in situ apoptosis detection kit was used to detect apoptotic cells. External auditory canal skin samples were used as control. Results: 1) In DCs, more widespread stromal TN labeling was seen compared with NDCs or external auditory canal skin samples. 2) More enhanced MMP-9 staining was detected in DCs compared with NDCs. 3) The proportion of Ki-67-positive cells in DC samples was significantly higher than in NDCs. 4) The percentage of apoptotic cells was higher in NDC than in DC samples. Conclusion: Our present study demonstrates that levels of TN, MMP-9, and proliferative activity are increased in cholesteatomas. It has also been shown that increased levels of TN, MMP-9-positive cells, and proliferative activity of the lesions, as well as decreased levels of apoptosis, can be linked to more aggressive clinical behavior of cholesteatomas. Our findings also indicate that TN and MMP-9 can be key molecules of bone destruction during cholesteatoma progression.

Chromosomal imbalances in laryngeal and hypopharyngeal cancers detected by comparative genomic hybridization

The prognostic divergence of laryngeal and hypopharyngeal carcinomas is well known. Hypopharyngeal tumors are characterized by frequent metastasis formation and local recurrence, which is the source of the unfavorable prognosis of this subtype. The aim of this study was to define chromosomal alterations associated with the aggressive behavior of hypopharyngeal tumors.

Characteristic distribution patterns of tenascin in laryngeal and hypopharyngeal cancers

Objectives: Progression of malignant neoplasias is accompanied by alteration of the extracellular matrix (ECM) composition. Tenascin is known as a member of the adhesion‐modulating family of ECM macromolecules; thus its expression and distribution may have significant influence on tumor cell proliferation and invasiveness.

Development of a Deprivation Index and its relation to premature mortality due to diseases of the circulatory system in Hungary, 1998-2004.

An association between health and socio-economic status is well known. Based on international and national studies, the aims of this study were to develop a multi-dimensional index at the municipality level, to provide information about socio-economic deprivation in Hungary and to investigate the association between socio-economic status and the spatial distribution of premature mortality due to diseases of the circulatory system. Seven municipality level socio-economic indicators were used from the National Information System of Spatial Development (income, low qualification, unemployment, one-parent families, large families, density of housing and car ownership). After normalisation and standardisation, indicator weights were evaluated using factor analysis. A risk analysis study was conducted using the Rapid Inquiry Facility software to evaluate the association between deprivation and the spatial distribution of premature mortality due to diseases of the circulatory system for the years 1998-2004. Areas of significantly high deprivation were identified in the northeastern, eastern and southwestern parts of Hungary. A statistically significant association was found between premature cardiovascular mortality and deprivation status in both genders. The Deprivation Index is the first composite index at the municipality level in Hungary and includes key factors that affect socio-economic status. The identified association highlighted the fact that inequalities in socio-economic status may reflect the spatial distribution of health status in a population. The results can be used to inform prevention strategies and help plan local health promotion programs aimed at reducing health inequalities.

Relationship of Serum Resistin Level to Traits of Metabolic Syndrome and Serum Paraoxonase 1 Activity in a Population with a Broad Range of Body Mass Index

UNLABELLED The relationship between resistin, one of the adipokines, and metabolic syndrome is not fully elucidated. Altered activity of the HDL-associated antioxidant enzyme paraoxonase 1 (PON1) that participates in the antioxidant defense mechanisms of HDL may have an important role in the obesity-related accelerated atherosclerosis. Inverse associations of PON1 with obesity and serum levels of leptin have been demonstrated. Our aim was to investigate the association of serum levels of resistin with (i) PON1 activity, and (ii) parameters of metabolic syndrome, including some that are additional for research. A total of 74 Caucasian subjects were recruited into the study and divided into 3 age and sex-matched groups. Group 1, 25 non-diabetic overweight/obese subjects with BMI of 28-39.9 kg/m (2); group 2, 25 non-diabetic obese patients with BMI >or=40 kg/m (2); and the control group 3, 24 healthy, normal-weight control subjects. Serum levels of resistin were correlated negatively with BMI (r=-0.27, P<0.05), waist circumference (r=-0.28, P<0.05), serum levels of leptin (r=-0.28, P<0.05), non-esterified fatty acids (NEFA) (r=-0.23, P<0.05), and HbA (1C) (r=-0.26, P<0.05), systolic BP (r=-0.28, P<0.05), and lipid peroxidation (measured by TBARS) (r=-0.40, P<0.01), and correlated positively with PON1 (r=0.24, P<0.05). No association was detected between the serum concentrations of resistin and the following investigated parameters: diastolic BP, levels of uric acid, glucose, insulin, or insulin resistance (measured by homeostasis model assessment, HOMA-IR), triglyceride, total cholesterol, LDL-C, and HDL-C. During multiple regression analyses BMI and TBARS were independent predictors of PON1, while age, gender, blood pressure, HOMA-IR, LDL-C, HDL-C, and resistin were not. CONCLUSIONS Among the study subjects, serum levels of resistin showed a positive, although not independent correlation with serum PON1, and a negative correlation with numerous parameters of the metabolic syndrome (i.e. adiposity, blood pressure, levels of leptin, free fatty acid, glycosylated hemoglobin, and lipid peroxidation). BMI and TBARS are independent predictors of PON1 activity.

Cardiovascular effects of neuropeptide FF antagonists

The neuropeptide FF (NPFF) antagonist desaminotyrosyl-Phe-Leu-Phe-Gln-Pro-Gln-Arg-NH2 dose-dependently reversed NPFF-induced elevation of blood pressure in anesthetized rats after intravenous injection without causing a significant change of blood pressure and heart rate by itself. However, another antagonist dansyl-Pro-Gln-Arg-NH2 produced a significant drop of the mean arterial pressure only at a large dose (10 micromol/kg body weight), but reversal of the NPFF-induced hypertension was modest. Consequently and contrary to the conclusions of a previous study, NPFF antagonists cannot be identified simply by measuring the changes in the hemodynamic parameters upon the injection of the compounds alone and without a subsequent NPFF challenge.

Persisting TT virus (TTV) genogroup 1 variants in renal transplant recipients

Summary. TT virus (TTV) genogroup 1 infection has an increased prevalence in solid organ transplant recipients. In this study, the presence of TTV in renal transplant recipients was examined by two PCR methods, one capable of detecting most TTV genotypes (UTR-PCR), the other specific to genogroup 1 (N22-PCR). The N22-PCR detected TTV in 57% (53/92) of the renal transplant patients and in 20% (13/66) of the healthy individuals, while the prevalence of TTV with the UTR-PCR was above 90% in both the control and the patient groups. The N22-PCR was used in longitudinal studies of 31 renal transplant recipients, these PCR products were sequenced and aligned. TTV status was not associated with the patients’ age at transplantation, male to female ratio and the time lag between kidney transplantation and the TTV test. During the follow-up consistent TTV status was found in 26 patients, while two initially TTV positive patients converted to negative and three initially negative patients converted to positive. The TTV variants varied among the tested patients, but were the same in the consecutive samples of each patient, indicating that TTV infection was persistent in renal transplant recipients and novel infection occured rarely in the post-transplant period.