RESUMO
Despite the high incidence of major depressive disorder (MDD) in females, the detailed neurobiology mechanism remains not fully understood. Increasing evidence showed that MDD was associated with hippocampal volumetric abnormality with different subfields demonstrating various alteration features. However, the linkage between hippocampal atrophy with its biochemical information remains unclear. In this study, we aim to investigate the relationship between bilateral hippocampal subfields volumetric and metabolic information in female MDD patients, using a combined T1weighted magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (H-MRS) technology. There are 15 female MDD patients and 12 matched healthy controls involved in the study. We found a significant decrease in left fimbria volume in MDD group, which was negatively correlated with left hippocampal choline level. In addition, the left hippocampal creatine concentration in patients was negatively correlated with left fimbria volume. Moreover, the NAA level in left hippocampus was negatively correlated with MDD clinical symptomology including anxiety and depression scores. Our findings suggest that the altered coupling between hippocampal structural and metabolic features might contribute to the etiology of female MDD patients.
Assuntos
Transtorno Depressivo Maior , Atrofia , Creatina , Feminino , Hipocampo , Humanos , Imageamento por Ressonância Magnética , Espectroscopia de Prótons por Ressonância MagnéticaRESUMO
Mycoplasma gallisepticum (MG) is a member of the most important avian mycoplasmas, causing chronic respiratory disease in chickens and leading to important economic losses in the poultry industry. Recombinant technology represents a strategic approach used to achieve highly reliable and specific diagnostic tests in veterinary diseases control: in particular this aspect is crucial for confirming mycoplasma infection and for maintaining mycoplasma-free breeder flocks. In this study, we identified a component of the pyruvate dehydrogenase dihydrolipoamide acetyltransferase (i.e., E2) protein by 2-dimensional electrophoresis (2-DE), characterized it in immunoblotting assays, and analyzed its recombinant (r-E2) in a rec-ELISA test. For full-length protein expression in Escherichia coli (EC) a point mutation was introduced. A rabbit antiserum produced against r-E2 was tested in a Western Blot using different samples of Mycoplasma species. The results showed the applicability of site-directed mutagenesis, with a good yield of the r-E2 after purification. Also, anti-E2 serum reacted with all the tested MG strains showing no cross reaction with other mycoplasmas. The developed E2 ELISA test was capable of detecting MG antibodies in the sera examined. Those results demonstrate the antigenic stability of the E2 protein which could represent a recombinant antigen with potential diagnostic applications.
Assuntos
Proteínas de Bactérias/genética , Galinhas/microbiologia , Mycoplasma gallisepticum/genética , Animais , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática/veterinária , Escherichia coli/genética , Expressão Gênica , Vetores Genéticos/genética , Mycoplasma gallisepticum/imunologia , Mycoplasma gallisepticum/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismoRESUMO
OBJECTIVE: Not exclusively breastfed children with cow's milk allergy (CMA) require a formula or other alternative food, but past and present guidelines differ concerning the best choice. Our aim was to investigate the clinical tolerability, palatability and nutritional adequacy of donkey's milk (DM) in children with proven CMA. It was important to identify a CM replacement for these children, highly problematic from the feeding standpoint, in spite of their age. STUDY DESIGN: A prospective study was conducted on 92 children with CMA, diagnosed through a CM elimination diet, followed by double-blind, placebo-controlled food challenge (DBPCFC) unless contraindicated. Maternal milk was unavailable and current CM substitutes could not be used. Moreover, 89 percent were affected by multiple FA, and subjected to very restricted diets. Within 3 months after the last CM challenge, DBPCFC for DM was performed. CM or DM skin prick test and sIgE determination preceded the CM or DM challenge, respectively. Native electrophoresis and immunoblotting were used to identify CM and DM cross-reactive proteins. Z-scores of weight and length/stature for age were calculated at DM food challenge (T0) and during DM assumption. RESULTS: 83 children (90.2 percent) liked and tolerated DM, at challenge and during follow-up, with increased Z-score for weight and length/stature and improved nutritional parameters. Bovine beta-lactoglobulin was identified as the cross-reacting protein among the DM allergic patients. CONCLUSIONS: DM was found to be a valid alternative foodstuff, in terms of clinical tolerability, palatability and nutritional adequacy, in subjects with CMA who were highly problematic from the feeding standpoint.
Assuntos
Fenômenos Fisiológicos da Nutrição do Lactente/imunologia , Lactoglobulinas/imunologia , Hipersensibilidade a Leite/prevenção & controle , Leite , Animais , Peso ao Nascer , Estatura , Bovinos , Criança , Pré-Escolar , Método Duplo-Cego , Equidae , Feminino , Humanos , Immunoblotting , Imunoglobulina E/sangue , Lactente , Lactoglobulinas/sangue , Masculino , Hipersensibilidade a Leite/imunologia , Leite Humano , Placebos , Estudos Prospectivos , Testes CutâneosRESUMO
Poly(ADP-ribose) polymerase-2 (Parp-2) belongs to a family of enzymes that catalyse poly(ADP-ribosyl)ation of proteins. Parp-2 deficiency in mice (Parp-2(-/-)) results in reduced thymic cellularity associated with increased apoptosis in thymocytes, defining Parp-2 as an important mediator of T-cell survival during thymopoiesis. To determine whether there is a link between Parp-2 and the p53 DNA-damage-dependent apoptotic response, we have generated Parp-2/p53-double-null mutant mice. We found that p53(-/-) backgrounds completely restored the survival and development of Parp-2(-/-) thymocytes. However, Parp-2-deficient thymocytes accumulated high levels of DNA double-strand breaks (DSB), independently of the p53 status, in line with a function of Parp-2 as a caretaker promoting genomic stability during thymocytes development. Although Parp-2(-/-) mice do not have spontaneous tumours, Parp-2 deficiency accelerated spontaneous tumour development in p53-null mice, mainly T-cell lymphomas. These data suggest a synergistic interaction between Parp-2 and p53 in tumour suppression through the role of Parp-2 in DNA-damage response and genome integrity surveillance, and point to the potential importance of examining human tumours for the status of both genes.
Assuntos
Linfoma de Células T/metabolismo , Linfoma de Células T/patologia , Poli(ADP-Ribose) Polimerases/deficiência , Proteína Supressora de Tumor p53/deficiência , Animais , Quebras de DNA de Cadeia Dupla , Feminino , Linfoma de Células T/enzimologia , Linfoma de Células T/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Poli(ADP-Ribose) Polimerases/metabolismo , Timo/citologia , Timo/metabolismo , Fatores de Tempo , Proteína Supressora de Tumor p53/metabolismoRESUMO
Initially the resistance to digestion of two cow's milk allergens, beta-casein, and beta-lactoglobulin (beta-Lg), was compared using a "high-protease assay" and a "low-protease assay" in a single laboratory. The low-protease assay represents an alternative standardised protocol mimicking conditions found in the gastrointestinal tract. For the high-protease assay, both proteins were incubated with either pepsin or pancreatin and digestion monitored by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse phase-high performance liquid chromatography. The low-protease assay involved gastroduodenal digestion in the presence or absence of phosphatidylcholine (PC). Both beta-casein and beta-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of beta-casein digestion in the low-protease assay were slower, beta-Lg being pepsin resistant. During duodenal digestion, beta-Lg was gradually degraded and addition of PC slowed digestion. Subsequently, the reproducibility of the low-protease assay was assessed in 12 independent laboratories by visual assessment of the gels and densitometric analysis: the inter- and intra-laboratory variability was affected by sampling and electrophoresis method employed. The low-protease assay was shown to be reproducible. Future studies will extend these findings using a broader panel of proteins.