Mikroszatellita-instabilitás előfordulása, intratumoralis heterogenitása, prognosztikus és prediktív potenciálja primer colorectalis carcinomák és párosított májáttéteik sebészi kezelését követően

Absztrakt Bevezetés: A vastagbéldaganatokra jellemző genetikai instabilitás megnyilvánulhat több úton: kromoszomális instabilitás, mikroszatellita-instabilitás, illetve „CpG-island methylator phenotype”. Ezek pontosabb karakterizálásával a rendelkezésre álló kezelések elviekben optimalizálhatók lehetnek. Célkitűzés: A szerzők a mikroszatellita-instabilitás előfordulását, heterogenitását, prognosztikus és prediktív potenciálját vizsgálták 122 primer colontumor szisztematikusan szelektált régióiban és 69 párosított májmetasztázisban. Módszer: Szöveti multiblokkok kialakítása után az MLH1, MSH2, MSH6 és PMS2 kifejeződését vizsgálták immunhisztokémiai módszerrel. Eredmények: A betegek 11,5%-a (14/122) rendelkezett mikroszatellita-instabil fenotípusú daganattal. A különböző tumorrégiók fehérjekifejeződésében nem volt jelentős különbség. A primer tumor–májmetasztázis párok esetében 20,2%-ban a kettő más mismatch repair státusba volt sorolható. A relapsusmentes és teljes túlélést tekintve a mismatch repair státus nem volt prognosztikus. Az 5-fluorouracil-, oxaliplatin-, irinotecan-, bevacizumab-, cetuximab-, panitumumabterápia hatékonyságát tekintve mismatch repair státus nem volt prediktív a progressziómentes és teljes túlélés adatai alapján. Következtetések: A prognosztikus faktorok pontosabb meghatározása nagyobb esetszámú, pontosan szelektált vizsgálat keretében hatékonyabbá teheti a kezelés megválasztását. Orv. Hetil., 2015, 156(36), 1460–1471

The psychological consequences of the ecological crisis: Three new questionnaires to assess eco-anxiety, eco-guilt, and ecological grief

As climate change develops reactions such as eco-anxiety, eco-guilt and ecological grief are becoming increasingly common. Our aim was to develop questionnaires to assess these psychological consequences, and to examine their relationship with pro-environmental behavior (PEB). Items of the questionnaires were generated based on literature review and the qualitative analysis of semi-structured interviews (N = 17). The first item pool was administered to a large adult sample (N = 4608) along with assessing PEB. The Eco-Guilt Questionnaire (EGuiQ-11) and the Ecological Grief Questionnaire (EGriQ-6) each had a one-factor structure, while the Eco-Anxiety Questionnaire (EAQ-22) consisted of two factors: habitual ecological worry and the negative consequences of eco-anxiety. The factors were positively associated with PEB. The questionnaires had a robust factor structure, and they are suitable for the assessment of a wide range of negative emotional states related to climate change and the ecological crisis. Our results indicate the possible utility of negative emotions in reinforcing PEB

Herpesvirus-mediated delivery of a genetically encoded fluorescent Ca2+ sensor to canine cardiomyocytes

We report the development and application of a pseudorabies virus-based system for delivery of troponeon, a fluorescent Ca2+ sensor to adult canine cardiomyocytes. The efficacy of transduction was assessed by calculating the ratio of fluorescently labelled and nonlabelled cells in cell culture. Interaction of the virus vector with electrophysiological properties of cardiomyocytes was evaluated by the analysis of transient outward current (Ito), kinetics of the intracellular Ca2+ transients, and cell shortening. Functionality of transferred troponeon was verified by FRET analysis. We demonstrated that the transfer efficiency of troponeon to cultured adult cardiac myocytes was virtually 100%. We showed that even after four days neither the amplitude nor the kinetics of the Ito current was significantly changed and no major shifts occurred in parameters of [Ca2+]i transients. Furthermore, we demonstrated that infection of cardiomyocytes with the virus did not affect the morphology, viability, and physiological attributes of cells

Diclofenac Prolongs Repolarization in Ventricular Muscle with Impaired Repolarization Reserve

Background: The aim of the present work was to characterize the electrophysiological effects of the non-steroidal anti- inflammatory drug diclofenac and to study the possible proarrhythmic potency of the drug in ventricular muscle. Methods: Ion currents were recorded using voltage clamp technique in canine single ventricular cells and action potentials were obtained from canine ventricular preparations using microelectrodes. The proarrhythmic potency of the drug was investigated in an anaesthetized rabbit proarrhythmia model. Results: Action potentials were slightly lengthened in ventricular muscle but were shortened in Purkinje fibers by diclofenac (20 mM). The maximum upstroke velocity was decreased in both preparations. Larger repolarization prolongation was observed when repolarization reserve was impaired by previous BaCl 2 application. Diclofenac (3 mg/kg) did not prolong while dofetilide (25 mg/kg) significantly lengthened the QT c interval in anaesthetized rabbits. The addition of diclofenac following reduction of repolarization reserve by dofetilide further prolonged QT c . Diclofenac alone did not induce Torsades de Pointes ventricular tachycardia (TdP) while TdP incidence following dofetilide was 20%. However, the combination of diclofenac and dofetilide significantly increased TdP incidence (62%). In single ventricular cells diclofenac (30 mM) decreased the amplitude of rapid (I Kr ) and slow (I Ks ) delayed rectifier currents thereby attenuating repolarization reserve. L-type calcium current (I Ca ) was slightly diminished, but the transient outward (I to ) and inward rectifier (I K1 ) potassium currents were not influenced. Conclusions: Diclofenac at therapeutic concentrations and even at high dose does not prolong repolarization markedly and does not increase the risk of arrhythmia in normal heart. However, high dose diclofenac treatment may lengthen repolarization and enhance proarrhythmic risk in hearts with reduced repolarization reserve

In depth evaluation of the prognostic and predictive utility of PTEN immunohistochemistry in colorectal carcinomas: performance of three antibodies with emphasis on intracellular and intratumoral heterogeneity.

BACKGROUND: Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) loss of function is frequently detected in advanced colorectal cancer. Its detection is thought to have prognostic significance and it is being considered to predict responsiveness to anti-EGFR therapy. Unfortunately, while immunohistochemical assessment of PTEN expression is widespread, it lacks standardization and the results are hardly comparable across the available publications. METHODS: Retrospectively collected, formalin-fixed and paraffin-embedded colorectal tumor tissue samples from 55 patients were combined into tissue microarray (TMA) blocks. We used three different PTEN antibodies to determine the frequency, intensity and intracellular pattern of PTEN immunohistochemical labeling: Neomarkers, Dako and CellSignaling. We evaluated the aforementioned parameters in selected regions of colorectal cancers and in their lymph node metastases by using three scoring methods that take into consideration both staining frequency and intensity (H1-H3-score). We also evaluated intracellular localization. RESULTS: The Dako and CellSignaling antibodies stained predominantly cytoplasms, while the Neomarkers antibody specifically stained cell nuclei. PTEN H-scores were significantly lower in all tumor areas as compared to the normal colonic mucosa based on staining with the DAKO and CellSignaling antibodies. Intratumoral regional differences or differences between matching tumors and metastases were not detected with any of the antibodies. Neither Dako, neither CellSignaling, nor the Neomarkers antibodies revealed a significant correlation between PTEN expression and pT, Dukes/MAC and clinical stage. KRAS status, histological grade correlated with PTEN H-scores based on staining with the Neomarkers antibody. PTEN H-scores did not correlate with MMR status. PTEN H-scores did not show any correlation with relapse-free survival based on staining with either antibody. CONCLUSIONS: While PTEN expression decreased in colorectal cancer according to two antibodies, neither of the three applied PTEN antibodies could justify significant correlation with clinicopathological data, nor had prognostic value. Thus, we might conclude that immunohistochemical PTEN investigation remains a challenge requiring more standardized evaluation on larger number of cases to clarify its utility as a prognostic and predictive tool in CRC. The standardization of immunohistochemical method is key in the evaluation process, which is further discussed

Predictors of conservation value of Turkish cemeteries: A case study using orchids

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In vitro dissolution–permeation evaluation of an electrospun cyclodextrin-based formulation of aripiprazole using μFlux™

Since it is a well-known fact that among the newly discovered active pharmaceutical ingredients the number of poorly water soluble candidates is continually increasing, dissolution enhancement of poorly water soluble drugs has become one of the central challenges of pharmaceutical studies. So far the preclinical studies have been mainly focused on formulation methods to enhance the dissolution of active compounds, in many cases disregarding the fact that the formulation matrix not only affects dissolution but also has an effect on the transport through biological membranes, changing permeation of the drug molecules. The aim of this study was to test an electrospun cyclodextrin-based formulation of aripiprazole with the novel μFlux apparatus, which monitors permeation together with dissolution, and by this means better in vitro–in vivo correlation is achieved. It was evinced that a cyclodextrin-based electrospun formulation of aripiprazole has the potential to ensure fast drug delivery through the oral mucosa owing to the ultrafast dissolution of the drug from the formulation and the enhanced flux across membranes as shown by the result of the novel in vitro dissolution and permeation test

Investigating the Prognostic Relevance of Tumor Immune Microenvironment and Immune Gene Assembly in Breast Carcinoma Subtypes

We hypothesized that different BC subtypes are characterized by spatially distinct tumor immune microenvironment (TIME) and that immune gene assembly of metastatic (Met) and non-metastatic (Ctrl) BCs vary across subtypes. Peritumoral, stromal and intratumoral TIL was assessed on 309 BC cases. Hot, cold and immune-excluded groups were defined, and the prognostic role of this classification was assessed. CD4+/CD8+ positivity was analyzed in 75 cases in four systematically predefined tumor regions. Immune gene expression of Met and Ctrl HER2-negative BCs was compared by using NanoString nCounter technology. The amount of TIL infiltration varied greatly within all BC subtypes. Two-third of the cases were cold tumors with no significant survival difference compared to hot tumors. A lower CD4+/CD8+ ratio at the stromal internal tumor region was significantly associated with longer distant metastasis-free survival. The differentially expressed immune genes between Met and Ctrl varied across the studied BC subtypes with TNBC showing distinct features from the luminal subtypes. The TIME is characterized by a considerable heterogeneity; however, low level of TILs does not equate to disease progression. The differences in immune gene expression observed between Met and Ctrl breast carcinomas call attention to the important role of altered immune function in BC progression

New thousand-seed weight dataset for plant species of Central Europe

One of the most important and most easily measurable physical characteristics of plant seeds is their weight, which influences and indicates crucial ecological processes. Seed weight affects spatial and temporal dispersibility, and can also influence seed predation and the germination, growth and survival of seedlings. Providing trait data for species missing from international databases is key to promote studies that advance our understanding of the functioning of plant communities and ecosystems, which is an essential issue in the face of the global climate change and biodiversity loss. Compared to species from Western and Northwestern Europe, those with an Eastern or Central European centre of distribution are underrepresented in most international trait databases. Therefore, the creation of specific trait databases is key to help regional studies. In this respect, it is important not only to collect fresh seeds for weight measurements, but also to measure and process data of seeds preserved in collections and make them available to the broader scientific community. In this data paper we provide seed weight data to fill in missing trait data of plant species of Central and Eastern Europe. Our dataset includes weight measurement for 281 taxa of the Central European flora including also some cultivated and exotic species. The seeds were collected between 1971 and 2021 mostly in Central Europe. One part of the measured seeds was collected in the last decade, the other part is from an older seed collection, but all seeds were measured recently. For each species, we collected a minimum of 3 × 100 intact seeds, if possible. The seeds were air-dried at room temperature (approximately 21 °C and 50% relative humidity) for at least two weeks and measured with an accuracy of 0.001 g using an analytical balance. The thousand-seed weights reported here were calculated based on the measured values. Our goal for the future is to incorporate the seed weight data reported here in a regional database (Pannonian Database of Plant Traits – PADAPT) that gathers plant traits and other plant characteristics for the Pannonian flora. The data presented here will facilitate trait-based analyses of the flora and vegetation of Central Europe