Phenotype of dendritic cells generated in the presence of non-small cell lung cancer antigens - preliminary report.

Therapeutic outcomes of definitively treated non-small-cell lung cancer (NSCLC) are unacceptably poor. It has been proposed that the manipulation of dendritic cells (DCs) as a "natural" vaccine adjuvant may prove to be a particularly effective way to stimulate antitumor immunity. Presently, there is no standardized methodology for preparing vaccines and many questions concerning the optimal source and type of antigens as well as maturation state and activity of DCs are still unsolved. The study population comprised of ten patients with histologically confirmed NSCLC (mean age: 67.63 +/- 6.15 years). Resected small tumor pieces were placed in tissue culture dishes containing different growth factors in order to obtain pure cancer cells. Seven days after the operation, the PBMC were collected and monocytes were purified by the adherence to culture dishes. Monocytes were cultured in RPMI 1640 medium supplemented with 10% of autologous plasma in the presence of rhIL-4 and rhGM-CSF to generate immature autologous (DCs). TNF-alpha with or without tumor cells' lysate were added to maturation of DCs. After 7 days of culture, DCs were harvested and the expression of CD1a, CD83, CD80, CD86 and HLA-DR antigens were analyzed by flow cytometry. We discovered higher (p=0.07) percentage of semimature DCs in tumor cell lysate culture in comparison with TNF-alpha culture (21.22 +/- 16.82% versus 11.27 +/- 11.64%). The expression of co-stimulatory and maturation markers (CD86, CD83 and HLA-DR) was higher on DCs from the culture with tumor cell lysate compared with TNF-alpha culture as a control. Specimen of NSCLC's culture prepared in this way could generate differences in DCs phenotype, which may have an influence on the therapeutic and protective antitumor immunity of the vaccine. Our research seems to be the next step in the development of DC-based vaccine. We are going to continue the investigation to start the preparation of a pattern of immunological vaccine against lung cancer

Ocena równowagi limfocytów Th1/Th2 oraz ekspresji receptorów dla lipopolisacharydu u chorych na astmę

Introduction: An increase in the number of asthma patients which has recently been observed depends on their place of residence and their occupation. This suggests that both external factors and genetic predispositions affect the development of the disease. The contact with bacterial lypopolysaccharide (LPS) may suppress the development of asthma among rural inhabitants. The mechanism of LPS effect most probably consists in the activation of macrophages and granulocytes by TLR4 and CD14 receptors for the production of cytokines, which affect Th1/Th2 balance. The objective of the study was the evaluation of CD14 and TLR4 expression on mononuclear cells and the analysis of Th1/Th2 balance in peripheral blood among asthma patients. Material and methods: The study group covered 22 patients with bronchial asthma (mean age 45 ± 15), and was conducted by the method of flow cytometry with the use of fluorochrome-labelled monoclonal antibodies. CD14 and TLR expression was assessed in peripheral blood monocytes. Th1/Th2 balance was determined by the measurement of intracellular IL-2, IFN-γ, IL-4 and IL-10 expression in T-helper cells after culture with the stimulation of cytokine production. Results: A negative correlation was noted between TLR4 expression and the percentage of Th2 lymphocytes, while a positive correlation was observed between expression of TLR4 and percentage of Th1 cells. No relationship was found between CD14 expression on monocytes and the percentage of Th1 and Th2 lymphocytes. Conclusions: An increased percentage of lymphocytes with TLR4 expression is associated with the change in Th1/Th2 balance in favour of Th1 lymphocytes in asthma patients.Wstęp: Obserwowany obecnie wzrost zachorowań na astmę jest zależny od miejsca zamieszkania chorych i wykonywanej przez nich pracy. Sugeruje to, że na występowanie choroby mają wpływ zarówno czynniki zewnętrzne, jak również predyspozycje genetyczne. Kontakt z lipopolisacharydem (LPS) bakteryjnym może hamować rozwój astmy u osób z regionów rolniczych. Prawdopodobny mechanizm działania LPS polega na aktywowaniu makrofagów i granulocytów przez receptory TLR4 i CD14 do wytwarzania cytokin, mających wpływ na równowagę Th1/Th2. Celem pracy była ocena ekspresji CD14 oraz TLR4 na komórkach jednojądrzastych oraz analiza równowagi Th1/Th2 we krwi obwodowej u chorych na astmę. Materiał i metody: Grupę badaną stanowiło 22 chorych (wiek średni: 45 ± 15 lat) na astmę oskrzelową. Badanie wykonano metodą cytometrii przepływowej za pomocą przeciwciał monoklonalnych znakowanych fluorochromami. Ekspresję CD14 i TLR4 oceniono na wyizolowanych z krwi obwodowej komórkach jednojądrzastych. Równowagę Th1/Th2 określono przez wykonanie pomiaru wewnątrzkomórkowej ekspresji IL-2, IFN-γ, IL-4 i IL-10 w limfocytach T pomocniczych, w hodowlach prowadzonych ze stymulatorami wytwarzania cytokin. Wyniki: Odsetek limfocytów z ekspresją TLR4 korelował istotnie ujemnie z odsetkiem limfocytów Th2 i znamiennie dodatnio z odsetkiem limfocytów Th1. Nie stwierdzono zależności między ekspresją CD14 na monacytach a odsetkami limfocytów Th1 i Th2. Wnioski: Zwiększony odsetek limfocytów z ekspresją TLR4 ma związek z przesunięciem równowagi Th1/Th2 na korzyść limfocytów Th1 u pacjentów z astmą

How does the estimated phase of menstrual cycle or menopause influence the prevalence of vasovagal syncope induced by head-up tilt test

Background: The purpose of this study was to evaluate the prevalence of syncope induced by head-up tilt test (HUTT) and the type of positive vasovagal response to the orthostatic stress in a relationship to the estimated phase of menstrual cycle (follicular phase, luteal phase) or menopause. Methods: We observed a cohort of 500 women at age 13–89 years (median of age 37.5), referred to HUTT. Phase of the menstrual cycle was determined on the basis of the usual length of menstrual cycle and the day of the cycle at the time of the study. We assumed that the length of the luteal phase is constant and it is 14 days. Results: In premenopausal patients, the occurrence of the mixed and cardioinhibitory response was significantly more frequent in comparison to postmenopausal women (48.8 vs. 35.1% and 19.7 vs. 12.4%, respectively; p < 0.00001), while the occurrence of the vasodepressive one was significantly less frequent (7.3% vs. 28.6%; p < 0.00001) in that group of patients. Women in follicular phase presented vasodepressive response during HUTT more often than woman in the luteal phase (10.0% vs. 4.1%, p < 0.00001). Conclusions: Among women referred for HUTT, the prevalence of the vasovagal syndrome did not differ between those that were pre- and post-menopausal. Higher incidence of vasodepressive reaction in postmenopausal women was observed. Among the premenopausal women, the vasodepressive type of vasovagal syndrome occurred more often in follicular then in luteal phase

Molekularne terapie celowane w niedrobnokomórkowym raku płuca

Terapie celowane działają w sposób wybiórczy na białka sygnałowe komórek nowotworowych, powodując upośledzenie ich funkcji życiowych. W niedrobnokomórkowym raku płuca (NDRP) zastosowanie znalazły inhibitory kinaz tyrozynowych związanych z receptorem dla naskórkowego czynnika wzrostu (EGFR): erlotynib i gefitynib. Leki te są stosowane w zaawansowanym raku płuca jako terapia drugiego lub trzeciego rzutu po niepowodzeniu pierwszorzutowej chemioterapii. Cetuximab jest przeciwciałem monoklonalnym skierowanym przeciwko zewnątrzkomórkowej domenie EGFR. Natomiast bewacyzumab jest przeciwciałem blokującym działanie czynnika wzrostu śródbłonka naczyń (VEGF), powodując upośledzenie ukrwienia tkanki nowotworowej. Przeciwciała mogą być stosowane jako uzupełnienie chemioterapii oraz po jej zakończeniu. Mediana czasu życia chorych na zaawansowaną postać NDRP w wyniku stosowania bewacyzumabu wyniosła po raz pierwszy w historii leczenia tego nowotworu ponad 12 miesięcy. Istnieją duże różnice w skuteczności omawianych leków w zależności od genetycznych właściwości komórek nowotworowych. Obecnie najważniejszymi badaniami molekularnymi użytecznymi w kwalifikacji chorych do terapii celowanych są: badanie immunohistochemiczne w celu wykrycia ekspresji EGFR, fluorescencyjna hybrydyzacja in situ, za pomocą której można wykryć amplifikację genu dla EGFR, oraz metody genetyczne zmierzające do określenia występowania mutacji EGFR i K-Ras

Common variant p.D19H of the hepatobiliary sterol transporter ABCG8 increases the risk of gallstones in children

Introduction Gallstones are increasingly common in children. Genetic analyses of adult cohorts demonstrated that the sterol transporter ABCG8 p.D19H and Gilbert UGT1A1*28 variants enhance the odds of developing gallstones. The genetic background of common lithiasis in children remains unknown. Methods Overall, 214 children with gallstone disease (1 month–17 years, 107 boys) were inclueded. The control cohorts comprised 214 children (age 6–17 years, 115 boys) and 172 adults (age 40–92 years, 70 men) without gallstones. The ABCG8 p.D19H and UGT1A1*28 polymorphisms as well as ABCB4 (c.504C>T rs1202283, c.711A>T rs2109505) and NPC1L1 variants (p.V1296V rs217434, c.−18C>A rs41279633) were genotyped using TaqMan assays. Serum concentrations of plant sterols and cholesterol precursors were measured by gas chromatography/mass spectrometry. Results The ABCG8 risk allele was associated with an increased risk of stones (OR = 1.82, p = .03). Children carrying the p.19H allele presented with lower serum concentrations of surrogate markers of intestinal cholesterol absorption and decreased ratios of phytosterols to the cholesterol precursor desmosterol. Carriers of the common NPC1L1 rs217434 allele had an increased gallstone risk compared with stone-free adults (OR 1.90, p < .01). This variant also affected the ratio of phytosterols to cholesterol precursors (p = .03). Other tested variants were not associated with gallstone risk. Conclusions The p.D19H ABCG8 and, to a lesser extent, NPC1L1 rs217434 variants increase the risk of early-onset gallstone formation. These results point to the presence of a common lithogenic pathway in children and adults

RegPrecise: a database of curated genomic inferences of transcriptional regulatory interactions in prokaryotes

The RegPrecise database (http://regprecise.lbl.gov) was developed for capturing, visualization and analysis of predicted transcription factor regulons in prokaryotes that were reconstructed and manually curated by utilizing the comparative genomic approach. A significant number of high-quality inferences of transcriptional regulatory interactions have been already accumulated for diverse taxonomic groups of bacteria. The reconstructed regulons include transcription factors, their cognate DNA motifs and regulated genes/operons linked to the candidate transcription factor binding sites. The RegPrecise allows for browsing the regulon collections for: (i) conservation of DNA binding sites and regulated genes for a particular regulon across diverse taxonomic lineages; (ii) sets of regulons for a family of transcription factors; (iii) repertoire of regulons in a particular taxonomic group of species; (iv) regulons associated with a metabolic pathway or a biological process in various genomes. The initial release of the database includes ∼11 500 candidate binding sites for ∼400 orthologous groups of transcription factors from over 350 prokaryotic genomes. Majority of these data are represented by genome-wide regulon reconstructions in Shewanella and Streptococcus genera and a large-scale prediction of regulons for the LacI family of transcription factors. Another section in the database represents the results of accurate regulon propagation to the closely related genomes