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As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm (Perinereis aibuhitensis) was expressed in E. coli (Escherichia coli) to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed in E. coli with low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein's solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm in E. coli and broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease. [ABSTRACT FROM AUTHOR]

Mitochondria provide cells with energy and regulate the cellular metabolism. Almost all mitochondrial proteins are nuclear-encoded, translated on ribosomes in the cytoplasm, and subsequently transferred to the different subcellular compartments of mitochondria. Here, we developed OptoMitoImport, an optogenetic tool to control the import of proteins into the mitochondrial matrix via the presequence pathway on demand. OptoMitoImport is based on a two-step process: first, light-induced cleavage by a TEV protease cuts off a plasma membrane-anchored fusion construct in close proximity to a mitochondrial targeting sequence; second, the mitochondrial targeting sequence preceding the protein of interest recruits to the outer mitochondrial membrane and imports the protein fused to it into mitochondria. Upon reaching the mitochondrial matrix, the matrix processing peptidase cuts off the mitochondrial targeting sequence and releases the protein of interest. OptoMitoImport is available as a two-plasmid system as well as a P2A peptide or IRES sequence-based bicistronic system. Fluorescence studies demonstrate the release of the plasma membrane-anchored protein of interest through light-induced TEV protease cleavage and its localization to mitochondria. Cell fractionation experiments confirm the presence of the peptidase-cleaved protein of interest in the mitochondrial fraction. The processed product is protected from proteinase K treatment. Depletion of the membrane potential across the inner mitochondria membrane prevents the mitochondrial protein import, indicating an import of the protein of interest by the presequence pathway. These data demonstrate the functionality of OptoMitoImport as a generic system with which to control the post-translational mitochondrial import of proteins via the presequence pathway. [ABSTRACT FROM AUTHOR]

• The article introduces a novel semi-empirical model for a TEV operating with R290. • No prior experimental work was found involving TEV operating with R290 in DX-SAHP. • The article emphasizes a careful data collection process encompassing 150 experiments. • The multiple linear regression resulted in a R² of 0.9648 with predictions not exceeding more than 15 % deviation. • The model was applied and compared with 44 experiments from the literature, resulting in a deviation of less than 15 %. The expansion device plays a crucial role in the operation of heat pumps and refrigeration systems, making its study integral to advancements in the fields of refrigeration and heating. Among various expansion devices, the Thermostatic Expansion Valve (TEV) stands out due to its rapid response time and mechanical nature, which grants it high control capacity over refrigerant superheating. In this study, was developed a semiempirical mathematical model for a TEV incorporated in a DX-SAHP utilizing R290. The research was carried out in an experimental way, conducted using a fully instrumented DX-SAHP. A series of experiments tested different operational modes for water heating, subject to varying solar radiation intensities. A uniform methodology collected operational parameters, resulting in 150 experiments. The semiempirical model creation involved combining basic equations from the literature describing TEV operation and formulating an equation using multiple linear regression based on experimentally evaluated parameters. These parameters included solar radiation incidence, inlet and outlet pressures, and superheating degree. The regression model achieved an adjustment higher than 96 %. Combining this regression with equations suggested by previous researchers, resulted in a model with an adjustment exceeding 94 %. Furthermore, this study culminated in the derivation of a generalized model primed for broad application across various heat pump configurations for water heating purposes. Notably, the model underwent rigorous validation across a separate set of experiments, reaffirming its reliability and versatility. While excelling in steady-state operations, the model's transient data analysis falls short, as intended. [ABSTRACT FROM AUTHOR]

As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm (Perinereis aibuhitensis) was expressed in E. coli (Escherichia coli) to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed in E. coli with low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein’s solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm in E. coli and broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease.

The aim of the present study is to trace the emergence and development of TEV and PVMV, new viruses for tobacco in Bulgaria, in the three varietal groups of Burley, Virginia and Basmi tobacco grown in the central part of southern Bulgaria. During the period 2017-2019, immunological studies were carried out to prove new viruses for tobacco in Bulgaria from the group of potyviruses PVMV and TEV. A kit for serological identification of the French company SEDIAG S.A.S. was used to prove the viruses. To detect TEV, TAS-ELISA was performed with IgG dilution of 1:100 and DAS-ELISA - to detect PVMV with IgG dilution of 1:100. In Virginia and Basmi tobacco variety groups, the potyvirus PVMV was serologically proven, and the virus was identified for the first time in tobacco in Bulgaria. PVMV was demonstrated in monoinfection and in complex with TEV in isolates from the cultivar group Virginia and Basmi. TEV and PVMV were not detected in isolates from the Burley cultivar group. In tobacco, the two potyviruses cannot be distinguished based on symptoms alone. Accurate diagnosis is possible by applying an ELISA or PCR test. [ABSTRACT FROM AUTHOR]

Fusion expression is widely employed to enhance the solubility of recombinant proteins. However, removal of the fusion tag is often required due to its potential impact on the structure and activity of passenger proteins. Tobacco etch virus (TEV) protease is widely used for this purpose due to its stringent sequence recognition. In the present work, fusion to the acyl carrier protein from E. coli fatty acid synthase (ACP) significantly increased the yield of recombinant soluble TEV, and the ACP tag also greatly improved TEV stability. The cleavage activity of TEV was not affected by the ACP fusion tag, and ACP-TEV retained high activity, even at unfavourable pH values. Moreover, ACP-TEV could be efficiently modified by co-expressed E. coli holo-ACP synthase (AcpS), leading to covalent attachment of 4′-phosphopantetheine (4′-PP) group to ACP. The sulfhydryl group of the long, flexible 4′-PP chain displayed high specific reactivity with iodoacetyl groups on the solid support. Thus, TEV could be immobilised effectively and conveniently via the active holo-ACP, and immobilised TEV retained high cleavage activity after a long storage period and several cycles of reuse. As a low-cost and recyclable biocatalyst, TEV immobilised by this method holds promise for biotechnological research and development. Key points: • The ACP tag greatly increased the soluble expression and stability of TEV protease. • The ACP tag did not affect the cleavage activity of TEV. • The holo-ACP Tag effectively mediated the covalent immobilisation of TEV. [ABSTRACT FROM AUTHOR]

A Disintegrin and Metalloprotease 10, also known as ADAM10, is a cell surface protease ubiquitously expressed in mammalian cells where it cuts several membrane proteins implicated in multiple physiological processes. The dysregulation of ADAM10 expression and function has been implicated in pathological conditions, including Alzheimer's disease (AD). Although it has been suggested that ADAM10 is expressed as a zymogen and the removal of the prodomain results in its activation, other potential mechanisms for the ADAM10 proteolytic function and activation remain unclear. Another suggested mechanism is post-translational modification of the cytoplasmic domain, which regulates ADAM10-dependent protein ectodomain shedding. Therefore, the precise and temporal activation of ADAM10 is highly desirable to reveal the fine details of ADAM10-mediated cleavage mechanisms and protease-dependent therapeutic applications. Here, we present a strategy to control prodomain and cytosolic tail cleavage to regulate ADAM10 shedding activity without the intervention of small endogenous molecule signaling pathways. We generated a series of engineered ADAM10 analogs containing Tobacco Etch Virus protease (TEV) cleavage site (TEVcs), rendering ADAM10 cleavable by TEV. This strategy revealed that, in the absence of other stimuli, the TEV-mediated removal of the prodomain could not activate ADAM10. However, the TEV-mediated cleavage of the cytosolic domain significantly increased ADAM10 activity. Then, we generated ADAM10 with a minimal constitutively catalytic activity that increased significantly in the presence of TEV or after activating a chemically activatable TEV. Our results revealed a bioengineering strategy for controlling the ADAM10 activity in living cells, paving the way to obtain spatiotemporal control of ADAM10. Finally, we proved that our approach of controlling ADAM10 promoted α-secretase activity and the non-amyloidogenic cleavage of amyloid-β precursor protein (APP), thereby increasing the production of the neuroprotective soluble ectodomain (sAPPα). Our bioengineering strategy has the potential to be exploited as a next-generation gene therapy for AD. [ABSTRACT FROM AUTHOR]

Socioemotional wealth (SEW) theory has a dominant role among family business theories due to its distinctive characteristics compared to non-family firms. Since 2007, the SEW concept has been widely used, and has examined the uniqueness of family firms. Meanwhile, critical voices have also emerged in the international academic discourse. The goal of this study is to offer a fresh literature review of SEW focusing on its changing priorities in multigenerational family businesses. This is a research area that is falsely neglected, considering the hereditary aspirations of family businesses and their consequences on preserving family control. Based upon the findings, propositions and further research directions are established. [ABSTRACT FROM AUTHOR]

The tobacco etch virus (TEV) protease is a commonly used reagent for removal of solubility and purification tags from recombinant proteins and is cited as being highly specific for its canonical cleavage site. Flexibility in some amino acids within this recognition sequence has been described in the literature but researchers generally assume few native human proteins will carry off-target sequences for TEV cleavage. We report here the aberrant cleavage of three human proteins with non-canonical TEV protease cleavage sites and identify broader sequence specificity rules that can be used to predict unwanted cleavage of recombinant proteins. Using these rules, 456 human proteins were identified that could be substrates for unwanted TEV protease cleavage. • Tobacco etch virus (TEV) protease has broader specificity than previously identified. • MOTIF searching can predict potential aberrant TEV protease cleavage sites in proteins. • TEV protease specificity involves both sequence and context dependent features. [ABSTRACT FROM AUTHOR]

The ISS-CREAM instrument is the modified version of the Cosmic Ray Energetics And Mass (CREAM) experiment, which was flown on balloons multiple times over Antarctica and later installed on the International Space Station (ISS). Its primary objective is to measure the energy spectra of individual cosmic-ray elements for the charge range of Z = 1 to Z = 26, in the energy range of ∼ 1012 to ∼ 1015 eV. The instrument comprises a tungsten/scintillator calorimeter and a pixelated silicon charge detector as primary detectors to determine the energy and charge of cosmic rays. Additionally, it includes top and bottom scintillator counting detectors and a boronated scintillator detector to differentiate between electrons and hadrons for multi-TeV electron measurements. The ISS-CREAM instrument was installed on the ISS in August 2017 and operated until February 2019. This paper provides an overview of the instrument, focusing on its detectors, trigger systems, common electronics, and power systems. The paper highlights the modifications made to these components to optimize their performance for ISS operations. [ABSTRACT FROM AUTHOR]

Engineered proteases are promising tools to address physiological and pathophysiological questions as well as to develop new therapeutic approaches. Here we introduce a new genetically encoded engineered single‐chain tobacco etch virus protease, allowing to control proprotein cleavage in different compartments of living mammalian cells. We demonstrated a set of controllable proteolytic effects, including cytosolic protein cleavage, inducible gene expression, and maturation of brain‐derived neurotrophic factor (BDNF) in the secretory pathway thus showing the versatility of this technique. Of note, the secretory pathway exhibits different characteristics from the cytosol and it is difficult to target because inaccessible to some small molecules. We were able to induce ligand‐mediated BDNF maturation and monitor its effects on dendritic spines in hippocampal pyramidal cells and in the mouse brain. This strategy paves the way to dissect proteolytic cleavage product signaling in various processes as well as for future therapeutic applications. [ABSTRACT FROM AUTHOR]

This research aims to boost tourism development in natural protected areas through the classification of the tourism hiking trails based on biodiversity ecosystem services values. The Total Economic Value (TEV) approach was used as the main research method to estimate the monetary value of the various ecosystem resources in Abu Qubies Syrian protected area. Five main tourism hiking trails in Abu Qubies were identified in order to be classified based on the economic value of their ecosystem resources. The study findings highlighted the importance of protected areas in enhancing tourism activities, especially natural-based ones. Additionally, this research identified the most economically valuable tourism hiking trails in the reserve, thus providing a supporting tool for decision-makers regarding tourism development in protected areas. This study presents the importance of the conservation perspective of natural resources in protected areas without ignoring their physical monetary value that can help governments in boosting local communities as well as the national, regional and local economies. [ABSTRACT FROM AUTHOR]

Abstract Engineered proteases are promising tools to address physiological and pathophysiological questions as well as to develop new therapeutic approaches. Here we introduce a new genetically encoded engineered single‐chain tobacco etch virus protease, allowing to control proprotein cleavage in different compartments of living mammalian cells. We demonstrated a set of controllable proteolytic effects, including cytosolic protein cleavage, inducible gene expression, and maturation of brain‐derived neurotrophic factor (BDNF) in the secretory pathway thus showing the versatility of this technique. Of note, the secretory pathway exhibits different characteristics from the cytosol and it is difficult to target because inaccessible to some small molecules. We were able to induce ligand‐mediated BDNF maturation and monitor its effects on dendritic spines in hippocampal pyramidal cells and in the mouse brain. This strategy paves the way to dissect proteolytic cleavage product signaling in various processes as well as for future therapeutic applications.

HC-Pro is a remarkable multifunctional protein of the genera Potyvirus and other viruses within the family Potiviridae. At least three main functions have been identified for this protein i) helper component of insect transmission; ii) RNA silencing suppressor and iii) viral polyprotein maturation. Evidence shows that this protein could be involved in other functions during the viral cycle. Dou to its activity HC-Pro is widely used in biotechnological studies related to different cellular process activated during plant-pathogen interaction. This protein is also employed in recombinant proteins production enhancement using plants as expression system. The aim of this work is to analyze the role reported in literature of HC-Pro from Tobacco etch virus as gene silencing suppressor in plants and to establish its possible applications in biotechnology. [ABSTRACT FROM AUTHOR]

Abstract Background Cell-free protein expression has become a widely used alternative of in vivo, cell-based systems in functional and structural studies of proteins. The wheat germ-based method outstands from the commercially available eukaryotic in vitro translation systems by its flexibility, high translation efficiency and success rate of properly folded eukaryotic protein synthesis. The original T7 promoter containing pEU3-NII vector was improved previously by addition of a ligation-independent cloning site, His6- and GST-tags, and a TEV protease cleavage site to facilitate the creation of recombinant plasmids, permit affinity purification, and enable production of purified, tag-free target proteins, respectively. Results Here, we describe a further development of pEU3-NII vector by inserting the rare-cutting, NotI restriction enzyme cleavage site to simplify vector linearization step prior to in vitro transcription. Additionally, His12, FLAG, and Halo affinity tag coding vectors have been created to increase detection sensitivity, specificity of interaction studies, and provide covalently linkable ligands for pull-down assays, respectively. Finally, the presented GST-His6, and GST-biotin double-tagging vectors could broaden the range of possibilities of protein-protein interaction studies. Conclusions The new generation of pEU3-NII vector family allows a more rapid production of translationally active mRNA and wheat germ cell-free expression of target proteins with a wide variety of affinity tags thus enables designing flexible and diverse experimental arrangement for in vitro studies of proteins.

The needle valve (NEV) has high potential for further using as the automatic expansion device in the refrigeration system. However, the experimental proof is required for further developing the automatic controller for operation. This paper proposes the feasibility study and performance map of the needle valve (NEV) using as the expansion device in a small vapour compression refrigerator (VCR). The relevant parameters produced by NEV are compared with those using capillary tube (CT) and thermostatic expansion valve (TEV) which are the conventional device. The system COP, superheated degree at evaporator outlet (SHDevap-out), subcooled degree at condenser outlet (SCDcon-out), mass flow rate, volumetric refrigeation capacity, compressor's specific work and refrigerant quality are considered for comparisons. Additionally, the economic assessment of using NEV compared with the other two devices is proposed. It is found that COP produced by NEV are similar to TEV which is better than CT. The ecomnomic assessment indicates that the NEV produces similar useful cooling energy per unit cost to that using TEV which is higher than CT. Overall results indicate that the NEV can efficiently be used as the expansion device in VCR which is alternative to the conventional expansion devices.

Tn916-like conjugative transposons carrying antibiotic resistance genes are found in a diverse range of bacteria. Orf14 within the conjugation module encodes a bifunctional cell wall hydrolase CwlT that consists of an N-terminal bacterial lysozyme domain (N-acetylmuramidase, bLysG) and a C-terminal NlpC/P60 domain (γ-d-glutamyl-l-diamino acid endopeptidase) and is expected to play an important role in the spread of the transposons. We determined the crystal structures of CwlT from two pathogens, Staphylococcus aureus Mu50 (SaCwlT) and Clostridium difficile 630 (CdCwlT). These structures reveal that NlpC/P60 and LysG domains are compact and conserved modules, connected by a short flexible linker. The LysG domain represents a novel family of widely distributed bacterial lysozymes. The overall structure and the active site of bLysG bear significant similarity to other members of the glycoside hydrolase family 23 (GH23), such as the g-type lysozyme (LysG) and Escherichia coli lytic transglycosylase MltE. The active site of bLysG contains a unique structural and sequence signature (DxxQSSES+S) that is important for coordinating a catalytic water. Molecular modeling suggests that the bLysG domain may recognize glycan in a similar manner to MltE. The C-terminal NlpC/P60 domain contains a conserved active site (Cys-His-His-Tyr) that appears to be specific to murein tetrapeptide. Access to the active site is likely regulated by isomerism of a side chain atop the catalytic cysteine, allowing substrate entry or product release (open state), or catalysis (closed state).

Wetlands provide critical services such as protection against flooding and wave action, water quality maintenance, habitat provision, food products, etc. Ashtamudi is the second-largest wetland ecosystem in Kerala state, and it provides numerous ecosystem services (ES). This study has analysed the economic value and current status of 11 important ES provided by this wetland. The considered services are fish production, clam production, shrimp/prawn larvae production, inland navigation, coir production, carbon sequestration and erosion prevention provided by mangroves, flood protection, cooling effect, tourism and future use-value. The total economic value of the wetland by considering the aforementioned services is 424 million US$ (in terms of 2017 International $ value), and every hundred square metres of the wetland provides an economic output of 820 US$ per year approximately. Even though the valuation of a few ES has been carried out in earlier studies, this particular study has incorporated more services in valuation and analysed the change in its value compared to previous years. This study also tried to analyse the status of 'wetland features' essential for the sustenance of ES, the spatial changes of the service providing habitats, and its impact on ES delivery. The study has been carried out in a GIS framework using inputs from various sources including remote sensing (RS) data. Landsat imageries and digital elevation model of the study area are used to understand the economic value and current status of certain services. Analysis, such as spatial change of service providing habitats, shoreline alterations, changes in NDVI of selected locations is carried out by utilizing the aforementioned RS data. Compared to the previous years, the value of supporting and regulating services is declining. At the same time, the value of certain provisioning and cultural services is improving. As the supporting and regulating services are the prerequisites for all other ES to sustain, the current trend indicates a collapse of the entire system shortly. While addressing this issue, more priority has to be given in restoring antecedent wetland condition, which will indeed lead to the improvement of supporting and regulating ES. Community-based institutional mechanisms have to be developed to ensure consistent monitoring and maintenance of the wetland. [ABSTRACT FROM AUTHOR]

This Letter reports on a first measurement of the inclusive W+jets cross section in proton-proton collisions at a centre-of-mass energy of 7 TeV at the LHC, with the ATLAS detector. Cross sections, in both the electron and muon decay modes of the W-boson, are presented as a function of jet multiplicity and of the transverse momentum of the leading and next-to-leading jets in the event. Measurements are also presented of the ratio of cross sections σ(W+≥n)/σ(W+≥n-1) for inclusive jet multiplicities n=1-4. The results, based on an integrated luminosity of 1.3 pb-1, have been corrected for all known detector effects and are quoted in a limited and well-defined range of jet and lepton kinematics. The measured cross sections are compared to particle-level predictions based on perturbative QCD. Next-to-leading order calculations, studied here for n≤2, are found in good agreement with the data. Leading-order multiparton event generators, normalized to the NNLO total cross section, describe the data well for all measured jet multiplicities. © 2011 CERN Elsevier B.V.

This Letter reports on a first measurement of the inclusive W+jets cross section in proton-proton collisions at a centre-of-mass energy of 7 TeV at the LHC, with the ATLAS detector. Cross sections, in both the electron and muon decay modes of the W-boson, are presented as a function of jet multiplicity and of the transverse momentum of the leading and next-to-leading jets in the event. Measurements are also presented of the ratio of cross sections σ(W+≥n)/σ(W+≥n-1) for inclusive jet multiplicities n=1-4. The results, based on an integrated luminosity of 1.3 pb-1, have been corrected for all known detector effects and are quoted in a limited and well-defined range of jet and lepton kinematics. The measured cross sections are compared to particle-level predictions based on perturbative QCD. Next-to-leading order calculations, studied here for n≤2, are found in good agreement with the data. Leading-order multiparton event generators, normalized to the NNLO total cross section, describe the data well for all measured jet multiplicities. © 2011 CERN Elsevier B.V.

Dijet angular distributions from the first LHC pp collisions at center-of-mass energy s=7 TeV have been measured with the ATLAS detector. The dataset used for this analysis represents an integrated luminosity of 3.1 pb-1. Dijet Χ distributions and centrality ratios have been measured up to dijet masses of 2.8 TeV, and found to be in good agreement with Standard Model predictions. Analysis of the λ distributions excludes quark contact interactions with a compositeness scale Χ below 3.4 TeV, at 95% confidence level, significantly exceeding previous limits. © 2010 CERN.

Dijet angular distributions from the first LHC pp collisions at center-of-mass energy s=7 TeV have been measured with the ATLAS detector. The dataset used for this analysis represents an integrated luminosity of 3.1 pb-1. Dijet Χ distributions and centrality ratios have been measured up to dijet masses of 2.8 TeV, and found to be in good agreement with Standard Model predictions. Analysis of the λ distributions excludes quark contact interactions with a compositeness scale Χ below 3.4 TeV, at 95% confidence level, significantly exceeding previous limits. © 2010 CERN.

Background: Cell-free protein expression has become a widely used alternative of in vivo, cell-based systems in functional and structural studies of proteins. The wheat germ-based method outstands from the commercially available eukaryotic in vitro translation systems by its flexibility, high translation efficiency and success rate of properly folded eukaryotic protein synthesis. The original T7 promoter containing pEU3-NII vector was improved previously by addition of a ligation-independent cloning site, His6- and GST-tags, and a TEV protease cleavage site to facilitate the creation of recombinant plasmids, permit affinity purification, and enable production of purified, tag-free target proteins, respectively. Results: Here, we describe a further development of pEU3-NII vector by inserting the rare-cutting, NotI restriction enzyme cleavage site to simplify vector linearization step prior to in vitro transcription. Additionally, His12, FLAG, and Halo affinity tag coding vectors have been created to increase detection sensitivity, specificity of interaction studies, and provide covalently linkable ligands for pull-down assays, respectively. Finally, the presented GST-His6, and GST-biotin double-tagging vectors could broaden the range of possibilities of protein-protein interaction studies. Conclusions: The new generation of pEU3-NII vector family allows a more rapid production of translationally active mRNA and wheat germ cell-free expression of target proteins with a wide variety of affinity tags thus enables designing flexible and diverse experimental arrangement for in vitro studies of proteins. [ABSTRACT FROM AUTHOR]

Single nucleotide polymorphisms (SNP) associated with Venous Thromboembolism (VTE) risk have been identified in European and American populations. Replicate SNPs associated with VTE in a Brazilian multicenter case-control study of the Southeast region. Patients with previous VTE assisted at the Outpatient Clinics of 3 centers of the Southeast Brazilian region were compared to normal controls of the same geographic region. We evaluated 29 SNPs associated with VTE risk in other populations, and 90 SNPs for stratification analysis of the population. Due to high admixture of Brazilian population and lack of previous studies, the calculation of the sample power was performed after genotyping. Sample size, allelic frequency and Hardy-Weinberg equilibrium were estimated. The association and odds ratio analyses were estimated by logistic regression and the results were adjusted for multiple tests using Bonferroni correction. The evaluation of the genetic structure similarity in the cases and controls was performed by AMOVA. 436 cases and 430 controls were included. It was demonstrated that this sample has a statistical power to detect a genetic association of 79.4%. AMOVA showed that the genetic variability between groups was 0.0% and 100% within each group. None of the SNPs showed association with VTE in our population. A Brazilian multicenter case-control study with adequate sample power, high genetic variability though no stratification between groups, showed no replication of SNPs associated with VTE. The high admixture of Brazilian population may be responsible for these results, emphasizing the influence of the population genetic structure in association studies. [ABSTRACT FROM AUTHOR]

The subject of study in the present work is the spread of TEV from the potvirus group in Virginia, Burley and Basmi tobacco varieties. The aim is to monitor the occurrence and development of TEV in these varieties grown in the central parts of southern Bulgaria and to analyze their resistance to this virus. During the period 2010 - 2019 immunological studies were carried out to prove the new viral tobacco disease in Bulgaria. A serological identification kit of the French company SEDIAG S.A.S. was used to demonstrate the virus. To establish TEV, a TAS-ELISA was performed at 1: 100 dilution of IgG. The virus was found to be sporadic. From the eight years of the study, TEV has been proven in four. The 2010-2011 period is characterized by a significant prevalence of TEV in Burley Group tobacco (14.3% and 12.7%, respectively). The virus is most strongly developed in 2019. It is evidenced in 60% of the tested samples from the Virginia variety. During the eight-year study period, only in 2010 TEV was found in isolates from the three tobacco varieties - Virginia, Burley and Basmi. As a result of the single-factor analysis of variance and Duncan's test, it was proven that tobacco from the Basmi variety was the most susceptible to infection with the TEV virus (0.165), followed by Virginia (0.136). Burley tobacco was found most resistant to the disease of TEV (0.126). [ABSTRACT FROM AUTHOR]