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The rapidly developing agro-industry generates huge amounts of lignocellulosic crop residues and animal manure worldwide. Although co-composting represents a promising and cost-effective method to treat various agricultural wastes simultaneously, poor composting efficiency prolongs total completion time and deteriorates the quality of the final product. However, supplementation of the feedstock with beneficial microorganisms can mitigate these negative effects by facilitating the decomposition of recalcitrant materials, enhancing microbial enzyme activity, and promoting maturation and humus formation during the composting process. Nevertheless, the influence of microbial inoculation may vary greatly depending on certain factors, such as start-up parameters, structure of the feedstock, time of inoculation, and composition of the microbial cultures used. The purpose of this contribution is to review recent developments in co-composting procedures involving different lignocellulosic crop residues and farm animal manure combined with microbial inoculation strategies. To evaluate the effectiveness of microbial additives, the results reported in a large number of peer-reviewed articles were compared in terms of composting process parameters (i.e., temperature, microbial activity, total organic carbon and nitrogen contents, decomposition rate of lignocellulose fractions, etc.) and compost characteristics (humification, C/N ratio, macronutrient content, and germination index). Most studies confirmed that the use of microbial amendments in the co-composting process is an efficient way to facilitate biodegradation and improve the sustainable management of agricultural wastes. Overall, this review paper provides insights into various inoculation techniques, identifies the limitations and current challenges of co-composting, especially with microbial inoculation, and recommends areas for further research in this field.
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Inoculantes Agrícolas , Compostagem , Animais , Animais Domésticos , Lignina , Esterco , Nitrogênio/análise , SoloRESUMO
BACKGROUND: The fusion of computer tomography and deep learning is an effective way of achieving improved image quality and artifact reduction in reconstructed images. OBJECTIVE: In this paper, we present two novel neural network architectures for tomographic reconstruction with reduced effects of beam hardening and electrical noise. METHODS: In the case of the proposed novel architectures, the image reconstruction step is located inside the neural networks, which allows the network to be trained by taking the mathematical model of the projections into account. This strong connection enables us to enhance the projection data and the reconstructed image together. We tested the two proposed models against three other methods on two datasets. The datasets contain physically correct simulated data, and they show strong signs of beam hardening and electrical noise. We also performed a numerical evaluation of the neural networks on the reconstructed images according to three error measurements and provided a scoring system of the methods derived from the three measures. RESULTS: The results showed the superiority of the novel architecture called TomoNet2. TomoNet2 improved the quality of the images according to the average Structural Similarity Index from 0.9372 to 0.9977 and 0.9519 to 0.9886 on the two data sets, when compared to the FBP method. This network also yielded the best results for 79.2 and 53.0 percent for the two datasets according to Peak-Signal-to-Noise-Ratio compared to the other improvement techniques. CONCLUSIONS: Our experimental results showed that the reconstruction step used in skip connections in deep neural networks improves the quality of the reconstructions. We are confident that our proposed method can be effectively applied to other datasets for tomographic purposes.
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Processamento de Imagem Assistida por Computador , Redes Neurais de Computação , Artefatos , Processamento de Imagem Assistida por Computador/métodos , Razão Sinal-Ruído , Tomografia Computadorizada por Raios XRESUMO
A retrotransposon insertion in the SILV gene is associated with a peculiar phenotype of dog, known as a merle. It is characterised by various areas of their coat colour becoming diluted due to a malfunction in the eumelanin-producing pigment cells. Recent studies have shown that the exact size of the short interspersed element (SINE) insertion is in correlation with specific phenotypic attributes, but was not able to absolutely confine dogs to a certain colour pattern. Our study focused on the merle variations occurring in the Mudi breed. Altogether, 123 dog samples from 11 countries were tested and genotyped. The exact length of the merle alleles were determined by automated fluorescent capillary fragment analysis. The most frequent merle genotype in this Mudi sample collection was the 'classic' merle (m/M: 61.8%), whereas other variants, such as atypical (m/Ma and m/Ma+: 5.7%), harlequin (m/Mh: 13.8%), double merle (M/M: 0.8%) and mosaic profiles (17.9%) were also observed. The practical significance of testing this mutation is that, phenotypically, not only merle dogs are carriers of this insertion, but also the so-called hidden merle individuals (where the merle phenotype is fully covered by the pheomelanin-dominated colouration) are potentially capable of producing unintentionally homozygous 'double merle' progeny with ophthalmologic, viability and auditory impairments.
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Cães/fisiologia , Variação Genética , Genótipo , Fenótipo , Pigmentação/genética , Antígeno gp100 de Melanoma/genética , Alelos , Animais , Cruzamento , Cães/genética , Homozigoto , Mutação , Antígeno gp100 de Melanoma/metabolismoRESUMO
INTRODUCTION: Regardless of the good outcomes of conservatively treated fingertip injuries, many patients complain about volume defects and hypoesthesia. Injection of autologous fat into the defect zone may resolve the volume problem and stimulate digital nerve regeneration by adipose derived stem cell transplantation. METHODS: We analyzed 5 volume defects resulting from conservatively treated fingertip injuries in 4 patients (male to female ratio, 2:2), 6 months after the injection of autologous abdominal fat into the defect zone retrospectively. The fat transplantation was performed after complete wound healing was achieved (2.5-13.5 months; mean, 6.5 months). Assessment of 2-point discrimination; finger pulp circumference; the Disability of the Arm, Shoulder, and Hand score; visual analog scale score; and a questionnaire regarding the treatment satisfaction were carried out both preoperatively and at a 6 month follow-up according to the standard of our clinic. RESULTS: In 3 patients, one session of lipofilling was sufficient to correct the contour, whereas 1 patient with 2 fingertip injuries was treated a second time. There was no significant change in finger pulp circumference, in the 2-point discrimination values and the Disability of the Arm, Shoulder, and Hand score results. There was a significant improvement in the visual analog scale scores in 4 of 5 cases, and all patients were very satisfied with the results of the procedure regarding the improvement in hand function in sports, at work and their daily routine. CONCLUSIONS: Lipofilling can be performed with little operative risks, might alleviate the patient's symptoms and reduce pain after conservative treatment of fingertip injuries.
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Tecido Adiposo/transplante , Traumatismos dos Dedos/cirurgia , Adulto , Avaliação da Deficiência , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Transplante Autólogo , CicatrizaçãoRESUMO
The Porcine Reproductive and Respiratory Syndrome (PRRS) eradication program in Hungary, implemented between 2014 and 2022, utilized complete herd replacement and the introduction of high-performance breeds to enhance production efficiency and environmental sustainability in the swine sector. As a result, the sow population was reduced by 26.2% while maintaining nearly the same number of slaughter pigs. This led to significant reductions in ammonia emissions (-145,857 kg), slurry production (-153,879 m3), nitrogen emissions (-1,409,951 kg), and overall greenhouse gas emissions (91,768,362 kg CO2eq). Additionally, the feed and water consumption were substantially decreased by 53,237,805 kg and 292,978,094 L, respectively, further lowering the sector's environmental footprint. The study demonstrates the effectiveness of customized eradication strategies and advanced breeding practices in reducing the environmental impact of animal husbandry. These findings underscore the necessity for ongoing collaboration among scientists, policymakers, and industry stakeholders to develop and implement sustainable livestock production methods. The Hungarian experience provides valuable insights into how targeted interventions can simultaneously improve production outcomes and reduce the environmental burden in the swine industry.
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In this study, our focus was on developing and investigating nanoparticle-filled polypropylene-based single polymer composite foams. These composites had porous and nanotube-reinforced matrices, with plain woven polypropylene (PP) fabric as reinforcement. Our main objective was to enhance the energy absorption and stiffness of the single polymer composites (SPCs) by modifying their matrices. We produced SPCs with two different matrices: one of amorphous poly-alpha-olefin (APAO) and one of thermoplastic elastomer (TPE) blended with APAO. We observed that the APAO matrix exhibited better impregnation of the fabric due to its low viscosity, while the composites with the TPE matrix showed significantly better tensile properties. The foaming process applied to the matrices resulted in a substantial increase in energy absorption for the SPCs, while preserving their tensile properties relative to their density. Scanning electron microscope images confirmed that foaming of the APAO matrix was notably more effective, primarily due to its low viscosity. Furthermore, we successfully enhanced the stiffness and tensile properties of the SPCs by nano-reinforcing the matrices with multi-wall carbon nanotubes (MWCNTs). Due to the size of the nanotubes, this reinforcement did not compromise the impact properties of the SPCs. Scanning electron microscope images also demonstrated improved dispersion of the nanotubes within the APAO matrices.
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Soil-borne phytopathogens can have detrimental effects on both cereal and horticultural crops resulting in serious losses worldwide. Due to their high efficiency and easy applicability, synthetic pesticides are still the primary choice in modern plant disease control systems, but stringent regulations and increasing environmental concerns make the search for sustainable alternatives more pressing than ever. In addition to the incorporation of botanicals into agricultural practices, the diversification of cropping systems with aromatic and medicinal plants is also an effective tool to control plant diseases through providing nutrients and shaping soil microbial communities. However, these techniques are not universally accepted and may negatively affect soil fertility if their application is not thoroughly controlled. Because the biocontrol potential of aromatic and medicinal plants has been extensively examined over the past decades, the present study aims to overview the recent literature concerning the biopesticide effect of secondary metabolites derived from aromatic and medicinal plants on important soil-borne plant pathogens including bacteria, fungi, and nematodes. Most of the investigated herbs belong to the family of Lamiaceae (e.g., Origanum spp., Salvia spp., Thymus spp., Mentha spp., etc.) and have been associated with potent antimicrobial activity, primarily due to their chemical constituents. The most frequently tested organisms include fungi, such as Rhizoctonia spp., Fusarium spp., and Phytophthora spp., which may be highly persistent in soil. Despite the intense research efforts dedicated to the development of plant-based pesticides, only a few species of aromatic herbs are utilized for the production of commercial formulations due to inconsistent efficiency, lack of field verification, costs, and prolonged authorization requirements. However, recycling the wastes from aromatic and medicinal plant-utilizing industries may offer an economically feasible way to improve soil health and reduce environmental burdens at the same time. Overall, this review provides comprehensive knowledge on the efficiency of aromatic herb-based plant protection techniques, and it also highlights the importance of exploiting the residues generated by aromatic plant-utilizing sectors as part of agro-industrial processes.
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In this study, we used four amorphous poly-alpha-olefin (APAO) grades to improve the toughness of drawn polypropylene (PP) tapes. The samples containing different amounts of APAOs were drawn in a heat chamber of a tensile testing machine. The APAOs reduced the work of drawing and increased the melting enthalpy of the drawn specimens, as they facilitated the movement of the PP molecules. The APAO with the highest molecular weight and with a low level of crystallinity increased both the tensile strength and the strain-at-break of the specimens, so we also produced drawn tapes from that PP/APAO blend on a continuous-operation stretching line. The continuously drawn tapes also showed improved toughness.
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The golden jackal (Canis aureus) is a reoccurring species in the centre of the Carpathian basin, in Hungary. In total, 31 golden jackal tissue samples were collected, from 8 white-coated, 2 black-coated and one mottled animal across Hungary. Sequences and fragment length polymorphisms were studied for white colour (MC1R), and for black coat colouration (CBD103). In each white animal, the most widespread mutation causing white fur colour in dogs in homozygous form was detected. Three animals were found to carry the mutation in heterozygous form. The two black golden jackals were heterozygous for the 3 bp deletion in CBD103 that mutation for black coat colouration in dogs, and one of them also carried the mutation causing white fur. None of the white animals showed signs of hybridization, but both the black and the mottled coloured individuals were found to be hybrids based on genetic testing. Kinship was found three times, twice between white animals, and once between a white animal and an agouti animal carrying the mutation of white coat. Our results confirm the findings that golden jackal-dog hybrids may occur without human intervention, and the detected mutation causing white fur colour in golden jackals could possibly be due to an early hybridization event.
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Canidae , Chacais , Humanos , Cães , Animais , Chacais/genética , Mutação , Hibridização Genética , HungriaRESUMO
The aim of this study was to determine the breed boundary of the Hungarian Short-haired Vizsla (HSV) dog breed. Seventy registered purebred HSV dogs were genotyped on approximately 145,000 SNPs. Principal Component Analysis (PCA) and Admixture analysis certified that they belong to the same population. The outer point of the breed demarcation was a single Hungarian Wire-haired Vizsla (HWV) individual, which was the closest animal genetically to the HSV population in the PCA analysis. Three programs were used for the breed assignment calculations, including the widely used GeneClass2.0 software and two additional approaches developed here: the 'PCA-distance' and 'IBS-central' methods. Both new methods calculate a single number that represents how closely a dog fits into the actual reference population. The former approach calculates this number based on the PCA distances from the median of HSV animals. The latter calculates it from identity by state (IBS) data, measuring the distance from a central animal that is the best representative of the breed. Having no mixed-breed dogs with known HSV genome proportion, admixture animals were simulated by using data of HSV and HWV individuals to calibrate the inclusion/exclusion probabilities for the assignment. The numbers generated from these relatively simple calculations can be used by breeders and clubs to keep their populations under genetic supervision.
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Cães , Polimorfismo de Nucleotídeo Único , Animais , Cães/genética , Genoma , Genótipo , HungriaRESUMO
The quality of chemically bonded sand cores used during the manufacturing process of cast components is highly dependent on the properties of the sand, which constitutes the refractory base media of the core. One of the main advantages of the application of different types of sands as molding aggregates that after casting, they can be reclaimed and can be used again during core shooting. The properties of the sand, however, could be remarkably changed during the casting and reclamation processes. This study aims to investigate the effects of the properties of the base sand on the mechanical strength and thermal distortion properties of samples made from new and thermally reclaimed silica sand. For this purpose, particle size analysis, specific surface area, and loss on ignition measurements, as well as differential thermal analysis coupled with thermogravimetry, were executed on the base sands, and the sand grains were analyzed with scanning electron microscopy and X-ray diffraction. Test pieces were made with hot box and cold box technology for bending and hot distortion tests. It was found that by the utilization of reclaimed sand, cores with higher average bending strength and lower thermal deformation can be produced. These differences can be traced back to the more advantageous granulometric properties, lower impurity content, and lower thermal expansion of thermally reclaimed sand.
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Due to the steadily increasing need for seafood and the plateauing output of fisheries, more fish need to be produced by aquaculture production. In parallel with the improvement of farming methods, elite food fish lines with superior traits for production must be generated by selection programs that utilize cutting-edge tools of genomics. The purpose of this review is to provide a historical overview and status report of a selection program performed on a catadromous predator, the Asian seabass (Lates calcarifer, Bloch 1790) that can change its sex during its lifetime. We describe the practices of wet lab, farm and lab in detail by focusing onto the foundations and achievements of the program. In addition to the approaches used for selection, our review also provides an inventory of genetic/genomic platforms and technologies developed to (i) provide current and future support for the selection process; and (ii) improve our understanding of the biology of the species. Approaches used for the improvement of terrestrial farm animals are used as examples and references, as those processes are far ahead of the ones used in aquaculture and thus they might help those working on fish to select the best possible options and avoid potential pitfalls.
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The primary purpose of this study was to investigate the influence of Cellulomonas flavigena and Streptomyces viridosporus, as a bacterial inoculant, on the compostability of post-extraction lavender waste. The major physicochemical, microbiological, and biological properties of the composting materials were monitored for 161 days. The technology developed was shown to improve the compostability of recalcitrant herbal residues. The use of lavender waste beneficially affected the composting process by extending the thermophilic phase, accelerating the degradation of organic matter, and elevating the viable counts of useful microorganisms; however, adverse effects were also observed, including an increased carbon-to-nitrogen ratio (19.05) and a decreased germination index (93.4%). Bacterial inoculation was found to preserve the nitrogen content (2.50%) and improve the efficiency of biodegradation. The Salmonella- and Escherichia coli-free final composting products were mature, stable, and ready for soil application. To the authors' knowledge, no previous research has investigated the compostability of lavender waste. Likewise, this is the first study that has used strains of C. flavigena and S. viridosporus in combination to facilitate a composting process.
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The role of the surgical intervention is decisive in treating colorectal tumors. The neo-adjuvant radio-chemotherapy has improved the efficacy of the treatment of advanced rectum tumors. In order to decrease the size and stage of advanced rectal carcinoma and to increase the rate of resecability, we introduced neoadjuvant radio-chemotherapy. We carried out neo-adjuvant and surgical treatment in case of 67 patients with rectal adenocarcinoma (T 2-4 N 1-2 M 0 ) between June 1, 2005 and July 31, 2008. The average age of the patients was 61.2 years, the division according to sex was 44 males/23 females. Regarding the local stage of the rectal process or the proximity to the sphincter, we applied radio-chemotherapy (radiotherapy 25 times altogether 45 Gy and on the first and last week for 5-5 days they received 350 mg/m 2 /day 5-FU and 20 mg/m 2 /day leucovorin chemotherapy, recently complemented with 3 x 1.8 Gy advanced boost radiation aiming at the macroscopic tumor site with security zone). Patients underwent surgery 8 weeks on average after restaging examinations. Thirty-eight patients underwent anterior rectal resection with double stapler procedure; there were 18 abdominoperineal rectal extirpations, 7 Hartmann operations and 4 per annum excisions. Compared to the preoperative staging, the histological evaluation of the resected specimens showed total remission (pT 0 N 0 ) in 11% and partial remission in 43%. The morbidity necessitating reoperation was 5.9%, without mortality and suture insufficiency. The long-term neo-adjuvant oncological treatment led to down-staging of rectal tumors in most cases and increased the resecability and rate of resection operations.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais/sangue , Procedimentos Cirúrgicos do Sistema Digestório/métodos , Fracionamento da Dose de Radiação , Terapia Neoadjuvante/métodos , Neoplasias Retais/patologia , Neoplasias Retais/terapia , Idoso , Idoso de 80 Anos ou mais , Quimioterapia Adjuvante , Feminino , Humanos , Hungria , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Radioterapia Adjuvante , Neoplasias Retais/sangue , Estudos Retrospectivos , Fatores de Tempo , Resultado do TratamentoRESUMO
We developed polypropylene-based single-polymer composites (PP-SPC) with blends of amorphous poly-alpha-olefin (APAO) and random polypropylene copolymer (rPP) as matrix material and polypropylene (PP) woven fabric as reinforcement. Our goal was to utilize the lower melting temperature of APAO/rPP blends to increase the consolidation of the composites and decrease the heat load of the PP reinforcement. We produced the composites by film-stacking at 160 °C, and characterized the composites with density, peel, static tensile and dynamic falling weight impact tests, and by scanning electron microscopy. The results indicate that consolidation can be enhanced by increasing the APAO content of the matrix. We found that the APAO content of 50% is optimal for tensile properties. With increasing APAO content, the perforation energy decreased, but even the well-consolidated composites showed very high perforation energy. In the case of a pure APAO matrix, fiber content can be increased up to 80 wt% without a severe loss of consolidation, resulting in good tensile properties. The PP-SPCs developed possessed excellent mechanical properties, and well-consolidated composites can be produced with APAO/rPP blends as a matrix with high fiber content.
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The primary purpose of this research was to examine the bacteriological properties of raw ovine milk produced by Merino, Tsigai, Dorper, Lacaune, and British Milk Sheep flocks on four sheep farms located in the eastern part of Hungary. In addition to individual raw milk (IRM) and bulk tank milk (BTM) samples, the udder surface (US) of ewes was also tested for bacteriological quality. A total of 77 US, 77 IRM, and 10 BTM samples were collected in the early morning during regular milking sessions. The samples, kept cooled at temperatures below 4 °C, were delivered to the microbiological laboratory and were examined immediately. The relatively low numbers of bacteria in both US and IRM samples reflected good housing conditions of ewes kept on the four farms studied. However, BTM samples had up to 3.5-4.0 log10 CFU/mL higher mean bacterial counts than their IRM counterparts, and the mean levels of bacteria in BTM on two farms even exceeded the regulatory limit of 6.18 log10 CFU/mL. Further studies need to be performed to clarify this issue.
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Substitution of terpyridine at the 4' position with electron withdrawing and donating groups is used to tune the quintet lifetime of its iron(ii) complex. DFT calculations suggest that the energy barrier between the quintet and singlet states can be altered significantly upon substitution, inducing a large variation of the lifetime of the photoexcited quintet state. This prediction was experimentally verified by transient optical absorption spectroscopy and good agreement with the trend expected from the calculations was found. This demonstrates that the potential energy landscape can indeed be rationally tailored by relevant modifications based on DFT predictions. This result should pave the way to advancing efficient theory-based ligand engineering of functional molecules to a wide range of applications.
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The intensity of the merle pattern is determined by the length of the poly(A) tail of a repeat element which has been inserted into the boundary of intron 10 and exon 11 of the PMEL17 locus in reverse orientation. This poly(A) tail behaves as a microsatellite, and due to replication slippage, longer and shorter alleles of it might be generated during cell divisions. The length of the poly(A) tail regulates the splicing mechanism. In the case of shorter tails, the removal of intron 10 takes place at the original splicing, resulting in a normal premelanosome protein (PMEL). Longer tails generate larger insertions, forcing splicing to a cryptic splice site, thereby coding for an abnormal PMEL protein, which is unable to form the normal fibrillar matrix of the eumelanosomes. Thus, eumelanin deposition ensuring the dark color formation is reduced. In summary, the longer the poly(A) tail, the lighter the coat color intensity of the melanocytes. These mutations can occur in the somatic cells and the resulting cell clones will shape the merle pattern of the coat. When they take place in the germ line, they occasionally produce offspring with unexpected color variations which are different from those of their parents.
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Cor de Cabelo/genética , Splicing de RNA/genética , Antígeno gp100 de Melanoma/genética , Animais , Cães , Patrimônio Genético , Melaninas/biossíntese , Melaninas/genética , Melanócitos/metabolismo , Mutação/genética , Sítios de Splice de RNA/genéticaRESUMO
Kaposi's sarcoma (KS), a vascular tumor associated with human immunodeficiency virus type 1 infection, is characterized by spindle-shaped endothelial cells, inflammatory cells, cytokines, growth and angiogenic factors, and angiogenesis. KS spindle cells are believed to be of the lymphatic endothelial cell (LEC) type. Kaposi's sarcoma-associated herpesvirus (KSHV, or human herpesvirus 8) is etiologically linked to KS, and in vitro KSHV infection of primary human dermal microvascular endothelial cells (HMVEC-d) is characterized by the induction of preexisting host signal cascades, sustained expression of latency-associated genes, transient expression of a limited number of lytic genes, sustained induction of NF-kappaB and several cytokines, and growth and angiogenic factors. KSHV induced robust vascular endothelial growth factor A (VEGF-A) and VEGF-C gene expression as early as 30 min postinfection (p.i.) in serum-starved HMVEC-d, which was sustained throughout the observation period of 72 h p.i. Significant amounts of VEGF-A and -C were also detected in the culture supernatant of infected cells. VEGF-A and -C were also induced by UV-inactivated KSHV and envelope glycoprotein gpK8.1A, thus suggesting a role for virus entry stages in the early induction of VEGF and requirement of KSHV viral gene expression for sustained induction. Exogenous addition of VEGF-A and -C increased KSHV DNA entry into target cells and moderately increased latent ORF73 and lytic ORF50 promoter activation and gene expression. KSHV infection also induced the expression of lymphatic markers Prox-1 and podoplanin as early as 8 h p.i., and a paracrine effect was seen in the neighboring uninfected cells. Similar observations were also made in the pure blood endothelial cell (BEC)-TIME cells, thus suggesting that commitment to the LEC phenotype is induced early during KSHV infection of blood endothelial cells. Treatment with VEGF-C alone also induced Prox-1 expression in the BEC-TIME cells. Collectively, these studies show that the in vitro microenvironments of KSHV-infected endothelial cells are enriched, with VEGF-A and -C molecules playing key roles in KSHV biology, such as increased infection and gene expression, as well as in angiogenesis and lymphangiogenesis, thus recapitulating the microenvironment of early KS lesions.
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Endotélio Vascular/virologia , Herpesvirus Humano 8/crescimento & desenvolvimento , Pele/virologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator C de Crescimento do Endotélio Vascular/metabolismo , Capilares , Linhagem Celular , Endotélio Vascular/metabolismo , Expressão Gênica/efeitos dos fármacos , Regulação Viral da Expressão Gênica , Herpesvirus Humano 8/efeitos dos fármacos , Herpesvirus Humano 8/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Glicoproteínas de Membrana/metabolismo , Regiões Promotoras Genéticas/efeitos dos fármacos , Pele/irrigação sanguínea , Pele/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/farmacologia , Fator C de Crescimento do Endotélio Vascular/genética , Fator C de Crescimento do Endotélio Vascular/farmacologiaRESUMO
Kaposi's sarcoma-associated herpesvirus (KSHV) interacts with cell surface heparan sulfate (HS) and alpha3beta1 integrin during the early stages of infection of human dermal microvascular endothelial cells (HMVEC-d) and human foreskin fibroblasts (HFF), and these interactions are followed by virus entry overlapping with the induction of preexisting host cell signal pathways. KSHV also utilizes the amino acid transporter protein xCT for infection of adherent cells, and the xCT molecule is part of the cell surface heterodimeric membrane glycoprotein CD98 (4F2 antigen) complex known to interact with alpha3beta1 and alphaVbeta3 integrins. KSHV gB mediates adhesion of HMVEC-d, CV-1, and HT-1080 cells and HFF via its RGD sequence. Anti-alphaV and -beta1 integrin antibodies inhibited the cell adhesion mediated by KSHV-gB. Variable levels of neutralization of HMVEC-d and HFF infection were observed with antibodies against alphaVbeta3 and alphaVbeta5 integrins. Similarly, variable levels of inhibition of virus entry into adherent HMVEC-d, 293 and Vero cells, and HFF was observed by preincubating virus with soluble alpha3beta1, alphaVbeta3, and alphaVbeta5 integrins, and cumulative inhibition was observed with a combination of integrins. We were unable to infect HT1080 cells. Virus binding and DNA internalization studies suggest that alphaVbeta3 and alphaVbeta5 integrins also play roles in KSHV entry. We observed time-dependent temporal KSHV interactions with HMVEC-d integrins and CD98/xCT with three different patterns of association and dissociation. Integrin alphaVbeta5 interaction with CD98/xCT predominantly occurred by 1 min postinfection (p.i.) and dissociated at 10 min p.i., whereas alpha3beta1-CD98/xCT interaction was maximal at 10 min p.i. and dissociated at 30 min p.i., and alphaVbeta3-CD98/xCT interaction was maximal at 10 min p.i. and remained at the observed 30 min p.i. Fluorescence microscopy also showed a similar time-dependent interaction of alphaVbeta5-CD98. Confocal-microscopy studies confirmed the association of CD98/xCT with alpha3beta1 and KSHV. Preincubation of KSHV with soluble heparin and alpha3beta1 significantly inhibited this association, suggesting that the first contact with HS and integrin is an essential element in subsequent CD98-xCT interactions. Anti-CD98 and xCT antibodies did not block virus binding and entry and nuclear delivery of viral DNA; however, viral-gene expression was significantly inhibited, suggesting that CD98-xCT play roles in the post-entry stage of infection, possibly in mediating signal cascades essential for viral-gene expression. Together, these studies suggest that KSHV interacts with functionally related integrins (alphaVbeta3, alpha3beta1, and alphaVbeta5) and CD98/xCT molecules in a temporal fashion to form a multimolecular complex during the early stages of endothelial cell infection, probably mediating multiple roles in entry, signal transduction, and viral-gene expression.