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

Audio subtitling : voicing strategies and their effect on film enjoyment

Media Accessibility, understood as a branch of Audiovisual Translation studies devoted to the study of access services to media, has experienced a growth in research in recent years. Even if some of the techniques in this field have produced a great number of works, others are still underresearched. This article reports on a pilot study carried out within the NEA Project1. The aim of the study is to compare different AST delivery styles in an experiment during which data will be gathered through subjective and objective measures. This article reports on the results of a pilot study in which the research procedure was set up and tested. Our main findings show that both subjective and objective assessment methods are valid to evaluate emotional experience induced by film

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

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

Electrodermal activity as a measure of emotions in media accessibility research : methodological considerations

This article proposes electrodermal activity (EDA) as a new objective measure for experimental studies in media accessibility. It first presents a theoretical framework in which the concept of emotion and its categorisation are presented. It then explains how EDA can be used to measure emotional reaction: the article reports on experimental design, participant selection, stimuli preparation, data collection devices, experimental procedure and data analysis. It also discusses briefly how EDA can be combined with other measures. Overall, the article provides a general methodological framework for the implementation of electrodermal activity as a measure of emotions in media accessibility researc

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

Phototropin2 3’UTR overlaps with the AT5G58150 gene encoding an inactive RLK kinase

ackground This study examines the biological implications of an overlap between two sequences in the Arabidop�sis genome, the 3’UTR of the PHOT2 gene and a putative AT5G58150 gene, encoded on the complementary strand. AT5G58150 is a probably inactive protein kinase that belongs to the transmembrane, leucine-rich repeat receptor-like kinase family. Phot2 is a membrane-bound UV/blue light photoreceptor kinase. Thus, both proteins share their cellular localization, on top of the proximity of their loci. Results The extent of the overlap between 3’UTR regions of AT5G58150 and PHOT2 was found to be 66 bp, using RACE PCR. Both the at5g58150 T-DNA SALK_093781C (with insertion in the promoter region) and 35S::AT5G58150�GFP lines overexpress the AT5G58150 gene. A detailed analysis did not reveal any substantial impact of PHOT2 or AT5G58150 on their mutual expression levels in diferent light and osmotic stress conditions. AT5G58150 is a plasma membrane protein, with no apparent kinase activity, as tested on several potential substrates. It appears not to form homodimers and it does not interact with PHOT2. Lines that overexpress AT5G58150 exhibit a greater reduction in lat‑ eral root density due to salt and osmotic stress than wild-type plants, which suggests that AT5G58150 may partici‑ pate in root elongation and formation of lateral roots. In line with this, mass spectrometry analysis identifed proteins with ATPase activity, which are involved in proton transport and cell elongation, as putative interactors of AT5G58150. Membrane kinases, including other members of the LRR RLK family and BSK kinases (positive regulators of brassinos‑ teroid signalling), can also act as partners for AT5G58150. Conclusions AT5G58150 is a membrane protein that does not exhibit measurable kinase activity, but is involved in signalling through interactions with other proteins. Based on the interactome and root architecture analysis, AT5G58150 may be involved in plant response to salt and osmotic stress and the formation of roots in Arabidopsis

TP53 polymorphism in plasma cell myeloma

Introduction. Significant and accessible predictive factors for bortezomib treatment in plasma cell myeloma (PCM) are still lacking. TP53 codon 72 polymorphism (P72R) results in proline (P) or arginine (R) at 72 amino acid position, which causes synthesis of proteins with distinct functions. The aims of our study were to: 1) analyze whether this polymorphism is associated with an increased risk of PCM; 2) study whether the P72R polymorphism affects overall survival (OS) among PCM patients; 3) assess the possible association of the P72R polymorphism with sensitivity to bortezomib in cell cultures derived from PCM patients. Material and methods. Genomic DNA from newly diagnosed 59 patients (without IgVH gene rearrangements and TP53 deletions) and 50 healthy blood donors were analyzed by RFLP-PCR to identify TP53 polymorphism. Chromosomal aberrations were detected by use of cIg-FISH. The lymphocyte cell cultures from a subgroup of 40 PCM patients were treated with bortezomib (1, 2 and 4 nM). Results. The P allele of the P72R polymorphism was more common than the R allele in PMC patients compared to controls (39% vs. 24%), and the difference was significant (p = 0.02). The PP and PR genotypes (in combina­tion) were more frequent among cases than in controls (65% vs. 42%, OR = 2.32, p = 0.04). At the cell culture level and 2 nM bortezomib concentration the PP genotype was associated with higher necrosis rates (10.5%) compared to the PR genotype (5.7%, p = 0.006) or the RR genotype (6.3%, p = 0.02); however, no effect of genotypes was observed at bortezomib concentrations of 1 and 4 nM. The shortest OS (12 months) was observed in patients with the PP genotype compared to patients with the PR or RR genotypes (20 months) (p = 0.04). Conclusions. The results suggest that P72R polymorphisms may be associated with an increased PCM risk and may affect OS of PCM patients. However, we saw no consistent results of the polymorphism effect on apoptosis and necrosis in cell cultures derived from PCM patients. Further studies are need in this regard