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Abstract Prostate-specific antigen (PSA) levels are widely used to screen for prostate cancer, yet the test has poor sensitivity, specificity and predictive value, which leads to overdiagnosis and overtreatment. Alterations in the glycosylation status of PSA, including fucosylation, may offer scope for an improved biomarker. We sought to generate a monoclonal antibody (mAb) targeting α-1,6-fucosylated PSA (fuc-PSA) and to develop a tissue-based immunological assay for fuc-PSA detection. Immunogens representing fuc-PSA were used for immunisation and resultant mAbs were extensively characterised. The mAbs reacted specifically with fuc-PSA-specific glycopeptide, but not with aglycosylated PSA or glycan without the PSA peptide. Reactivity was confirmed using high-throughput surface plasmon resonance spectroscopy. X-ray crystallography investigations showed that the mAbs bound to an α-helical form of the peptide, whereas the native PSA epitope is linear. Protein unfolding was required for detection of fuc-PSA in patient samples. Peptide inhibition of fuc-PSA mAbs was observed with positive screening reagents, and target epitope specificity was observed in formalin-fixed, paraffin-embedded tissue samples. This research introduces a well-characterised, first-in-class antibody targeting fuc-PSA and presents the first crystal structure of an antibody demonstrating glycosylation-specific binding to a peptide.

This study explores a new eco-friendly approach for developing bioactive gelatin films using UV-C irradiation-induced photo-crosslinking. Riboflavin, a food-grade photoinitiator, was selected at an optimal concentration of 1.25% (w/w) for crosslinking gelatin under UV-C exposure for 4 to 22 min. Physicochemical analyses revealed enhanced tensile strength, reduced water vapor permeability, and lower water solubility in films crosslinked for up to 13 min. FTIR analysis demonstrated significant molecular changes, confirming the formation of crosslinking connections in gelatin–riboflavin films. Antimicrobial nanoemulsion (NE) (0.5, 0.75, 1% v/v) was incorporated into crosslinked films and applied to fresh beef. The 1% NE film exhibited the strongest antimicrobial effect, extending shelf-life by 20 days. In vitro release study confirmed Fickian diffusion behavior in the 1% NE film. This study also investigated the synergy between 1% NE film and three different types of modified atmosphere packaging (MAP) on the microbiological and physicochemical properties of beef for 26 days. The best results were achieved with 1% NE film under MAP1 and MAP2, which preserved meat redness and prevented lipid oxidation, extending the shelf-life up to 26 days. Therefore, UV-C irradiation-induced crosslinked bioactive film combined with high-oxygen MAP offers a promising solution for prolonging the shelf-life of beef.

Freezing rain is a rare and extremely dangerous meteorological phenomenon, consisting of the fall of liquid precipitation, while the air temperature at ground level falls below 0oC, favoring the freezing of raindrops and forming an ice sheet on the surfaces on which they are deposited. In Romania, the areas affected by the freezing rain phenomenon undergo significant changes regarding the structure and composition of the forest vegetation. The main purpose of this work is to figure out how to manage the forest stands affected by the freezing rain identifying optimal solutions for ecological reconstruction and prevention of significant damages. For the research of the affected areas, we placed 7 experimental areas, having the size of 500 m2 each, all the trees that were in this radius were inventoried. In this research work we evaluate the presence of this phenomenon which was reported in the Stejarul Zalău Forest District, in 2014, then in 2017, but with a lower intensity. Out of a total of 1627.6 ha, 500 ha were affected by the freezing rain, most of them being surfaces with beech forest stands with a consistency of 0.9, having a high slenderness index. Following the results obtained, it was observed that the most affected trees were the youngest. These having a reduced diameter, thin branches and a high slenderness coefficient.

Age-related cerebromicrovascular endothelial dysfunction underlies the initiation and progression of cognitive dysfunction and dementia, thus increasing the susceptibility of older adults to such conditions. Normal brain function requires dynamic adjustment of cerebral blood flow to meet the energetic demands of active neurons, which is achieved the homeostatic mechanism neurovascular coupling (NVC). In this context, therapeutical strategies aimed at rescuing or preserving NVC responses can delay the incidence or mitigate the severity of age-related cognitive dysfunction, and time-restricted eating (TRE) is a potential candidate for such a strategy. Studies have reported that TRE can improve cardiometabolic risk factors in older adults. However, the effect of TRE on cerebrovascular endothelial function remains unexplored. Thus, this protocol outlines the study procedures to test our hypothesis that a 6-month TRE regimen of 10-h eating window will improve NVC responses and endothelial function in community-dwelling older adults. This is a single-arm, open-label interventional trial. We aim to recruit 32 adults aged 55-80 years. Participants are instructed to maintain a TRE regimen of 10 h of free eating followed by 14 h of fasting for 6 months. Before and after fasting, participants are assessed for cognitive performance, peripheral micro- and macrovascular endothelial function, and NVC responses, as well as for several confounding factors, including body composition, dietary, and physical activity data. We expect that 6 months of TRE will improve NVC response and endothelial function in older adults compared with baseline, and that these improvements will be accompanied by improvements in cognitive performance. The study proposed herein will provide critical insight into a new potential therapeutical strategy for targeting age-related cognitive dysfunction. Ultimately, slowing down or alleviating cognitive decline will translate into improved quality of life and longer healthspan for aging adults. This study was prospectively registered at ClinicalTrials.gov (NCT06019195) on August 24, 2023.

Abstract De-oiled sunflower meal (DSF) and its protein isolate were evaluated as emulsifiers to replace egg yolk powder (EYP) in cookies. The chemical emulsifier DATEM (Diacetyl Tartaric Acid Esters of Mono- and Diglycerides) was used as a positive control. An experimental design of mixtures of the simplex-centroid type was carried out, and the ingredients were expressed as pseudo-components for EYP, DSF, and DATEM emulsifier. The DSF and its sunflower protein isolate (SPI) were tested to validate the design in optimized conditions. Whole meal cookies were analyzed in relation to rheological, physical, technological, and sensory characteristics using the control difference test. In the rheology of the dough, the DSF caused a reduction in the value of hardness, while increasing the parameter of elasticity. Instrumental texture results as well as the specific volume of the samples showed no difference. The control difference test regarding the cookies made with EYP, SPI, and DSF showed that consumers did not give different ratings to cookies made with sunflower as an emulsifier. Therefore, according to the parameters, conditions, and analysis performed, the replacement of EYP by DSF and SPI proved to be satisfactory as an emulsifying agent regarding the preparation of cookies for vegan consumers.

The dessert banana is a popular fruit worldwide, but its ripening process is greatly accelerated by high temperatures, which eventually leads to an unpleasant taste and the appearance of spots on the skin of the fruits. To slow down the ripening of bananas, expensive strategies are used, which are usually not practical for conventional farmers in less developed countries. In this study, we try to find a less costly alternative. Therefore, the effects of coatings of lemon peel extract (2.5%, 5%, and 10%), calcium chloride (4%), and glycerol (2%) on the shelf life and postharvest quality of the banana fruit (Cavendish) stored at 19–22 °C and 40–60% relative humidity were investigated. Treatment with a mixture of 2.5% lemon peel extract and 2% glycerol resulted in an extension of the shelf life of the dessert banana by up to 6 days and no detectable fungal infestation. The coating solution is an effective alternative to extend the shelf life and reduce quality losses in bananas.

In the era of immune checkpoint inhibitors, understanding the metastatic microenvironment of proficient mismatch repair/microsatellite stable (pMMR/MSS) colorectal cancer (CRC) is of paramount importance to both prognostication and the development of more effective novel therapies. In this study, primary and paired metastasis tissue samples were collected from patients with resectable metastatic CRC treated with adjuvant FOLFOX or peri‐operative chemotherapy in the MIROX phase III prospective study. In total, 74 cancer tissues were stained for CD3, CD8, Forkhead box protein 3 (FOXP3), programmed cell death protein‐1 (PD‐1, invasive front, stromal, intra‐epithelial compartments), and programmed death‐ligand 1 (PD‐L1, tumor, immune cells). The immune profiling of primary CRC had a limited value to predict the immune context of paired metastases for all markers but CD3+. The expression of CD8 and PD‐L1 was higher in metastases after neoadjuvant FOLFOX. In metastases, both CD3 T cells at the invasive front and PD‐L1 expressions on immune cells were predictive of better disease‐free survival. These results show that the effect of FOLFOX on modifying the immune microenvironment in resected CRC metastases and measurement of PD‐L1 expression and tumor‐infiltrating CD8 T cells in pMMR/MSS metastatic tissue samples could improve treatment strategies of metastatic CRC patients.

The Gram-negative bacterium Neisseria meningitidis, which causes meningitis in humans, has been demonstrated to manipulate or alter host signalling pathways during infection of the central nervous system (CNS). However, these complex signalling networks are not completely understood. We investigate the phosphoproteome of an in vitro model of the blood-cerebrospinal fluid barrier (BCSFB) based on human epithelial choroid plexus (CP) papilloma (HIBCPP) cells during infection with the N. meningitidis serogroup B strain MC58 in presence and absence of the bacterial capsule. Interestingly, our data demonstrates a stronger impact on the phosphoproteome of the cells by the capsule-deficient mutant of MC58. Using enrichment analyses, potential pathways, molecular processes, biological processes, cellular components and kinases were determined to be regulated as a consequence of N. meningitidis infection of the BCSFB. Our data highlight a variety of protein regulations that are altered during infection of CP epithelial cells with N. meningitidis, with the regulation of several pathways and molecular events only being detected after infection with the capsule-deficient mutant. Mass spectrometry proteomics data are available via ProteomeXchange with identifier PXD038560.

A tanulmányban egy nemzetközi kutatás (Myplace) adatait felhasználva, politikai szocializációs szempontból lényegesen eltérő két városban (Ózd és Sopron) élő 15–26 éves fiatalok radikalizmus értelmezését a szociális reprezentáció elmélet alapján vizsgáltuk. Az elméletre épülő egyik empirikus módszernek megfelelően, a 2012-ben lebonyolított kérdőíves adatfelvétel során asszociációs válaszok kvantifikálásával a radikalizmus reprezentációjának strukturális és tartalmi jellemzését követően a fiatalokra jellemző nacionalizmus és a demokratikus alapelvekhez való viszony, valamint a radikalizmus reprezentációjának kapcsolatát elemeztük.

Summary: Choroid plexus, located in brain ventricles, is the site of blood-cerebrospinal fluid barrier that contains endothelial cells and an epithelial monolayer separated by stroma. We established a two-cell-type model of the human choroid plexus consisting of immortalized endothelial cells (iHCPEnC) and epithelial papilloma (HIBCPP) cells grown on opposite sides of filter supports. In this protocol, we describe the preparation of this model, the measurement of transepithelial electrical resistance (TEER), and immunofluorescence imaging-based analysis to determine the barrier function.For complete details on the use and execution of this protocol, please refer to Muranyi et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Spinal muscular atrophy (SMA) is a progressive neuromuscular disease caused by biallelic pathogenic/likely pathogenic variants of the survival motor neuron 1 (SMN1) gene. Early diagnosis via newborn screening (NBS) and pre-symptomatic treatment are essential to optimize health outcomes for affected individuals. We developed a multiplex quantitative polymerase chain reaction (qPCR) assay using dried blood spot (DBS) samples for the detection of homozygous absence of exon 7 of the SMN1 gene. Newborns who screened positive were seen urgently for clinical evaluation. Confirmatory testing by multiplex ligation-dependent probe amplification (MLPA) revealed SMN1 and SMN2 gene copy numbers. Six newborns had abnormal screen results among 47,005 newborns screened during the first year and five were subsequently confirmed to have SMA. Four of the infants received SMN1 gene replacement therapy under 30 days of age. One infant received an SMN2 splicing modulator due to high maternally transferred AAV9 neutralizing antibodies (NAb), followed by gene therapy at 3 months of age when the NAb returned negative in the infant. Early data show that all five infants made excellent developmental progress. Based on one year of data, the incidence of SMA in Alberta was estimated to be 1 per 9401 live births.

INTRODUCTION: Mild cognitive impairment (MCI) is a prodromal stage of dementia. Understanding the mechanistic changes from healthy aging to MCI is critical for comprehending disease progression and enabling preventative intervention. METHODS: Patients with MCI and age‐matched controls (CN) were administered cognitive tasks during functional near‐infrared spectroscopy (fNIRS) recording, and changes in plasma levels of extracellular vesicles (EVs) were assessed using small‐particle flow cytometry. RESULTS: Neurovascular coupling (NVC) and functional connectivity (FC) were decreased in MCI compared to CN, prominently in the left‐dorsolateral prefrontal cortex (LDLPFC). We observed an increased ratio of cerebrovascular endothelial EVs (CEEVs) to total endothelial EVs in patients with MCI compared to CN, correlating with structural MRI small vessel ischemic damage in MCI. LDLPFC NVC, CEEV ratio, and LDLPFC FC had the highest feature importance in the random Forest group classification. DISCUSSION: NVC, CEEVs, and FC predict MCI diagnosis, indicating their potential as markers for MCI cerebrovascular pathology. Highlights: Neurovascular coupling (NVC) is impaired in mild cognitive impairment (MCI).Functional connectivity (FC) compensation mechanism is lost in MCI.Cerebrovascular endothelial extracellular vesicles (CEEVs) are increased in MCI.CEEV load strongly associates with cerebral small vessel ischemic lesions in MCI.NVC, CEEVs, and FC predict MCI diagnosis over demographic and comorbidity factors. [ABSTRACT FROM AUTHOR]

Pea protein isolate (PPI, from Pisum sativum L.) was fermented with six different lactic acid bacteria strains for 24 ​h and 48 ​h. The fermented samples were analyzed regarding their retronasal aroma and taste, their protein solubility, emulsifying and foaming capacity. Changes in the molecular weight distribution were analyzed to monitor potential effects of fermentation on the main allergenic protein fractions of PPI. After 24-h fermentation, PPI's characteristic aroma attributes and bitter taste decreased for all fermented PPI. However, after 48-h fermentation, cheesy aroma, and acid and salty tastes were increased. The PPI fermented with L. plantarum showed the most neutral taste and the panel's highest preference; instead, fermentation with L. fermentum led to a fecal aroma and was the least preferred. The protein solubility and emulsifying capacity decreased after PPI fermentation, while foaming capacity remained constant in comparison to the untreated PPI. The electrophoretic results showed a reduction in the intensity of the allergenic protein fractions; however, these changes might be attributed to the reduced protein solubility rather than to a high proteolytic effect of the strains. Fermentation of PPI for 24 ​h and 48 ​h might not be a suitable method for the production of highly functional pea proteins. Further modification methods have to be investigated in the future.

Summary: The choroid plexus (CP) is a highly vascularized structure containing endothelial and epithelial cells located in the ventricular system of the central nervous system (CNS). The role of the fenestrated CP endothelium is under-researched and requires the generation of an immortalized CP endothelial cell line with preserved features. Transduction of primary human CP endothelial cells (HCPEnC) with the human telomerase reverse transcriptase (hTERT) resulted in immortalized HCPEnC (iHCPEnC), which grew as monolayer with contact inhibition, formed capillary-like tubes in Matrigel, and showed no colony growth in soft agar. iHCPEnC expressed pan-endothelial markers and presented characteristic plasmalemma vesicle-associated protein-containing structures. Cultivation of iHCPEnC and human epithelial CP papilloma (HIBCPP) cells on opposite sides of cell culture filter inserts generated an in vitro model with a consistently enhanced barrier function specifically by iHCPEnC. Overall, iHCPEnC present a tool that will contribute to the understanding of CP organ functions, especially endothelial-epithelial interplay.

Combinations of enzymatic hydrolysis using different proteolytic enzymes (papain, Esperase®, trypsin) and lactic fermentation with Lactobacillus plantarum were used to alter potential pea allergens, the functional properties and sensory profile of pea protein isolate (PPI). The order in which the treatments were performed had a major impact on the changes in the properties of the pea protein isolate; the highest changes were seen with the combination of fermentation followed by enzymatic hydrolysis. SDS-PAGE, gel filtration, and ELISA results showed changes in the protein molecular weight and a reduced immunogenicity of treated samples. Treated samples showed significantly increased protein solubility at pH 4.5 (31.19–66.55%) and at pH 7.0 (47.37–74.95%), compared to the untreated PPI (6.98% and 40.26%, respectively). The foaming capacity was significantly increased (1190–2575%) compared to the untreated PPI (840%). The treated PPI showed reduced pea characteristic off-flavors, where only the treatment with Esperase® significantly increased the bitterness. The results from this study suggest that the combination of enzymatic hydrolysis and lactic fermentation is a promising method to be used in the food industry to produce pea protein ingredients with higher functionality and a highly neutral taste. A reduced detection signal of polyclonal rabbit anti-pea-antibodies against the processed protein preparations in ELISA furthermore might indicate a decreased immunological reaction after consumption.

The application of essential oils as antimicrobials is a current subject of research and a promising approach in terms of natural food preservation. Due to the diversity of EO producing plant genera and the inconsistent use of susceptibility testing methods, information on the antibacterial potency of many EO varieties is fragmentary. This study was performed to assess the minimal inhibitory concentrations (MIC) of 179 EO samples from 86 plant varieties, using a single method approach, excluding emulsifying agents. MICs were acquired in a broth microdilution assay, using a dispersion based approach to incorporate EOs in a concentration range of 6400 to 50 μg/ml. Staphylococcus aureus and Escherichia coli were used as model bacteria. At concentrations below 400 μg/ml S. aureus was inhibited by 30, E. coli by 12 EO varieties. Azadirachta indica (50 μg/ml vs. S. aureus) and Litsea cubeba (50 μg/ml vs. S. aureus, 200 μg/ml vs. E. coli) essential oils were identified as promising new antimicrobial EO candidates with significant antimicrobial activity against the two foodborne pathogenic bacteria.

The generation of the high species diversity of insects in Japan was profoundly influenced by the formation of the Japanese Archipelago. We explored the species diversification and biogeographical history of the Nemouridae Billberg, 1820 family in the Japanese Archipelago using mitochondrial DNA and nuclear DNA markers. We collected 49 species among four genera: Indonemoura Baumann, 1975; Protonemura Kempny, 1898; Amphinemura, Ris 1902 and Nemoura Latreille, 1796 in Japan, China, South Korea and North America. We estimated their divergence times-based on three molecular clock node calibrations-using Bayesian phylogeography approaches. Our results suggested that Japanese Archipelago formation events resulted in diversification events in the middle of the Cretaceous (

Pea protein concentrates and isolates are important raw materials for the production of plant-based food products. To select suitable peas (Pisum sativum L.) for protein extraction for further use as food ingredients, twelve different cultivars were subjected to isoelectric precipitation and spray drying. Both the dehulled pea flours and protein isolates were characterized regarding their chemical composition and the isolates were analyzed for their functional properties, sensory profiles, and molecular weight distributions. Orchestra, Florida, Dolores, and RLPY cultivars showed the highest protein yields. The electrophoretic profiles were similar, indicating the presence of all main pea allergens in all isolates. The colors of the isolates were significantly different regarding lightness (L*) and red-green (a*) components. The largest particle size was shown by the isolate from Florida cultivar, whereas the lowest was from the RLPY isolate. At pH 7, protein solubility ranged from 40% to 62% and the emulsifying capacity ranged from 600 to 835 mL g−1. The principal component analysis revealed similarities among certain pea cultivars regarding their physicochemical and functional properties. The sensory profile of the individual isolates was rather similar, with an exception of the pea-like and bitter attributes, which were significantly different among the isolates.

Dieser Artikel beschäftigt sich mit dem in Ungarn geborenen Philologen, Didaktiker, Lexikologen und Übersetzer Paulo Rónai, der 1941 nach Brasilien emigrierte. Rónais Werdegang und seine fast lebenslange Beziehung zu Brasilien werden als Übersetzungsgesamtleistung betrachtet, bei der professionelle und biographische Aspekte nicht getrennt voneinander gedacht werden können: Übersetzen und Über-setzen in einen anderen Sprach- und Kulturraum verschränken und bedingen sich.

Verzehrfertige Salate, Obstprodukte und Sprossen boomen. Allerdings bergen sie ein nicht unerhebliches Risiko einer Belastung mit Krankheitserregern. Warum lassen sich Einträge von pathogenen Bakterien trotz hoher Hygienestandards in den verarbeitenden Betrieben nicht vollständig vermeiden? An welchen Stellen der Warenketten erfolgen Kontaminationen und wie können sich die Erreger vermehren? Das vom BMBF im Rahmen des Programms „Forschung für die zivile Sicherheit“ geförderte Verbundprojekt SAFEFRESH hat erforscht, wie sich das mikrobielle Risiko bei Fresh-Cut-Erzeugnissen reduzieren lässt. Dazu wurden ein Monitoring- und verschiedene Entkeimungsverfahren entwickelt sowie realistische Szenarios zu möglichen Kontaminationsfällen und deren Folgekaskaden hergeleitet. Der Gewinn an zusätzlicher Sicherheit ist jedoch begrenzt.

In the present article we intend to point out the relationship between prejudice and national concepts in different national contexts among students. We tried to justify that the explanation based on the view of prejudices by Fuchs and Case - according to whom prejudice is not an attitude but a way of life – is valid in different national contexts. For this reason the procedure was applied in our survey in order to validate the conception in three (Hungarian, Finnish, Russian) different cultural-political contexts, and also to use it as a reference model in determining the explanation of prejudice. The second aim was to reveal the structural relationships between the national attitudes of the model and attitudes towards minority groups.

Sichere Lebensmittel von hoher Qualität stellen besonders bei leicht verderblichen Frischeprodukten eine Herausforderung für die Gestaltung der Nacherntekette dar. Da die Beprobung von Lebensmittelchargen in der Praxis meist anhand ausgewählter Indikatororganismen erfolgt, bleiben unerwartete, potenziell gefährliche Mikroorganismen häufig unentdeckt. Die Detektion dieser Bakterien ist jedoch von Interesse, um potenzielle Gefahren für den Verbraucher zu vermeiden. Am Beispiel von Mungobohnensprossen wurde die mikrobielle Diversität mittels Plattenzählverfahren und MALDI-TOF MS (matrix-assisted laser desorption/ionisation – time of flight mass spectrometry) ermittelt. Bei einer Gesamtkeimzahl zwischen 8 und 9 log KbE/g Sprossen konnten unter anderem Bakterien der Bacillus cereus Gruppe, Yersinia sp., Enterobacter spp., Klebsiella spp., Pantoea spp. und Pseudomonas spp. identifiziert werden.

The steps from HIV-1 cytoplasmic entry until integration of the reverse transcribed genome are currently enigmatic. They occur in ill-defined reverse-transcription- and pre-integration-complexes (RTC, PIC) with various host and viral proteins implicated. In this study, we report quantitative detection of functional RTC/PIC by labeling nascent DNA combined with detection of viral integrase. We show that the viral CA (capsid) protein remains associated with cytoplasmic RTC/PIC but is lost on nuclear PIC in a HeLa-derived cell line. In contrast, nuclear PIC were almost always CA-positive in primary human macrophages, indicating nuclear import of capsids or capsid-like structures. We further show that the CA-targeted inhibitor PF74 exhibits a bimodal mechanism, blocking RTC/PIC association with the host factor CPSF6 and nuclear entry at low, and abrogating reverse transcription at high concentrations. The newly developed system is ideally suited for studying retroviral post-entry events and the roles of host factors including DNA sensors and signaling molecules.

The inner structural Gag proteins and the envelope (Env) glycoproteins of human immunodeficiency virus (HIV-1) traffic independently to the plasma membrane, where they assemble the nascent virion. HIV-1 carries a relatively low number of glycoproteins in its membrane, and the mechanism of Env recruitment and virus incorporation is incompletely understood. We employed dual-color super-resolution microscopy visualizing Gag assembly sites and HIV-1 Env proteins in virus-producing and in Env expressing cells. Distinctive HIV-1 Gag assembly sites were readily detected and were associated with Env clusters that always extended beyond the actual Gag assembly site and often showed enrichment at the periphery and surrounding the assembly site. Formation of these Env clusters depended on the presence of other HIV-1 proteins and on the long cytoplasmic tail (CT) of Env. CT deletion, a matrix mutation affecting Env incorporation or Env expression in the absence of other HIV-1 proteins led to much smaller Env clusters, which were not enriched at viral assembly sites. These results show that Env is recruited to HIV-1 assembly sites in a CT-dependent manner, while Env(ΔCT) appears to be randomly incorporated. The observed Env accumulation surrounding Gag assemblies, with a lower density on the actual bud, could facilitate viral spread in vivo. Keeping Env molecules on the nascent virus low may be important for escape from the humoral immune response, while cell-cell contacts mediated by surrounding Env molecules could promote HIV-1 transmission through the virological synapse.

CpG-ODN stimulates dendritic cells (DCs) to produce cytokines, which are important for pathogenesis of autoimmune disorders and vaccine strategy for cancer. CpG-ODN activates the TLR9/MyD88/TRAF6 cascade leading to activation of IKK-NF-κB and JNK, which are critical for production of pro-inflammatory cytokines. However, whether other molecules are involved in activation of CpG-ODN signaling is still not clear. Here we report that the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) is involved in this activation process. DNA-PKcs-deficient DCs exhibited a defect in the IL-6 and IL-12 response to CpG-ODN in a dose- and time-dependent manner. Loss of DNA-PKcs impaired phosphorylation of IKK, IκBα, NF-κB and JNK in response to CpG-ODN. Interestingly, CpG-ODN was able to bind DNA-PKcs and induce its association and co-localization with TRAF6 in the absence of TLR9. Our data suggest that DNA-PKcs is a player in CpG-ODN signaling and may explain why DNA-PKcs is implicated in the pathogenic process of autoimmune disease.

Fluorescently labeled human immunodeficiency virus (HIV) derivatives, combined with the use of advanced fluorescence microscopy techniques, allow the direct visualization of dynamic events and individual steps in the viral life cycle. HIV proteins tagged with fluorescent proteins (FPs) have been successfully used for live-cell imaging analyses of HIV-cell interactions. However, FPs display limitations with respect to their physicochemical properties, and their maturation kinetics. Furthermore, several independent FP-tagged constructs have to be cloned and characterized in order to obtain spectral variations suitable for multi-color imaging setups. In contrast, the so-called SNAP-tag represents a genetically encoded non-fluorescent tag which mediates specific covalent coupling to fluorescent substrate molecules in a self-labeling reaction. Fusion of the SNAP-tag to the protein of interest allows specific labeling of the fusion protein with a variety of synthetic dyes, thereby offering enhanced flexibility for fluorescence imaging approaches.Here we describe the construction and characterization of the HIV derivative HIV(SNAP), which carries the SNAP-tag as an additional domain within the viral structural polyprotein Gag. Introduction of the tag close to the C-terminus of the matrix domain of Gag did not interfere with particle assembly, release or proteolytic virus maturation. The modified virions were infectious and could be propagated in tissue culture, albeit with reduced replication capacity. Insertion of the SNAP domain within Gag allowed specific staining of the viral polyprotein in the context of virus producing cells using a SNAP reactive dye as well as the visualization of individual virions and viral budding sites by stochastic optical reconstruction microscopy. Thus, HIV(SNAP) represents a versatile tool which expands the possibilities for the analysis of HIV-cell interactions using live cell imaging and sub-diffraction fluorescence microscopy.

The extremely radical (mostly youth) political organizations may be regarded as peculiar youth subcultures. We believe it is true not only for Hungary where relatively rich social scientific knowledge exists on the majority perception of such organizations. However, not many instances are found for the empirical sociological study of the organizations. The sample of the survey conducted in the autumn of 2010 by the Centre of Conflict Research at ELTE and the Sociology and Social Policy Department at the University of Debrecen consists of members of a right-wing paramilitary organization (The National Army of Guardsmen Heritage Protection and Civil Guard Organization) founded three years ago and, as control sample, the members of an environmental protection (non-radical, non-militant) organization (E-mission) operating in the same region (N=100-100 persons). The research focuses on new authoritarianism, which only partly rests on the “classical” authoritarianism, i.e. the socially determined pattern of the modern individual interiorized throughout socialization, as described by freudo-marxism in the beginning of the 20 th century. In developing the theoretical background and the operationalization, besides traditional dimensions and socializational characteristics, we relied on theories (e. g. social dominance) and topics (e. g. conflict perception and reduction) which have not been applied in such studies before. Based on interpretational-statistical models, this paper analyses the socio-cultural factors that shape the social dominance orientation characteristics for the two sub-samples.

In the era of immune checkpoint inhibitors, understanding the metastatic microenvironment of proficient mismatch repair/microsatellite stable (pMMR/MSS) colorectal cancer (CRC) is of paramount importance to both prognostication and the development of more effective novel therapies. In this study, primary and paired metastasis tissue samples were collected from patients with resectable metastatic CRC treated with adjuvant FOLFOX or peri-operative chemotherapy in the MIROX phase III prospective study. In total, 74 cancer tissues were stained for CD3, CD8, Forkhead box protein 3 (FOXP3), programmed cell death protein-1 (PD-1, invasive front, stromal, intra-epithelial compartments), and programmed death-ligand 1 (PD-L1, tumor, immune cells). The immune profiling of primary CRC had a limited value to predict the immune context of paired metastases for all markers but CD3+. The expression of CD8 and PD-L1 was higher in metastases after neoadjuvant FOLFOX. In metastases, both CD3 T cells at the invasive front and PD-L1 expressions on immune cells were predictive of better disease-free survival. These results show that the effect of FOLFOX on modifying the immune microenvironment in resected CRC metastases and measurement of PD-L1 expression and tumorinfiltrating CD8 T cells in pMMR/MSS metastatic tissue samples could improve treatment strategies of metastatic CRC patients. [ABSTRACT FROM AUTHOR]