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INTRODUCTION: Disparity in the allocation of medical services and resources based on race is present within the health care industry today including the prescription of postoperative analgesics. The purpose of this study was to evaluate the presence of race-based disparity in the prescription of post discharge opioids following lower extremity bypass (LEB) surgery for chronic limb-threatening ischemia (CLTI). METHODS: Retrospective analysis was conducted on adult CLTI patients who underwent LEB from 2000 to 2023 in the TrinetX database. Patients were stratified into two groups based on race: White (Group I) and black or African American (Group II). Primary outcomes were defined as oral opioid prescriptions at 7 days and 30 days post discharge, and mortality at 1 year postoperatively. Secondary outcomes included length of stay (LOS) and 30-day postoperative outcomes including myocardial infarction (MI), pulmonary embolism (PE), cerebral vascular accident (CVA), deep vein thrombosis (DVT), acute kidney injury (AKI), major amputation, minor amputation, Major Adverse Cardiac Events (MACE), and Major Adverse Limb Events (MALE). Stratified analysis was conducted based on disease stage (rest pain vs lower extremity ulcer vs gangrene). Univariate analysis was performed via two-sample t-test and Chi-squared test. Logistic regression was performed to estimate the association of Black or AA (vs. White) race while controlling for pertinent preoperative potential confounders. RESULTS: 3,345 patients met inclusion criteria. Group I included 2,661 White patients and Group II included 684 Black or African American patients. Group II patients were more likely to be younger, female, present with gangrene, and have a history of hypertension, diabetes, chronic kidney disease, or diabetic neuropathy. At both seven- and thirty-days post discharge, the Black or AA cohort had significantly lower rates of opioid prescriptions (33.2% vs 42.5% and 35.8% vs 47.2%, respectively) (all p<0.05). Stratification by indication showed that opioid prescription disparity persisted despite black or AA patients presenting at worse stages of disease both at seven and thirty days post discharge (7-days: Rest pain 43.4% vs. 33.7%, p=0.013, Ulcer 41.4% vs 31.7%, p=0.027, Gangrene, 42.7% vs 33.6%, p=0.006 & 30-days: Rest pain 47.8% vs. 37.1%, p=0.007, Ulcer 45.4% vs 33.5%, p=0.007, Gangrene, 48.2% vs 36.1%, p<0.001). Adjusted analysis confirmed that Black or African American race was associated with lower rates of seven (AOR 0.607, p=0.001) and thirty-day (AOR 0.56, p=0.001) post discharge opioid prescriptions. CONCLUSION: Black or African American patients were less likely to receive post discharge opioid prescriptions compared with their white counterparts at seven- and 30-days following LEB for CLTI.
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Obesity is a major public health concern, and complementary research strategies have been directed toward the identification of the underlying causative gene mutations that affect the normal pathways and networks that regulate energy balance. Here, we describe an autosomal-recessive morbid-obesity syndrome and identify the disease-causing gene defect. The average body mass index of affected family members was 48.7 (range = 36.7-61.0), and all had features of the metabolic syndrome. Homozygosity mapping localized the disease locus to a region in 3q29; we designated this region the morbid obesity 1 (MO1) locus. Sequence analysis identified a homozygous nonsense mutation in CEP19, the gene encoding the ciliary protein CEP19, in all affected family members. CEP19 is highly conserved in vertebrates and invertebrates, is expressed in multiple tissues, and localizes to the centrosome and primary cilia. Homozygous Cep19-knockout mice were morbidly obese, hyperphagic, glucose intolerant, and insulin resistant. Thus, loss of the ciliary protein CEP19 in humans and mice causes morbid obesity and defines a target for investigating the molecular pathogenesis of this disease and potential treatments for obesity and malnutrition.
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Proteínas de Ciclo Celular/genética , Inativação Gênica , Obesidade Mórbida/genética , Adulto , Sequência de Aminoácidos , Animais , Clonagem Molecular , Consanguinidade , Sequência Conservada , Modelos Animais de Doenças , Feminino , Ordem dos Genes , Marcação de Genes , Estudos de Associação Genética , Ligação Genética , Genótipo , Teste de Tolerância a Glucose , Humanos , Insulina/metabolismo , Resistência à Insulina/genética , Masculino , Camundongos , Camundongos Knockout , Dados de Sequência Molecular , Mutação , Obesidade Mórbida/diagnóstico , Linhagem , Fenótipo , Mapeamento Físico do Cromossomo , Transdução de Sinais , Adulto JovemRESUMO
BACKGROUND: Hordeum chilense, a native South American diploid wild barley, is one of the species of the genus Hordeum with a high potential for cereal breeding purposes, given its high crossability with other members of the Triticeae tribe. Hexaploid tritordeum (×Tritordeum Ascherson et Graebner, 2n=6×=42, AABBH(ch)H(ch)) is the fertile amphiploid obtained after chromosome doubling of hybrids between Hordeum chilense and durum wheat. Approaches used in the improvement of this crop have included crosses with hexaploid wheat to promote D/H(ch) chromosome substitutions. While this approach has been successful as was the case with triticale, it has also complicated the genetic composition of the breeding materials. Until now tritordeum lines were analyzed based on molecular cytogenetic techniques and screening with a small set of DNA markers. However, the recent development of DArT markers in H. chilense offers new possibilities to screen large number of accessions more efficiently. RESULTS: Here, we have applied DArT markers to genotype composition in forty-six accessions of hexaploid tritordeum originating from different stages of tritordeum breeding program and to H. chilense-wheat chromosome addition lines to allow their physical mapping. Diversity analyses were conducted including dendrogram construction, principal component analysis and structure inference. Euploid and substituted tritordeums were clearly discriminated independently of the method used. However, dendrogram and Structure analyses allowed the clearest discrimination among substituted tritordeums. The physically mapped markers allowed identifying these groups as substituted tritordeums carrying the following disomic substitutions (DS): DS1D (1H(ch)), DS2D (2H(ch)), DS5D (5H(ch)), DS6D (6H(ch)) and the double substitution DS2D (2H(ch)), DS5D (5H(ch)). These results were validated using chromosome specific EST and SSR markers and GISH analysis. CONCLUSION: In conclusion, DArT markers have proved to be very useful to detect chromosome substitutions in the tritordeum breeding program and thus they are expected to be equally useful to detect translocations both in the tritordeum breeding program and in the transference of H. chilense genetic material in wheat breeding programs.
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Hordeum/genética , Hibridização Genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Ploidias , Triticum/genética , GenótipoRESUMO
Acute type A aortic dissection (TAAD) with malperfusion syndrome remains a challenging diagnosis and optimal surgical management remains unsettled. We present a case and surgical approach employed for a patient with TAAD and malperfusion syndrome who presented with pulseless bilaterally extremities. Satisfactory outcomes can be achieved with early multidisciplinary collaboration and urgent repair of the aorta and simultaneous efforts to restore blood flow to ischemic tissue.
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Aorta Torácica/cirurgia , Aneurisma da Aorta Torácica/cirurgia , Dissecção Aórtica/cirurgia , Isquemia/etiologia , Extremidade Inferior/irrigação sanguínea , Procedimentos Cirúrgicos Vasculares/métodos , Doença Aguda , Dissecção Aórtica/complicações , Dissecção Aórtica/diagnóstico por imagem , Aorta Torácica/diagnóstico por imagem , Aneurisma da Aorta Torácica/diagnóstico por imagem , Circulação Sanguínea , Tamponamento Cardíaco/etiologia , Angiografia por Tomografia Computadorizada , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Objetivo : Identificar la asociación entre la depresión, ansiedad y estrés con las actitudes ante el confinamiento durante la pandemia por COVID-19 en personal de salud del Hospital Nacional Daniel Alcides Carrión y Hospital Militar Central mediante el empleo de la escala DASS-21. Materiales y métodos : Estudio transversal, observacional y analítico, participaron personal de salud que trabajó en áreas Covid durante la pandemia COVID-19. Se empleó la escala Dass-21 para medir la existencia de estados emocionales de depresión, ansiedad y estrés, y la escala de actitudes adaptada frente al confinamiento para COVID-19. Resultados : Se incluyó 124 participantes, 54% (n=67) de sexo femenino, con una mediana de edad de 34.5 años [Ri]= 16.5 que reportaron una mediana de años de experiencia de 6 (Ri = 17). La mediana del puntaje de la Escala Dass-21 para ansiedad fue de 2 (Ri= 5), la mediana del puntaje de Dass-21 para depresión fue de 1 (Ri=4.5) y la mediana del puntaje de la Escala Dass-21 para estrés fue de 4 (Ri=5). El análisis bivariado entre los valores de los parámetros de la Escala Dass-21 y las demás covariables identificó que el grado de instrucción tenía una relación significativa con la subescala de depresión, ansiedad y estrés. Conclusión : Se encontró asociación entre los parámetros afectivos de la escala sobre actitudes ante el confinamiento y las tres subescalas de DASS-21.
Objective : Identify the association between depression, anxiety and stress with attitudes towards confinement during the COVID-19 pandemic in health staff from the Daniel Alcides Carrion National Hospital and Central Military Hospital by using the DASS-21 scale. Materials and methods : Cross-sectional, observational and analytical study, health staff participated who worked in COVID areas during the Covid-19 pandemic. The Dass-21 scale was used to measure the existence of emotional states of depression, anxiety and stress, and the scale of attitudes adapted to confinement for Covid-19. Results : 124 participants were included, 54% female (n=67), with a median age of 34.5 years [Ri]= 16.5, who reported a median year of experience of 6 (Ri = 17). The median Dass-21 score for anxiety was 2 (Ri = 5), the median Dass-21 score for depression was 1 (Ri = 4.5), and the median Dass-21 scale score for stress was 4 (Ri = 5). The bivariate analysis between the values of the Dass-21 Scale domains and the other covariates identified that the level of education had a highly significant relationship with the depression subscale, anxiety and stress. Conclusion : An association was found between the affective parameters of the scale on attitudes towards confinement and the three subscales of DASS-21.
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GH/STAT5 signaling is desensitized in the liver in adult transgenic mice overexpressing GH; however, these animals present greater body size. To assess whether the STAT5 pathway is active during the growth period in the liver in these animals, and how signaling modulators participate in this process, growing transgenic mice and normal siblings were evaluated. STAT5 does not respond to an acute GH-stimulus, but displays higher basal phosphorylation in the livers of growing GH-overexpressing mice. GH receptor and the positive modulators glucocorticoid receptor and HNF1 display greater abundance in transgenic animals, supporting the activity of STAT5. The negative modulators cytokine-induced suppressor and PTP1B are increased in GH-overexpressing mice. The suppressors SOCS2 and SOCS3 exhibit higher mRNA levels in transgenic mice but lower protein content, indicating that they are being actively degraded. Therefore, STAT5 signaling is increased in the liver in GH-transgenic mice during the growth period, with a balance between positive and negative effectors resulting in accelerated but controlled growth.
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Hormônio do Crescimento/metabolismo , Fígado/crescimento & desenvolvimento , Fígado/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais , Animais , Regulação da Expressão Gênica no Desenvolvimento , Fator de Crescimento Insulin-Like I/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fosforilação , Receptores da Somatotropina/genética , Receptores da Somatotropina/metabolismo , Proteínas Supressoras da Sinalização de Citocina/genética , Proteínas Supressoras da Sinalização de Citocina/metabolismoRESUMO
Abstract Introduction: With the evolution of diagnostic techniques in traumatic brain injury (TBI), the study of neurological injury has made progress based on the concepts of primary and secondary injury, leading to the era of proteomics to understand the complex molecular events involved in the process. Objectives: This narrative review is intended to discuss the state of the art of the most frequently used biomarkers in TBI, their clinical utility, and the implications for therapeutic decision-making protocols. Materials and methods: In order to fulfill the objective of this paper, a literature review was conducted of the most important databases. Results: Several biomarkers have been studied as prognostic factors in patients with TBI. Learning about their sensitivity and specificity in neurological injury, and its post-trauma evolution over time, has been the goal of various papers in the past few years. Conclusion: Breakthroughs in the study of protein degradation make it necessary to broaden the spectrum and knowledge of new diagnostic methods in TBI. Further studies are needed to define the role of biomarkers and to promote protocols integrating specific values.
Resumen Introducción: Con la evolución de las técnicas diagnósticas en el trauma craneoencefálico, el estudio de la lesión neurológica ha progresado sobre los conceptos de lesión primaria y secundaria, para entrar así en la era de la proteómica y, con ella, entender los complejos eventos moleculares existentes en su proceso. Objetivos: En esta revisión narrativa se pretende presentar el estado actual de los biomarcadores que más se usan en lesión cerebral traumática, su utilidad clínica y las implicaciones en protocolos de decisión terapéutica. Materiales y métodos: Para dar respuesta al objetivo de este trabajo, se realizó una revisión de la literatura en las principales bases de datos. Resultados: Se han estudiado varios biomarcadores como factor pronóstico en pacientes con trauma craneoencefálico. Conocer su sensibilidad y especificidad para la lesión neurológica, así como su evolución en el tiempo tras el traumatismo, ha sido el objetivo de diversos trabajos en los últimos años. Conclusión: El avance en el estudio de los productos de degradación de las proteínas hace necesario ampliar el espectro y el conocimiento en el campo de los nuevos métodos diagnósticos en el trauma craneoencefálico. Se requieren más estudios para definir la función de los biomarcadores y proponer protocolos que integren valores específicos.
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Humanos , Biomarcadores , Lesões dos Tecidos Moles , Lesões Encefálicas Traumáticas , Prognóstico , Fatores Biológicos/administração & dosagem , ProteômicaRESUMO
Background: to the best of our knowledge, the effects of lipopolysaccharide (LPS) from Escherichia coli on goblet cells and intestinal mucin secretion of weaned pigs has not been reported, and it is unknown whether these effects could trigger enteritis. Objective: to determine the effect of E. coli LPS on intestinal mucin secretion in weaning piglets. Methods: fifty-two piglets weaned at 21 days of age were fed a basal diet supplemented with four LPS levels (0.0, 0.3, 0.5, and 1.0 μg/mg) during 10 days. Piglets were slaughtered on days 1, 5, 7, and 10 post-weaning and samples of small and large intestine were taken for histochemical staining to determine goblet cell population and type of mucins produced (acidic, sulphated, non-sulphated, or neutral). Results: acidic mucin was reduced on day 5 post-weaning independently of the dietary LPS level supplied to piglets. Recovery of acidic mucins was observed during days 7 and 10 post-weaning. Neutral mucins increased on day 5 and decreased on days 7 and 10 post-weaning. High LPS levels decreased goblet cells population and secretion of all types of mucins. This effect was remarkably high for diet two (D2: 0.5 mg LPS/mg food). Conclusions: early weaning (21 d) and LPS addition to the diet affect mucin secretion and intestinal epithelium integrity by modifying goblet cell populations and their balance between acidic and neutral mucin secretion. These findings explain some abnormalities related with post-weaning diarrhea syndrome and help to explain its pathophysiology.
Antecedentes: actualmente se desconoce el efecto del lipopolisacárido (LPS) de Escherichia coli sobre la cantidad de células caliciformes y la secreción de mucinas en diferentes regiones del intestino en cerdos durante el período pos-destete. Tampoco se ha descrito si cambios en la distribución de las mucinas en el intestino están relacionados con el desarrollo de enteritis. Objetivo: determinar el efecto del LPS de E. coli sobre la secreción de mucinas en el intestino de lechones recién destetados. Métodos: cincuenta y dos lechones destetados a los 21 días fueron alimentados con una dieta basal adicionada con cuatro niveles de LPS (0,0, 0,3, 0,5 y 1,0 μg/mg) durante 10 días. Los cerdos se sacrificaron los días 1, 5, 7 y 10 pos-destete y se tomaron muestras de intestino delgado y colon para realizar coloraciones histoquímicas, que permitieran calcular la cantidad de células caliciformes y el tipo de mucinas ácidas sulfatadas, no sulfatadas o neutras por ellas producidas. Resultados: la producción de mucinas ácidas en las células caliciformes se redujo el día 5 del período pos-destete, con posterior restauración de los parámetros a los días 7 y 10 e independientemente de la dosis de LPS suministrada en la dieta. En contraste, la producción de mucinas neutras aumentó en el día 5 y disminuyó en los días 7 y 10 del período pos-destete. Al comparar las dietas experimentales, se observó que dosis mayores de LPS, disminuyen el número de células caliciformes y la secreción de los diferentes tipos de mucinas. Este efecto fue más marcado con la dieta dos (D2: 0,5 mg de LPS/mg de alimento). Conclusiones: el destete a los 21 días y la adición LPS de E. coli a la dieta generan cambios en la secreción de mucinas, afectan la integridad del epitelio y el balance entre la secreción de mucinas ácidas y neutras por las células caliciformes. Estos hallazgos sugieren explicaciones de algunas alteraciones que se producen en el síndrome de diarrea pos-destete y contribuyen a explicar su fisiopatología.
Antecedentes: actualmente é desconhecido o efeito do lipopolissacarídeo (LPS) de Escherichia coli sobre a quantidade de células caliciformes, sobre a secreção de mucinas em diferentes regiões do intestino em porcos durante o período de pós-desmame e se o padrão de distribuição das mucinas está relacionado com o desenvolvimento de enterite. Objetivo: determinar o efeito da LPS de E. coli sobre a secreção de mucinas no intestino de leitões recém desmamados. Métodos: foi realizado um estudo experimental com 52 leitões desmamados aos 21 dias, que foram alimentados com uma dieta basal adicionada com quatro níveis de LPS (0,0, 0,3, 0,5 y 1,0 μg/mg) durante 10 dias. Os porcos foram sacrificados os dias 1, 5, 7 e 10 pós-desmame para tirar amostras de intestino delgado e grosso, realizar colorações histoquímicas, calcular a quantidade de células caliciformes e o tipo de mucinas ácidas sulfatadas, não sulfatadas e neutras por elas produzidas. Resultados: observou-se que ao número de células caliciformes que expressaram mucinas ácidas nos diferentes tempos do período pós-desmame, apresentaram uma diminuição da secreção de mucinas ácidas no quinto dia, com posterior recuperação dos parâmetros os dias 7 e 10 Independentemente da dose de LPS disso. Ao contrário, as mucinas neutras incrementaram o dia 5 com uma posterior diminuição os dias 7 e 10 pós-desmame. Ao comparar as dietas experimentais, observou-se uma diminuição do número de células caliciformes secretando os diferentes tipos de mucinas a doses maiores de LPS, principalmente com a dieta dois (D2: 0,5 mg LPS/mg comida). Conclusões: o desmame aos 21 dias e a adição de LPS de E. coli na dieta, gera mudanças na secreção de mucinas, afetam a integridade do epitélio e o balanço entre a secreção de mucinas ácidas e neutras pelas células caliciformes; isto modela algumas alterações que se produzem no síndrome de diarreia pós-desmame e contribui a explicar a sua fisiopatologia.
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Early weaning predisposes the pig intestine to structural and functional alterations, due to the increase in E. coli populations. These bacteria use the lipopolysaccharide (LPS) derived from their cell wall as an important pathogenic factor. Little is known about the effects of LPS on the intestinal morphology. Such knowledge could be helpful in understanding the pathogenesis of post-weaning enteritis, which is needed to design therapeutic strategies. Objective: this study aimed to evaluate the effects of the oral intake of LPSon the morphology of intestinal villi and glands of weaned pigs. Methods: the study used 52 pigs weaned at 21 days. The animals were fed a basal diet added with four levels of LPS (0.0, 0.3, 0.5 and 1.0 µg/mg of food) for 10 days. Pigs were sequentially slaughtered on days 1, 5, 7 and 10 after weaning, and samples of small intestine were taken to evaluate morphological parameters by computerized image analysis. The statistical design used was randomized blocks in a 4x4 factorial arrangement. Results: results showed that LPS decreases the height and area of intestinal villi, and increases the width of the villi and the depth and width of the intestinal glands. These effects probably contribute to a decreased intestinal nutrient absorption and increase co-infection with other pathogens, thus leading to the post-weaning diarrhea syndrome. Conclusions: this study stresses the usefulness of computerized morphometric analysis to evaluate the effect of LPS on intestinal morphology, so it may be used in future studies to investigate the pathophysiology of the causative agents of enteritis and to evaluate therapeutic strategies.
El destete precoz de los cerdos predispone al desarrollo de alteraciones estructurales y funcionales en el intestino y a enteritis causadas por la bacteria Escherichia coli; la cual utiliza el LPS de su pared como uno de sus principales factores patogénicos. Debido a que se conoce poco sobre los efectos del LPS sobre los parámetros morfológicos intestinales, y a que ese conocimiento es necesario para comprender la patogenia de las enteritis postdestete y para diseñar estrategias terapéuticas. Objetivo: se realizó este estudio con el objetivo de evaluar el efecto de la administración de LPS de E. coli sobre la morfología de las vellosidades y las glándulas intestinales en cerdos recién destetados. Métodos: El estudio experimental se realizó con 52 cerdos destetados a los 21 días de edad. Los animales fueron alimentados con una dieta basal adicionada con cuatro niveles de LPS (0.0, 0.3, 0.5 y 1.0 µg/mg de alimento) durante 10 días. Los cerdos se sacrificaron escalonadamente los días 1, 5, 7 y 10 posdestete y se tomaron muestras de intestino delgado para determinar algunos parámetros morfológicos mediante análisis computarizado de imágenes. El diseño estadístico empleado fue bloques al azar en un arreglo factorial 4x4. Resultados: como resultados se obtuvo que el LPS disminuye la altura y el área de las vellosidades y aumenta su ancho, así como la profundidad y ancho de las glándulas intestinales. Estos efectos probablemente disminuyen la absorción intestinal de nutrientes, incrementan la co-infección con otros agentes patógenos y la presentación del síndrome de diarrea posdestete. Conclusiones: Este estudio muestra la utilidad del análisis morfométrico computarizado para evaluar el efecto del LPS sobre los parámetros morfológicos intestinales, por lo que podría utilizarse en futuros estudios para investigar la fisiopatología de los agentes causantes de enteritis y para evaluar estrategias terapéuticas.
O desmame precoce dos suínos predispõe o desenvolvimento de alterações estruturais e funcionais no intestino e à enterite causada pela bactéria Escherichia coli, que usa o LPS da parede como um dos principais fatores patogênicos. Devido a que pouco se sabe sobre os efeitos do LPS sobre os parâmetros morfológicos intestinais, e que esse conhecimento é necessário para compreender a patogênese da enterite pós-desmame e projetar estratégias terapéuticas. Objetivo: este estudo foi realizado para avaliar o efeito administração de LPS de E. coli sobre a morfologia das vilosidades e glândulas intestinais em suínos desmamados. Métodos: o estudo experimental foi realizado com 52 leitões desmamados aos 21 dias de idade. Os animais foram alimentados com uma dieta basal suplementada com quatro níveis de LPS (0.0, 0.3, 0.5 e 1.0 µg/mg de alimento) durante 10 dias. Os suínos foram abatidos em escalonadamente aos 1, 5, 7 e 10 dias pós-desmame e foram tomadas amostras do intestino delgado para determinar alguns parâmetros morfológicos através da análise computacional de imagens. O delineamento estatístico utilizado foi em blocos casualizados em um arranjo fatorial 4x4. Resultados: o resultado foi que LPS diminuiu a altura e a área das vilosidades e aumenta sua largura e profundidade e amplitude das glândulas intestinais. Estes efeitos podem diminuir a absorção intestinal de nutrientes, aumento de co-infecção com outros patógenos ea apresentação do pós-desmame síndrome diarréica. Conclusões: este estudo mostra a utilidade da análise morfométrica computadorizada para avaliar o efeito do LPS sobre parâmetros morfológicos intestinais, de modo que poderiam ser utilizados em futuros estudos para pesquisar a fisiopatologia da enterite agentes causadores e avaliar estratégias terapêuticas.
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Congenital erythropoietic porphyria (CEP), an autosomal recessive inborn error, results from the deficient but not absent activity of uroporphyrinogen III synthase (URO-synthase), the fourth enzyme in the heme biosynthetic pathway. The major clinical manifestations include severe anemia, erythrodontia, and disfiguring cutaneous involvement due to the accumulation of phototoxic porphyrin I isomers. Murine models of CEP could facilitate studies of disease pathogenesis and the evaluation of therapeutic endeavors. However, URO-synthase null mice were early embryonic lethals. Therefore, knock-in mice were generated with three missense mutations, C73R, V99A, and V99L, which had in vitro-expressed activities of 0.24%, 5.9%, and 14.8% of expressed wild-type activity, respectively. Homozygous mice for all three mutations were fetal lethals, except for mice homozygous for a spontaneous recombinant allele, V99A(T)/V99A(T), a head-to-tail concatemer of three V99A targeting constructs. Although V99A(T)/V99A(T) and C73R/V99A(T) mice had approximately 2% hepatic URO-synthase activity and normal hepatic microsomal heme and hemoprotein levels, they had 20% and 13% of wild-type activity in erythrocytes, respectively, which indicates that sufficient erythroid URO-synthase was present for fetal development and survival. Both murine genotypes showed marked porphyrin I isomer accumulation in erythrocytes, bone, tissues, and excreta and had fluorescent erythrodontia, hemolytic anemia with reticulocytosis and extramedullary erythropoiesis, and, notably, the characteristic light-induced cutaneous involvement. These mice provide insight into why CEP is an erythroid porphyria, and they should facilitate studies of the disease pathogenesis and therapeutic endeavors for CEP.
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Luz/efeitos adversos , Porfiria Eritropoética/genética , Dermatopatias/etiologia , Uroporfirinogênio III Sintetase/fisiologia , Animais , Heme/metabolismo , Humanos , Camundongos , Camundongos Transgênicos , Microssomos Hepáticos/metabolismo , Dados de Sequência Molecular , Fenótipo , Porfiria Eritropoética/enzimologia , Porfiria Eritropoética/fisiopatologia , Porfirinas/metabolismo , Dermatopatias/fisiopatologia , Uroporfirinogênio III Sintetase/genéticaRESUMO
Priming of CD8(+) T cells requires presentation of short peptides bound to MHC class I molecules of professional APCs. Cross-presentation is a mechanism whereby professional APC present on their own MHC class I molecules peptides derived from degradation of Ags synthesized by other Ag "donor cells." The mechanism of cross-presentation is poorly understood, and the nature of the transferred Ag is unknown. In this report, we demonstrate that the bulk of a cross-presented Ag transferred from donor cells recently infected with vaccinia virus are proteasomal products that are susceptible to peptidases within the donor cell cytosol and not full-length proteins or mature epitopes either free or bound to chaperones.
Assuntos
Apresentação de Antígeno/imunologia , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Antígenos/metabolismo , Cisteína Endopeptidases/metabolismo , Antígenos H-2/metabolismo , Complexos Multienzimáticos/metabolismo , Vaccinia virus/imunologia , Animais , Células Apresentadoras de Antígenos/virologia , Linhagem Celular , Galinhas , Citosol/imunologia , Citosol/metabolismo , Proteínas do Ovo/imunologia , Proteínas do Ovo/metabolismo , Retículo Endoplasmático/imunologia , Retículo Endoplasmático/metabolismo , Antígenos H-2/imunologia , Células HeLa , Humanos , Hibridomas , Epitopos Imunodominantes/imunologia , Epitopos Imunodominantes/metabolismo , Camundongos , Chaperonas Moleculares/metabolismo , Ovalbumina/imunologia , Ovalbumina/metabolismo , Fragmentos de Peptídeos , Complexo de Endopeptidases do Proteassoma , Ligação Proteica/imunologia , Transporte Proteico/imunologiaRESUMO
Juvenile hyaline fibromatosis (JHF) and infantile systemic hyalinosis (ISH) are autosomal recessive syndromes of unknown etiology characterized by multiple, recurring subcutaneous tumors, gingival hypertrophy, joint contractures, osteolysis, and osteoporosis. Both are believed to be allelic disorders; ISH is distinguished from JHF by its more severe phenotype, which includes hyaline deposits in multiple organs, recurrent infections, and death within the first 2 years of life. Using the previously reported chromosome 4q21 JHF disease locus as a guide for candidate-gene identification, we identified and characterized JHF and ISH disease-causing mutations in the capillary morphogenesis factor-2 gene (CMG2). Although CMG2 encodes a protein upregulated in endothelial cells during capillary formation and was recently shown to function as an anthrax-toxin receptor, its physiologic role is unclear. Two ISH family-specific truncating mutations, E220X and the 1-bp insertion P357insC that results in translation of an out-of-frame stop codon, were generated by site-directed mutagenesis and were shown to delete the CMG-2 transmembrane and/or cytosolic domains, respectively. An ISH compound mutation, I189T, is predicted to create a novel and destabilizing internal cavity within the protein. The JHF family-specific homoallelic missense mutation G105D destabilizes a von Willebrand factor A extracellular domain alpha-helix, whereas the other mutation, L329R, occurs within the transmembrane domain of the protein. Finally, and possibly providing insight into the pathophysiology of these diseases, analysis of fibroblasts derived from patients with JHF or ISH suggests that CMG2 mutations abrogate normal cell interactions with the extracellular matrix.
Assuntos
Fibroma/genética , Glomerulosclerose Segmentar e Focal/genética , Proteínas de Membrana/genética , Sequência de Aminoácidos , Sequência de Bases , Criança , Mapeamento Cromossômico , Éxons , Feminino , Genes Recessivos , Marcadores Genéticos , Humanos , Lactente , Masculino , Modelos Moleculares , Mutação de Sentido Incorreto , Linhagem , Conformação Proteica , Estrutura Secundária de Proteína , Receptores de Peptídeos , SíndromeRESUMO
Se utiliza una técnica de coloración con plata para visualizar las Regiones Organizadoras Nucleolares (AgNORs). La evaluación cuantitativa y cualitativa de estas regiones representa en la actualidad un marcador de actividad proliferativa en células tumorales. El presente estudio tuvo como objetivo caracterizar las AgNORs en algunos tipos de tumores cutáneos caninos, para lo cual se evaluaron 28 mastocitomas, 18 carcinomas espinocelulares y siete epiteliomas basocelulares, procedentes del archivo de Patología Animal y del Consultorio Veterinario de la Universidad de Antioquia, y de otros consultorios de la ciudad de Medellín, Colombia. Las muestras bloqueadas en parafina se cortaron a cuatro micras, y se colorearon con Hematoxilina-Eosina y Giemsa paradiagnosticar y clasificar los mastocitomas. Se utilizó otra serie de cortes coloreados con plata para evaluar morfométricamente las AgNORs utilizando un Sistema Automático Analizador de Imágenes (SAAI). Se evaluaron los parámetros: área nuclear, área AgNORs/célula, número de AgNORs/ célula, y distribución AgNORs en la célula. Para determinar el número de células a evaluar se utilizó el estudio de variación de la inestabilidad de los valores medios, con relación al tamaño de la muestra;se obtuvo un mínimo representativo de 20 células por caso para los mastocitomas y los carcinomas espinocelulares, y de 30 células por caso, para los tumores basocelulares. Los datos fueron analizadosestadísticamente mediante el análisis de varianza (ANOVA) y se compararon las medias por el test de Fischer (F) empleando un nivel de significancia de p<0.05. La coloración AgNORs fue factible en lostumores evaluados para lo cual se necesitaron de 30 minutos en el período de incubación. El análisis estadístico mostró que los mastocitomas grado II poseen un área nuclear menor estadísticamentesignificativa (p<0.05). Existe además, diferencia estadística significativa (p<0.05) en el número y en el área de las AgNORs entre los tres grados hi...