RESUMO
In recent years, anaerobic digestion has been recognized as a suitable alternative for tequila vinasses treatment due to its high energy recovery and chemical oxygen demand (COD) removal efficiency. However, key factors such as the lack of suitable monitoring schemes and the presence of load disturbances, which may induce unstable operating conditions in continuous systems, have limited its application at full scale. Therefore, the aim of this work was to evaluate the anaerobic sequencing batch reactor (AnSBR) configuration in order to provide a low cost and easy operation alternative for the treatment of these complex effluents. In particular, the AnSBR was evaluated under different pH-temperature combinations: 7 and 32 °C; 7 and 38 °C; 8 and 32 °C and 8 and 38 °C. Results showed that the AnSBR configuration was able to achieve high COD removal efficiencies (around 85%) for all the tested conditions, while the highest methane yield was obtained at pH 7 and 38 °C (0.29 L/g COD added). Furthermore, high robustness was found in all the AnSBR experiments. Therefore, the full-scale application of the AnSBR technology for the treatment of tequila vinasses is quite encouraging, in particular for small and medium size tequila industries that operate under seasonal conditions.
Assuntos
Biocombustíveis/análise , Resíduos Industriais/análise , Eliminação de Resíduos Líquidos , Bebidas Alcoólicas/análise , Anaerobiose , Reatores Biológicos , Concentração de Íons de Hidrogênio , Eliminação de Resíduos , TemperaturaRESUMO
As Artificial Intelligence (AI) proliferates across important social institutions, many of the most powerful AI systems available are difficult to interpret for end-users and engineers alike. Here, we sought to characterize public attitudes towards AI interpretability. Across seven studies (N = 2475), we demonstrate robust and positive attitudes towards interpretable AI among non-experts that generalize across a variety of real-world applications and follow predictable patterns. Participants value interpretability positively across different levels of AI autonomy and accuracy, and rate interpretability as more important for AI decisions involving high stakes and scarce resources. Crucially, when AI interpretability trades off against AI accuracy, participants prioritize accuracy over interpretability under the same conditions driving positive attitudes towards interpretability in the first place: amidst high stakes and scarce resources. These attitudes could drive a proliferation of AI systems making high-impact ethical decisions that are difficult to explain and understand.
Assuntos
Inteligência Artificial , Opinião Pública , Atitude , HumanosRESUMO
In sulfate-reducing reactors, it has been reported that the sulfate removal efficiency increases when the COD/SO4(2-) ratio is increased. The start-up of a down-flow fluidized bed reactor constitutes an important step to establish a microbial community in the biofilm able to survive under the operational bioreactor conditions in order to achieve effective removal of both sulfate and organic matter. In this work the influence of COD/SO4(2-) ratio and HRT in the development of a biofilm during reactor start-up (35 days) was studied. The reactor was inoculated with 1.6 g VSS/L of granular sludge, ground low density polyethylene was used as support material; the feed consisted of mineral medium at pH 5.5 containing 1 g COD/L (acetate:lactate, 70:30) and sodium sulfate. Four experiments were conducted at HRT of 1 or 2 days and COD/SO4(2-) ratio of 0.67 or 2.5. The results obtained indicated that a COD/SO4(2-) ratio of 2.5 and HRT 2 days allowed high sulfate and COD removal (66.1 and 69.8%, respectively), whereas maximum amount of attached biomass (1.9 g SVI/L support) and highest sulfate reducing biofilm activity (10.1 g COD-H2S/g VSS-d) was achieved at HRT of 1 day and at COD/sulfate ratios of 0.67 and 2.5, respectively, which suggests that suspended biomass also played a key role in the performance of the reactors.
Assuntos
Biofilmes , Oxigênio/química , Sulfatos/química , Reatores BiológicosRESUMO
Primitive neuroectodermal tumor (PNET) is part of the Ewing sarcoma family of tumors. The present case reports a primitive neuroectodermal tumor (PNET) of rare location in the bladder in a newborn. It was evaluated with prenatal ultrasound and postnatal tomography that revealed a mass in the posterior wall of the bladder. The patient underwent partial cystectomy with subsequent analysis of the surgical piece removed, the histopathological study indicated a tumor of mesenchymal origin, and immunohistochemical staining confirmed the diagnosis of PNET of the bladder. Satisfactory result and short-term follow-up.
RESUMO
Hepatitis B virus (HBV) antiviral drug resistance mutations prevent successful outcome of treatment and lead to worsening of liver disease. Detection of its emergence permits opportune treatment with alternative drugs. Unfortunately, the use of newly approved antivirals, including adefovir dipivoxil, emtricitabine, and telbivudine, is also associated with the development of drug resistance, albeit to a lesser extent than the use of lamivudine. The objectives of this work were to assess the performance characteristics (sensitivity and accuracy) of an updated drug resistance test, the INNO-LiPA HBV DR v2, which includes detection of mutations associated with lamivudine, adefovir, emtricitabine, and telbivudine resistance, and to compare the results with consensus sequencing of serum samples from patients treated with HBV antivirals. Diagnostic sensitivity, defined as detection of a positive amplification line on the line probe assay (LiPA) strip, was 94.8% (95% confidence interval [CI], 89.7 to 97.9) after initial testing, increasing to 96.3% (95% CI, 91.6 to 98.8) after repeat test 1 and to 100% (95% CI, 97.3 to 100.0) after repeat test 2. In diagnostic accuracy determinations, full concordance was observed between sequencing and LiPA for 77.0% of the codons tested (620/805 codons [95% CI, 74.0 to 79.9]), whereas LiPA and sequencing were partially concordant 22% of the time (177/805 codons). In 167 out of 177 cases, LiPA detected a wild-type/mutant mixture whereas sequencing detected only one of the two results. Performance testing of the new LiPA test, the INNO-LiPA HBV DR v2, showed convincing diagnostic sensitivity and accuracy. The ability of the test to detect mixed infections and minority viral populations associated with resistance to the current generation of antivirals, including adefovir, emtricitabine, and telbivudine, makes it a useful tool for HBV therapy monitoring.
Assuntos
Antivirais/farmacologia , Monitoramento de Medicamentos/métodos , Farmacorresistência Viral/genética , Vírus da Hepatite B/efeitos dos fármacos , Hibridização de Ácido Nucleico/métodos , Nucleosídeos/farmacologia , Antivirais/química , Antivirais/uso terapêutico , DNA Viral/sangue , Hepatite B/tratamento farmacológico , Hepatite B/virologia , Vírus da Hepatite B/genética , Humanos , Testes de Sensibilidade Microbiana , Mutação , Nucleosídeos/química , Nucleosídeos/uso terapêutico , Kit de Reagentes para Diagnóstico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNARESUMO
Detecting where an epidemic started, i.e., which node in a network was the source, is of crucial importance in many contexts. However, finding the source of an epidemic can be challenging, especially because the information available is often sparse and noisy. We consider a setting in which we want to localize the source based exclusively on the information provided by a small number of observers - i.e., nodes that can reveal if and when they are infected - and we study where such observers should be placed. We show that the optimal observer placement depends not only on the topology of the network, but also on the variance of the node-to-node transmission delays. We consider both low-variance and high-variance regimes for the transmission delays and propose algorithms for observer placement in both cases. In the low-variance regime, it suffices to only consider the network-topology and to choose observers that, based on their distances to all other nodes in the network, can distinguish among possible sources. However, the high-variance regime requires a new approach in order to guarantee that the observed infection times are sufficiently informative about the location of the source and do not get masked by the noise in the transmission delays; this is accomplished by additionally ensuring that the observers are not placed too far apart. We validate our approaches with simulations on three real-world networks. Compared to state-of-the-art strategies for observer placement, our methods have a better performance in terms of source-localization accuracy for both the low- and the high-variance regimes.
RESUMO
Ligands specific for B7.1 (CD80) and B7.2 (CD86) have applications in disease indications that require inhibition of T-cell activity. As we observed significant sequence and structural similarity between the B7-binding ligand, cytotoxic T-lymphocyte associated protein-4 (CTLA-4), and antibody variable light chain domains (VLs), we have explored the possibilities of making novel B7 binding molecules based on single VL domains. We first describe the "rational" design and construction of a VL/CTLA-4 hybrid molecule in which we have grafted both the CDR1 and CDR3-like loops of CTLA-4 onto a single VL light chain, at sites determined by sequence and structure-based alignment. This molecule was secreted as a soluble product from Escherichia coli, but did not show any binding to B7.1 and B7.2. In a second approach we constructed a VL library in which human VL genes derived from B-cells were spiked with the CDR3-like loop of CTLA-4 and further diversified by DNA shuffling. This library was displayed on phage, and after selection gave B7.1 binding ligands which competed with CTLA-4. In order to evaluate the possible general utility of VL domains as binding ligands, we have constructed a non-biased VL library. From this DNA-shuffled human VL library we have selected single VL domains specific for B7.1, B7.2 or human IgG. Two B7.1-specific VL ligands and one B7.2-specific VL ligand showed competition with CTLA-4. One candidate VL domain-specific for B7.1 was affinity matured by simultaneous randomisation of all CDR loops using DNA shuffling with degenerate CDR-spiking oligonucleotides. From this library, a single VL domain with affinity of 191 nM for B7.1 was obtained, which also showed binding to B7.1 in situ. This VL had mutations in CDR1 and CDR3, indicating that antigen recognition for this single VL is most likely mediated by the same regions as in the VL domain of whole antibodies. The B7.1 and B7.2-specific VL domains described in this study may form the basis of a new family of immunomodulatory recombinant molecules. Furthermore, our studies suggest that it is feasible to create specific single VL domains to diverse targets as is the case for single VH domains.
Assuntos
Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Evolução Molecular Direcionada/métodos , Cadeias Leves de Imunoglobulina/química , Região Variável de Imunoglobulina/química , Glicoproteínas de Membrana/metabolismo , Biblioteca de Peptídeos , Sequência de Aminoácidos , Afinidade de Anticorpos , Especificidade de Anticorpos , Antígenos CD/imunologia , Antígeno B7-1/imunologia , Antígeno B7-2 , Cromatografia em Gel , Clonagem Molecular , Regiões Determinantes de Complementaridade/química , Regiões Determinantes de Complementaridade/imunologia , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Cadeias Leves de Imunoglobulina/genética , Cadeias Leves de Imunoglobulina/imunologia , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Ligantes , Glicoproteínas de Membrana/imunologia , Dados de Sequência Molecular , Mutação/genética , Estrutura Terciária de Proteína , Análise de Sequência de DNA , Especificidade por Substrato , Ressonância de Plasmônio de SuperfícieRESUMO
RESUMEN El higroma quístico es la malformación del sistema linfático que más frecuentemente se observa en el período prenatal y que se ubica principalmente en el cuello y/o la nuca. Su tasa de detección ha aumentado desde la implementación de la translucencia nucal fetal (TN) en el primer trimestre de embarazo, y su presencia se ha relacionado con anomalías congénitas, aneuploidías, pérdida del embarazo y trastornos en el desarrollo. El objetivo de la presentación de este caso es resaltar la importancia del diagnóstico antenatal del higroma quístico, con el fin de realizar una intervención precoz y evitar la muerte fetal. Se recibe para estudio anatomopatológico, feto de sexo indeterminado producto del primer embarazo de una madre de 19 años de edad sin previos controles prenatales, con presencia de una gran masa quística que se extiende desde el rostro hasta la nuca. Mediante el estudio histológico se confirma el diagnóstico de higroma quístico. Al carecer de análisis de cariotipo no fue posible establecer la preexistencia de alguna anomalía genética. El también conocido como linfangioma quístico, es un tumor vascular benigno cuyo diagnóstico antenatal mediante la ultrasonografía resulta fundamental en la evolución y pronóstico de la enfermedad. Desafortunadamente en nuestro caso, la falta de controles prenatales y la ausencia de estudios ultrasonográficos que permitieran conocer las características de este linfangioma, pudo impactar significativamente en el desenlace fatal.
ABSTRACT The cystic hygroma is the malformation of the lymphatic system that is most frequently observed in the prenatal period and is located mainly in the neck and/or the nape of the neck. Its detection rate has increased since the implementation of fetal nuchal translucency (NT) in the first trimester of pregnancy and its presence has been associated with congenital abnormalities, aneuploidies, pregnancy loss, and developmental disorders. The aim of this case is to highlight the importance of antenatal diagnosis of cystic hygroma in order to perform early intervention and avoid fetal death. It is received, for anatomopathological study, a fetus of undetermined sex product of the first pregnancy of a 19 year-old mother without previous prenatal controls, with the presence of a large cystic mass that extends from the face to the neck. The histological study confirms the diagnosis of cystic hygroma. As there was no karyotype analysis, it was not possible to establish the preexistence of any genetic abnormality. Also known as cystic lymphangioma, is a benign vascular tumor whose antenatal diagnosis by ultrasonography is essential in the evolution and prognosis of the disease. Unfortunately in our case, the lack of prenatal controls and the absence of ultrasonographic studies that would allow knowing the characteristics of this lymphangioma, could significantly impact in the fatal outcome.
RESUMO
Prostatic binding protein is a complex glycoprotein comprising three components, C1, C2 and C3, organized into two different heterodimers (C1-C3 and C2-C3). The rat ventral prostate genes encoding all three constituent polypeptides are expressed under androgenic control. Analysis of genomic fragments containing the genes and flanking sequences revealed in each case one androgen receptor-binding region upstream of or within the promoter and another in the first intron. The effect of androgens on the expression of these genes may, therefore, be mediated by these direct receptor-DNA interactions. The genomic fragments which contain androgen receptor-binding regions all contain nucleotide sequences reminiscent of glucocorticoid response elements (GRE). Mutations in these sequences in restriction fragments and in synthetic oligonucleotides significantly decreased their affinity for androgen-receptor complexes and their introduction into nonspecific sequences conferred affinity for androgen-receptor complexes. Based on these data, a consensus sequence for putative androgen response elements (ARE) is proposed. However, despite the specific recognition of these sequences by the androgen receptor in vitro, only the C3(1) intronic fragment could confer significant androgen responsiveness on a heterologous promoter. While this could be due to the fact that the GRE-like sequences present in the other fragments are not strong AREs, alternative hypotheses are being investigated currently. Not least of these is that the similar localization of the binding sites in each gene might underlie a more complex androgen regulation mechanism.
Assuntos
Proteína de Ligação a Androgênios/genética , Androgênios/metabolismo , Receptores Androgênicos/metabolismo , Proteína de Ligação a Androgênios/metabolismo , Androgênios/genética , Animais , Sequência de Bases , Regulação da Expressão Gênica , Genes , Glicoproteínas/genética , Glicoproteínas/metabolismo , Dados de Sequência Molecular , Mutação , Prostateína , Ratos , Receptores Androgênicos/genética , Mapeamento por Restrição , Secretoglobinas , UteroglobinaRESUMO
The expression of the three genes encoding the components C1, C2 and C3 of prostatic binding protein (PBP) is under androgen control and restricted to the rat ventral prostate. The SstI-PvuII fragment of the first intron of the C3(1) gene displays two binding sites for ubiquitous transcription factors and one for a tissue-specific factor in a 80-bp region upstream of its androgen response element (ARE). The octamer transcription factor 1 (OTF-1) binds to the most distal element (site 1) while a member of the nuclear factor I (NF-I) family recognizes site 2. A third unidentified prostate-specific factor, which also occurs in castrated rats, interacts with the proximal element (site 3). In T-47D cells, both the OTF-1 and the NF-I-like factor can modulate the androgen response of the promoter in a reporter gene construct containing the C3(1) intronic fragment.
Assuntos
Proteína de Ligação a Androgênios/genética , Androgênios/fisiologia , Proteínas Estimuladoras de Ligação a CCAAT , Proteínas de Ligação a DNA/metabolismo , Genes Reporter/genética , Íntrons/genética , Fatores de Transcrição/metabolismo , Proteína de Ligação a Androgênios/análise , Proteína de Ligação a Androgênios/metabolismo , Animais , Sequência de Bases , Células Cultivadas/química , Células Cultivadas/metabolismo , DNA/análise , DNA/genética , Proteínas de Ligação a DNA/genética , Fator C1 de Célula Hospedeira , Pulmão/química , Pulmão/metabolismo , Masculino , Dados de Sequência Molecular , Fatores de Transcrição NFI , Fator 1 de Transcrição de Octâmero , Regiões Promotoras Genéticas/genética , Próstata/química , Próstata/metabolismo , Prostateína , Ligação Proteica , Ratos , Ratos Wistar , Secretoglobinas , Glândulas Seminais/química , Glândulas Seminais/metabolismo , Testículo/química , Testículo/metabolismo , Fatores de Transcrição/genética , Transcrição Gênica/genética , Transcrição Gênica/fisiologia , Uteroglobina , Proteína 1 de Ligação a Y-BoxRESUMO
Comparable fragments of the androgen receptor (AR) (amino acids 540-607) and of the glucocorticoid receptor (GR) (amino acids 412-515) were expressed in E. coli as fusion proteins with protein A. Both fusion proteins, denoted ARF1 and GRF1, contain the DNA-binding domain and some flanking amino acids. In vitro binding assays have shown that both fusion proteins interact with androgen/glucocorticoid response elements (ARE/GREs) in an intron fragment of the C3(1) gene of the androgen-regulated rat prostatic binding protein and in the typically glucocorticoid-responsive long terminal repeat (LTR) promoter of mouse mammary tumour virus. Present results indicate that the interaction of both ARF1 and GRF1 with the C3(1) as well as the LTR fragments is enhanced in the presence of nuclear extract. The factor that gives rise to this enhancement appears to be ubiquitous and sensitive to trypsin and temperature treatment. In the C3(1) fragment, the enhancing effect requires the presence of an intact functional ARE/GRE (Core II) as well as a region spanning the ARE/GRE half-site Core I.
Assuntos
Androgênios/farmacologia , Núcleo Celular/fisiologia , DNA/metabolismo , Glucocorticoides/farmacologia , Receptores Androgênicos/metabolismo , Receptores de Glucocorticoides/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Íntrons , Masculino , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Regiões Promotoras Genéticas , Próstata/química , Ratos , Ratos Wistar , Receptores Androgênicos/química , Receptores Androgênicos/genética , Receptores de Glucocorticoides/química , Receptores de Glucocorticoides/genética , Sequências Repetitivas de Ácido Nucleico , Relação Estrutura-Atividade , Temperatura , Tripsina/farmacologiaRESUMO
Testicular tumors in infants are rare and most of them are malignant. Hemangioma of the testis is an extremely rare tumor in infants. We report on an intratesticular capillary hemangioma that appeared in a 10 month-old child subsequently undergoing orchidectomy and on the patient's evolution at 4 years. The histological characteristics showed proliferation of small blood vessels, lined with endothelial cells. Mitoses and atypias were not found. Permitting differential diagnosis with other vascular tumors: Masson's vegetant intravascular hemangio-endothelioma and angiosarcoma.
Assuntos
Hemangioma Capilar/patologia , Neoplasias Testiculares/patologia , Testículo/patologia , Capilares/ultraestrutura , Hemangioma Capilar/cirurgia , Hemangioma Capilar/ultraestrutura , Humanos , Lactente , Masculino , Neoplasias Testiculares/cirurgia , Neoplasias Testiculares/ultraestrutura , Testículo/cirurgia , Testículo/ultraestruturaAssuntos
Proteína de Ligação a Androgênios/genética , Androgênios/genética , Proteína de Ligação a Androgênios/metabolismo , Androgênios/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , DNA , Dados de Sequência Molecular , Prostateína , Ratos , Receptores Androgênicos/metabolismo , Secretoglobinas , UteroglobinaAssuntos
Núcleo Celular/metabolismo , DNA/metabolismo , Receptores Androgênicos/metabolismo , Receptores de Glucocorticoides/metabolismo , Animais , Humanos , Mutação , Fragmentos de Peptídeos/metabolismo , Receptores Androgênicos/química , Receptores Androgênicos/genética , Receptores de Glucocorticoides/química , Receptores de Glucocorticoides/genéticaRESUMO
La piscicultura es una actividad importante para satisfacer la demanda alimenticia de una población en crecimiento y en su práctica es necesario desarrollar tecnologías responsables con el ambiente. El policultivo de tilapia-bocachico es relevante por la importancia de estas especies a nivel nacional; tilapia por sus características zootécnicas, y bocachico por su hábito alimentario detritívoro-iliófago, recientemente visto como un pez ecológico. El cultivo basado en perifiton permitiría un sistema altamente eficiente y más limpio, pues aporta alimento natural a los peces y mejora la calidad del agua. Para evaluar el efecto del perifiton sobre el desempeño del policultivo tilapia-bocachico, se ejecutó un experimento durante ocho meses en 18 estanques de 90 m², con 2,6 tilapias/m² y 0,7 bocachicos/m², sin fertilización; como sustrato para perifiton se instalaron de forma vertical tubos plásticos (3,3 tubos/m²), que se constituyeron en una variable por evaluar. Adicionalmente, tres estrategias alimentarias fueron evaluadas: 0 (sin alimento), alimentación con ración del 20% de proteína bruta (PB) y alimentación con ración del 25% de PB. Se utilizó un diseño factorial 2x3 en bloques completos al azar con tres repeticiones por tratamiento y se analizaron datos productivos mediante ANOVA con un modelo lineal general (P < 0,05). No fueron encontrados efectos del sustrato sobre los parámetros estudiados. Hubo efectos simples de la estrategia alimentaria sobre el desempeño de los peces. Por lo pronto, puede ser concluido que la producción basada en perifiton no genera problemas en el sistema ni en los peces y que se requiere más investigación para consolidar la tecnología.
Fishculture is very important to satisfy human food demand. However, the ignorance in more efficient and environment-friendly systems is a big problem for fish farming activities. Tilapia-Bocachico polyculture is of great interest; tilapia, for the excellent productive characteristics, and bocachico for being a bottom-feeder specie, beside of the great socio-cultural value in this region. An alternative approach is the adoption of periphyton-based pond systems; periphyton can: be a food source for fish and improve water quality, thus becoming into a highly efficient and cleaner system. With the aim to evaluate the periphyton effects on performance of Tilapia-Bocachico polyculture, a trial was carried out for 8 months. Eighteen 90 m² earthen ponds were used under polyculture conditions at low stocking density (2.6 tilapia/m², 0.7 bocachico/m²). These ponds were not fertilized during the experimental period. Plastic tubes of 1.2 m and 6 cm in diameter were used vertically as periphyton substrate at 3.3 tubes/m² rate. Three alimentary strategies were used: 0 (without food), fish feed (20% protein) and fish feed (25% protein); thereby constituting a factorial experiment 2x3 in a randomized complete block with three replications per treatment. They were analyzed using ANOVA with a general linear model (GLM) (p < 0.05). There were no effects of substrates on the study variables, but simple effects of alimentary strategies on the productive parameters. Other parameters are being analyzed and it is still premature to conclude.
RESUMO
Fatty acid composition of subcutaneous cellular tissue in 22 children at different ages was determined. The obtained results served as reference values and as indicators of fatty acid composition change with time. Children below and above 45 days of age were compared. Palmitoleic acid showed the greatest variation in composition change with age in the two first months of life. The results are discussed.
Assuntos
Tecido Adiposo/análise , Ácidos Graxos/análise , Fatores Etários , Criança , Fenômenos Fisiológicos da Nutrição Infantil , Pré-Escolar , Humanos , Lactente , Recém-NascidoRESUMO
We demonstrate that the 204 bp intronic gene fragment of C3(1), which has a specific in vitro affinity for the androgen receptor, is able to confer androgen responsiveness to a heterologous promoter. This characteristic is completely destroyed by a single G----T substitution, affecting a 5'-TGTTCT-3' element that closely resembles the consensus sequence of the glucocorticoid and progesterone response elements (GRE/PRE). In fact we could show that this androgen response element (ARE) also acts as a similarly weak GRE or PRE in T-47D cells.
Assuntos
Proteína de Ligação a Androgênios/genética , Genes , Íntrons , Receptores Androgênicos/metabolismo , Animais , Sequência de Bases , Cloranfenicol O-Acetiltransferase/genética , Biblioteca Gênica , Masculino , Plasmídeos , Regiões Promotoras Genéticas , Prostateína , Ligação Proteica , Ratos , Mapeamento por Restrição , Secretoglobinas , Transfecção , UteroglobinaRESUMO
OBJECTIVE: To examine the reactivity pattern and T cell receptor (TCR) characteristics of mycobacterial heat-shock protein 65 (hsp65)-reactive T cells generated from paired synovial fluid (SF) and peripheral blood (PB) samples obtained from rheumatoid arthritis (RA) patients and from healthy subjects. METHODS: The reactivity pattern of hsp65-reactive T cell clones generated under limiting-dilution conditions was analyzed in 3H-thymidine incorporation assays. The TCR variable regions of these hsp65-reactive T cells were characterized by polymerase chain reaction with TCR AV- and BV-specific primers and by DNA sequence analysis of the third complementarity-determining region (CDR3). RESULTS: The hsp65-reactive T cells derived both from RA patients and controls preferentially recognized the 1-170 and 303-540 regions of hsp65 and did not cross-react with human hsp60. The hsp65-reactive T cell clones derived from RA patients displayed a restricted TCR AV and BV gene usage, which can be attributed to the limited clonal origin(s) of the independent T cell clones, as evidenced by CDR3 sequence analysis. These clonally expanded T cells were found in both PB and SF and in different inflamed joints of RA patients. CONCLUSION: Our study suggests that there is in vivo clonal activation and expansion of mycobacterial hsp65-reactive T cells in patients with RA.
Assuntos
Artrite Reumatoide/patologia , Antígenos de Bactérias/análise , Artrite Reumatoide/sangue , Proteínas de Bactérias/análise , Chaperonina 60 , Chaperoninas/análise , Chaperoninas/química , Células Clonais/química , Células Clonais/fisiologia , Humanos , Articulação do Joelho/citologia , Fragmentos de Peptídeos/fisiologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Análise de Sequência de DNA , Líquido Sinovial/citologia , Linfócitos T/química , Linfócitos T/citologiaRESUMO
The purpose of this study was to evaluate the killing effect that treatment in gravity or high-vacuum steam autoclaves had on endospores present on strips or applied to dental needles within 10 types of small sharps containers. Spore strips containing Bacillus stearothermophilus endospores were used, while needles were soiled with an equal number of spores or with spores mixed with blood. Needles were tested capped and uncapped. Strips and needles were autoclaved in empty and 3/4 filled containers placed in several orientations (vents open or closed, upward or on-side). If sterilization was not accomplished within the initial period, additional exposure time was added up to a one hour maximum. Major findings for gravity autoclaves were: 1) strips and needles present in empty containers could, in most cases, be sterilized within 15 minutes when the vents were left open and the containers were placed on-side; 2) spore- and spore plus blood-soiled needles were more difficult to sterilize then were commercial spore strips; 3) capping of needles and the presence of blood did not decrease sterilization efficiency; 4) sterilization in the presence of fill material required additional exposure times and 5) larger containers (greater than 500 ml volume) were more difficult to sterilize than were the smaller (250 ml) ones. All type of open-vented, filled containers could be sterilized within 10 minutes in a high-vacuum autoclave.
Assuntos
Instrumentos Odontológicos , Esterilização/métodos , Geobacillus stearothermophilus , Agulhas , Esporos Bacterianos , VaporRESUMO
Transcription of the genes that code for the different components of Prostatic Binding Protein is dependent on androgens. DNA-cellulose competition assays have demonstrated the presence of regions with affinity for the androgen receptor located immediately upstream and in the first intron of these genes. DNase I footprinting in vitro now reveals the exact binding site of a receptor dimer in the first intron of the C3(1), C3(2) and C1 genes. Gene transfer experiments resulted in the description of functional androgen response elements in the C3 genes. The C1 intronic binding site, however, has a much weaker affinity for the DNA-binding domain of the androgen receptor, a characteristic that might explain its inability to function as an androgen response element in chimeric gene constructs.