Analysis of microRNA expression in brush cytology specimens improves the diagnosis of pancreatobiliary cancer.

BACKGROUND/OBJECTIVES: Malignant pancreatobiliary strictures are in many cases clinically indistinguishable and present a major problem to endoscopy specialists. Intraductal sampling procedures such as brush cytology are commonly used for diagnosis with a sensitivity that is low for a diagnostic test used in daily clinical practice. MicroRNA (miR) alterations detected in many cancers are disease-specific, which can be utilized in clinical applications. The aim of the present study was to analyze whether determination of miR expression levels in intraductal brush cytology specimens is a feasible approach to improve the diagnosis of pancreatobiliary cancer. METHODS: Brush cytology specimens have been collected during endoscopic retrograde cholangio-pancreatography (ERCP) and analyzed by routine cytology and ancillary miR assays. Total RNA was extracted using the miRNeasy Mini Kit and the expression of miRs frequently dysregulated in pancreatobiliary cancer (miR-16, miR-21, miR-196a, miR-221) were analyzed by quantitative real-time PCR using RNU6B as internal control. RESULTS: Routine cytology resulted in no false positive diagnoses, however, the combined sensitivity remained at 53.8%. Expression (ΔCt values) of miR-16 (p = 0.0039), miR-196a (p = 0.0003) and miR-221 (p = 0.0049) showed a clear statistical significance between malignant and benign pancreatobiliary specimens (n = 35). Malignancy could be detected combining routine cytology and the miR-196a single marker expression levels with a sensitivity of 84.6% (92.9% in biliary strictures) with no false positives. CONCLUSIONS: The results offer the first direct demonstration that microRNAs are readily detectable in brush cytology specimens obtained during ERCP, and have the potential to help the cytological diagnosis of pancreatobiliary malignancy.

Role of HLA-DRB1 and PTPN22 genes in susceptibility to juvenile idiopathic arthritis in Hungarian patients.

OBJECTIVE: Juvenile idiopathic arthritis (JIA) is a complex immune-mediated disease characterized by environmental influences along with several predisposing genes in the pathogenesis. The present study was undertaken to investigate the association of polymorphisms in two candidate genes for autoimmunity, human leukocyte antigen (HLA) DRB1 and protein tyrosine phosphatase N22 (PTPN22) with JIA in Hungarian patients. METHODS: A case-control study including 150 Hungarian JIA patients and 200 sex and ethnically matched healthy controls was conducted. Genotyping for HLA-DRB1 and PTPN22 C1858T single nucleotide polymorphism (SNP) (rs2476601) was carried out by group-specific PCR amplification and by real-time PCR allelic discrimination, respectively. RESULTS: In Hungarian patients JIA was associated with HLA-DRB1*01, DRB1*08, DRB1*13 (p=0.048, p=0.002, p=0.019, respectively) with marked differences between the disease subtypes classified according to the ILAR criteria. There was no association of the PTPN22 C1858T SNP with JIA (p=0.66). No correlation was found between the presence of this PTPN22 SNP and HLA-DRB1 alleles. CONCLUSIONS: Our results confirm that certain HLA-DRB1 alleles reported previously as susceptibility factors are strongly associated with JIA in a Hungarian population. However, C1858T polymorphism of PTPN22, another candidate gene of autoimmunity seems to be independent of JIA in Hungarian patients. Our data taken together with various findings in different populations suggest that associations related to PTPN22 seem to be more ethnicity-specific in contrast to the general and less population-dependent role of HLA-DRB1 in JIA.

Wide-range transcriptional modulating effect of ntrR under microaerobiosis in Sinorhizobium meliloti.

A mutation in the second gene in the ntrPR operon results in increased expression of nodulation (nod) and nitrogen fixation (nif) genes in Sinorhizobium meliloti. Since this pleiotropic effect is particularly pronounced in the presence of external combined nitrogen, a nitrogen regulatory function has been suggested for NtrR. To identify the complete set of protein-coding genes influenced by loss of ntrR function, microarray hybridizations were carried out to compare transcript levels in the wild type and mutant strains grown under aerobic and microaerobic conditions. Of the 6207 genes examined, representing the entire genome of S. meliloti, 7% exhibited altered expression: 4.5% of the genes are affected under oxic, 2.5% under microoxic conditions. 0.4% of all the genes are affected under both oxygen concentrations. A microoxic environment is required for the induction of genes related to symbiotic functions but results in the down-regulation of other (e.g. metabolic) functions. When the alterations in transcription levels at low oxygen concentration in the mutant strain were compared to those of the wild type, a modulating effect of the ntrR mutation was observed. For example, symbiotic nif/fix genes were induced in both strains, but the level of induction was higher in the ntrR mutant. In contrast, genes related to transcription/translation functions were down-regulated in both strains, and the effect was greater in the wild-type strain than in the ntrR mutant. A relatively wide range of functions was affected by this modulating influence, suggesting that ntrR is not a nitrogen regulatory gene. Since genes encoding various unrelated functions were affected, we propose that NtrR may either interfere with general regulatory mechanisms, such as phosphorylation/dephosphorylation, or may influence RNA stability.

Effects of intraamniotically administered thyroxin on acceleration of fetal pulmonary maturity in preeclamptic toxemia.

In 18 cases of serious and fairly serious toxemic pregnancy, the authors gave 500 micrograms L-Thyroxin "Henning" intraamniotically after the laboratory evaluation demonstrated negative pulmonary maturity from an amniotic fluid sample. Maternal and fetal complications due to the drug could not be observed. Despite a 39% premature incidence, IRDS and hyaline membrane disease were not observed. The administered T4 caused a positive change in the direction of the L/S ratio and the Clements test. The other amniotic fluid parameters did not change, except the T4 level. The extremely high T4 level obtained 48 hours later gradually became normal several days later. The T4 values obtained from the blood of the mother and the newborn on the 5th postnatal day were normal. In toxemic cases where the induction of labor is vital and there is a risk of IRDS and steroid application in contraindicated, intraamniotic thyroxin is recommended as prophylaxis for IRDS.

Isolation of aerobic bacteria from the placenta.

Cultures for aerobic bacteria were prepared from 353 placentas. Specimens were taken from the chorion after removing the amnion. The specimens were immersed into Stuart transport medium. Microscopic examination of the placenta and cultures from the throat and ear of newborns were also done. The rate of positive bacterial cultures was 16%. Chorioamnionitis was found in 15%. The proportion of chorioamnionitis caused by aerobic bacteria was 44%. The rate of positive bacterial cultures from the placenta in the group of newborns with clinical signs of intrauterine infection was 63%. Bacteria can be present on the chorionic plate without any histological evidence of chorioamnionitis. Bacteriological examination of the placenta is therefore mandatory when amniotic fluid infection is suspected.

Effect of steroid prophylaxis and of elective caesarean section on maternal and fetal cortisol values.

The cortisol level was studied during and after delivery in the mothers and their newborns after normal deliveries, elective Caesarean sections, and steroid pre-treated vaginal deliveries. The suppressive effect of Caesarean section, and steroid pre-treated vaginal deliveries. The suppressive effect of exogenous steroid was observed in both the mother and the newborn but its extent was not significant and its duration transitory; in 48 hours it disappeared. In the newborns delivered by elective Caesarean section, an extreme increase in the cortisol level was observed. The cortisol level in the amniotic fluid was not suitable for evaluating the function of the hypophysis-adrenal axis, especially not after steroid prophylaxis.