Investigation of the Effects of PACAP on the Composition of Tear and Endolymph Proteins

Pituitary adenylate cyclase activating polypeptide (PACAP) is widely distributed in ocular tissues, including the lacrimal gland. PACAP has been shown to influence the activity of several exocrine glands, but its effects on the composition of the tear film are not known yet. Similarly, the presence of PACAP has already been shown in the inner ear, but it is not known whether PACAP influences the composition of the endolymph. The aim of the present study was to investigate whether systemic injection of PACAP has any modulatory effects on the protein composition of the tear film and endolymph using chip electrophoresis and mass spectrometry analysis. Tear and endolymph samples were collected from rats and chickens, respectively, at various time points after systemic injection of PACAP. Fluid samples were further processed for chip electrophoretic studies. No difference was found in the protein composition of the endolymph between control and PACAP-treated animals. In contrast, tear samples showed a marked difference after PACAP treatment. Proteins in the molecular range 50-70 kDa, which showed a different chip electropherogram profile in every PACAP-treated sample, were further analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PACAP treatment induced a repression in certain keratins, while others were induced after PACAP injection. Furthermore, PACAP treatment decreased aldehyde dehydrogenase expression. The present study provides a base for further studies on the in vivo effects of PACAP on the composition of tear film. These investigations may have important clinical relevance because of the noninvasive sample collection, the correlation between tear proteins and ocular diseases, and the possible presence of biomarkers for both ophthalmological and systemic pathological conditions.     

Microvascular reactivity in lean, overweight, and obese hypertensive adolescents

The microvascular responses to endothelium-dependent vasodilators (e.g., acetylcholine), endothelium-independent vasodilators (e.g., sodium nitroprusside), and to local heating were studied (for the first time) in adolescents with essential hypertension, grouped according to their body mass index. The forearm microvascular reactivities of thirty-three hypertensive adolescents (ten lean, 13 overweight, and ten obese) and 19 healthy controls were assessed by means of laser Doppler flowmetry. Blood levels of enzymatic and nonenzymatic antioxidants and malondialdehyde were determined. The perfusion increments in response to acetylcholine iontophoresis were not significantly attenuated in the patient groups as compared with the controls. Sodium nitroprusside (SNP) iontophoresis resulted in significantly smaller perfusion increments in the lean and obese hypertensives than in the controls (both p?<?0.05). Similar responses to local heating (44°C) performed after either acetylcholine or SNP iontophoresis were observed at the respective measurement sites. As compared with the controls, we found elevated ratios of the whole blood oxidized and reduced glutathione in all the patient groups (all p?<?0.001), increased erythrocyte catalase activities in the overweight hypertensives (p?<?0.05), and decreased ratios of the plasma alpha-tocopherol and triglycerides in the obese hypertensive group (p?<?0.05). Conclusion: The endothelium-dependent microvascular reactivity was not significantly attenuated in the hypertensive adolescents in contrast with the impaired endothelium-independent vasorelaxation in the lean and obese hypertensives.     

Drastic changes in serotypes of carried pneumococci due to an increased vaccination rate in Hungary

Introduction of the conjugate pneumococcal vaccine into the voluntary childhood vaccine program in Hungary in April 2009 resulted in a sharp increase of the vaccination rate. However, changes in serotypes as a consequence of vaccination should be considered. Aims: The aim of the authors was to compare pneumococci isolated from children with high-level and low-level vaccination rates. Methods: Nasal specimens from 854 children attending 20 nurseries at various locations in Hungary have been collected since 2009. The serotypes, antibiotic susceptibility and genetic relatedness of the isolated pneumococci were determined. Results: 324 strains were isolated, and the carriage rate was 37.94%. The strains were sensitive to most antibiotics, except for macrolides. A definite suppression of vaccine types was detected during these 3 years, from the initial 78.85% to 35.30%. Conclusions: The authors conclude that the results reflect the efficacy of the vaccine, which underlines the need for the inclusion of pneumococcal vaccine into the list of obligatory vaccines. Orv. Hetil., 2012, 153, 1031-1034.     

Second window of protection following myocardial preconditioning: an essential role for PI3 kinase and p70S6 kinase

Ischaemic preconditioning (IPC) protects the heart against myocardial infarction acutely as well as several hours later (e.g. 24-48 h). The mechanism of the profound cardioprotection is not completely explored. We hypothesized that PI3K/PDK1/Akt/mTOR/p70S6K-mediated pro-survival pathway is involved in delayed cardioprotection induced by IPC. Under Hypnorm-Diazepam anaesthesia, male New Zealand White rabbits were either sham-operated (SC) or preconditioned by four cycles of 5-min ischaemia and 10-min reperfusion on day 1. Twenty-four hours after recovery, the animals were anaesthetized with sodium pentobarbitone and subjected to 30-min ischaemia followed by 180-min reperfusion. Wortmannin (0.6 mg/kg, i.v.), an irreversible PI3 kinase (PI3K) inhibitor, rapamycin (0.25 mg/kg, i.v.), which prevents the phosphorylation of p70S6 kinase (p70S6K), or DMSO (control vehicle) was given 15 min prior to IPC. IPC significantly reduced infarct size compared to the control group (SC) (31.9 +/- 5.8% (n = 7) vs. 54.9 +/- 2.9% (n = 6), P < 0.05). Wortmannin and rapamycin alone had no effect on infarct size (56.3 +/- 1.6% (n = 6) and 54.7 +/- 3.8% (n = 6), respectively). However, when wortmannin or rapamycin were given prior to IPC the protection was completely abolished (49.9 +/- 2.8% (n = 6), 45.1 +/- 4.6% (n = 7), P < 0.05 vs. IPC). Western blot analysis showed that wortmannin, at a dose of 0.6 mg/kg, and rapamycin, at a dose of 0.25 mg/kg, were sufficient to prevent phosphorylation of Akt and p70S6K, respectively, when the inhibitors were given prior to IPC. We conclude that PI3K/PDK1/Akt/mTOR/p70S6K-signalling pathway plays an essential role in the development of the cardioprotection against infarction in rabbits.     

Hospital and long-term prognosis of patients with non-ST-segment elevation myocardial infarction

The authors analyze some clinical data, hospital and long-term prognosis of 139 consecutive patients with non-ST-segment elevation myocardial infarction. A three-quarter-year long follow-up of patients was performed, and data were recorded by use of postal questionnaire. The follow-up was complete in 98% of the patients. The mean age was 78.6 and 71.4 years in the case of female and male patients, respectively. High percent of patients had comorbidity (diabetes mellitus, hypertension and previous ischemic heart disease). Coronary angiography was performed in 30 patients (22%) and revascularization in 29 of them. Hospital mortality was found in 15% and during follow-up 17% of the patients died. The patients who died during hospital stay and during the follow-up period were significantly older than patients who stayed alive. Some echocardiographic parameters were also found prognostically important: ejection fraction, end systolic diameter, segmental wall motion abnormality and severity of mitral regurgitation were found significantly different in patients alive and who died. The authors investigated the hospital discharge medications according to guideline recommendations. The authors emphasize that this quality indicator was much better in this patient population than the earlier published data. By the end of follow-up, similar percent of patients used these drugs as at the time of hospital discharge.     

Performance of a new HPV and biomarker assay in the management of hrHPV positive women: Subanalysis of the ongoing multicenter TRACE clinical trial (n > 6,000) to evaluate POU4F3 methylation as a potential biomarker of cervical precancer and cancer

The ongoing Triage and Risk Assessment of Cervical Precancer by Epigenetic Biomarker (TRACE) prospective, multicenter study aimed to provide a clinical evaluation of the CONFIDENCE? assay, which comprises a human papillomavirus (HPV) DNA and a human epigenetic biomarker test. Between 2013 and 2015 over 6,000 women aged 18 or older were recruited in Hungary. Liquid‐based cytology (LBC), high‐risk HPV (hrHPV) DNA detection and single target host gene methylation test of the promoter sequence of the POU4F3 gene by quantitative methylation‐specific polymerase chain reaction (PCR) were performed from the same liquid‐based cytology sample. The current analysis is focused on the baseline cross‐sectional clinical results of 5,384 LBC samples collected from subjects aged 25 years or older. The performance of the CONFIDENCE HPV? test was found to be comparable to the cobas® HPV test with good agreement. When applying the CONFIDENCE Marker? test alone in hrHPV positives, it showed significantly higher sensitivity with matching specificity compared to LBC‐based triage. For CIN3+ histological endpoint in the age group of 25–65 and 30–65, the methylation test of POU4F3 achieved relative sensitivities of 1.74 (95% CI: 1.25–2.33) and 1.64 (95% CI: 1.08–2.27), respectively, after verification bias adjustment. On the basis of our findings, POU4F3 methylation as a triage test of hrHPV positives appears to be a noteworthy method. We can reasonably assume that its quantitative nature offers the potential for a more objective and discriminative risk assessment tool in the prevention and diagnostics of high‐grade cervical intraepithelial neoplasia (CIN) lesions and cervical cancer.     

Comparison of different lymph node staging systems in prognosis of gastric cancer: a bi-institutional study from Hungary

Objective:The Union for International Cancer Control (UICC) Node (N) classification is the most common used staging method for the prognosis of gastric cancer.It demands adequate,at least 16 lymph nodes (LNs) to be dissected;therefore different staging systems were invented.Methods:Between March 2005 and March 2010,164 patients were evaluated at the Department of General Surgery in the Ken(e)y Gyula Hospital and at the Department of General,Thoracic and Vascular Surgery in the Kaposi Mór Hospital.The 6th,7th and 8th UICC N-staging systems,the number of examined LNs,the number of harvested negative LNs,the metastatic lymph node ratio (MLR) and the log odds of positive LNs (LODDS) were determined to measure their 5-year survival rates and to compare them to each other.Results:The overall 5-year survival rate for all patents was 55.5％ with a median overall survival time of 102 months.The tumor stage,gender,UICC N-stages,MLR and the LODDS were significant prognostic factors for the 5-year survival with univariate analysis.The 6th UICC N-stage did not follow the adequate risk in comparing N2 vs.N0 and N3 vs.N0 with multivariate investigation.Comparison of performances of the residual N classifications proved that the LODDS system was first in the prediction of prognosis during the evaluation of all patents and in cases with less than 16 harvested LNs.The MLR gave the best prognostic prediction when adequate (more than or equal to 16) lymphadenectomy was performed.Conclusions:We suggest the application of LODDS system routinely in western patients and the usage of MLR classification in cases with extended lymphadenectomy.     

The Value of a Novel Panel of Cervical Cancer Biomarkers for Triage of HPV Positive Patients and for Detecting Disease Progression

In the era of primary vaccination against HPV and at the beginning of the low prevalence of cervical lesions, introduction of screening methods that can distinguish between low- and high-grade lesions is necessary in order to maintain the positive predictive value of screening. This case-control study included 562 women who attended cervical screening or were referred for colposcopy and 140 disease free controls, confirmed by histology and/or cytology. The cases were stratified by age. Using routine exfoliated liquid based cytological samples RT-PCR measurements of biomarker genes, high-risk HPV testing and liquid based cytology were performed and used to evaluate different testing protocols including sets of genes/tests with different test cut-offs for the diagnostic panels. Three new panels of cellular biomarkers for improved triage of hrHPV positive women (diagnostic panel) and for prognostic assessment of CIN lesions were proposed. The diagnostic panel (PIK3AP1, TP63 and DSG3) has the potential to distinguish cytologically normal hrHPV+ women from hrHPV+ women with CIN2+. The prognostic gene panels (KRT78, MUC5AC, BPIFB1 and CXCL13, TP63, DSG3) have the ability to differentiate hrHPV+ CIN1 and carcinoma cases. The diagnostic triage panel showed good likelihood ratios for all age groups. The panel showed age-unrelated performance and even better diagnostic value under age 30, a unique feature among the established cervical triage tests. The prognostic gene-panels demonstrated good discriminatory power and oncogenic, anti-oncogenic grouping of genes. The study highlights the potential for the gene expression panels to be used for diagnostic triage and lesion prognostics in cervical cancer screening.     

mRNA and microRNA expression patterns in adrenocortical cancer

Adrenocortical cancer is a rare tumor and its prognosis is poor. Although numerous tumor-associated genetic and signal transduction alterations have been described to date, its pathogenesis is still unclear. Hybridization-based DNA microarray approaches may reveal significant gene expression alterations and may thus contribute to a better understanding of tumorigenesis and may identify molecular markers applicable for the distinction of benign and malignant lesions. Beside gene expression patterns, studies on microRNAs seem to be useful, as well. Novel therapeutical targets might be established by these approaches. In this review, the authors attempt to summarize the main findings of mRNA and microRNA expression microarray studies performed to date in adrenocortical cancer including a recent meta-analysis of gene expression data and present novel pathogenic pathways.