ABSTRACT
The proline-rich inhibitor of 31 kDa (PI31) is highly conserved through metazoan evolution, and its activity in the proteasome inhibition is well-established although the precise mechanism of inhibition is unclear. The coding DNA sequence of Schistosoma mansoni PI31 (SmPI31) was cloned, and the recombinant protein was expressed in bacterial system. The correct amino acid sequence was confirmed by mass spectrometry and circular dichroism suggests that SmPI31 contains both α-helix and non-structured regions. Inhibition assays, using the Suc-Leu-Leu-Val-Tyr-4-MCA substrate for proteasome degradation, showed that the S. mansoni proteasome may be regulated by the inhibitory activity of SmPI31. A gene expression assay using qRT-PCR at various stages during the S. mansoni life cycle has shown that SmPI31 transcripts are expressed in all studied stages, suggesting that PI31 plays an important role during the developmental processes of the parasite. In this study first evidence is presented that PI31 has a conserved structure and plays a role as proteasome inhibitor in adult worms and it is expressed through life cycle.
Subject(s)
Cysteine Proteinase Inhibitors/biosynthesis , Gene Expression Profiling , Proteasome Inhibitors , Protozoan Proteins/biosynthesis , Schistosoma mansoni/enzymology , Animals , Circular Dichroism , Cloning, Molecular , Cysteine Proteinase Inhibitors/chemistry , Cysteine Proteinase Inhibitors/genetics , Gene Expression , Mice , Mice, Inbred BALB C , Protein Conformation , Protozoan Proteins/chemistry , Protozoan Proteins/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Schistosoma mansoni/geneticsABSTRACT
The interaction between the reproductive axis and energy balance suggests that leptin acts as a possible mediator. This hormone acts in the regulation of metabolism, feeding behaviour and reproduction. Animals homozygous for the gene 'ob' (ob/ob) are obese and infertile, and these effects are reversed after systemic administration of leptin. Thus, the present study aimed to determine: (i) whether cells that express leptin also express oestrogen receptors of type-alpha (ER-alpha) or -beta (ER-beta) in the medial preoptic area (MPOA) and in the arcuate (ARC), dorsomedial (DMH) and ventromedial hypothalamic nucleus and (ii) whether there is change in the gene and protein expression of leptin in these brain areas in ovariectomised (OVX) animals when oestrogen-primed. Wistar female rats with normal oestrous cycles or ovariectomised oestrogen-primed or vehicle (oil)-primed were utilised. To determine whether there was a co-expression, immunofluorescence was utilised for double staining. Confocal microscopy was used to confirm the co-expression. The technique of real-time polymerase chain reaction and western blotting were employed to analyse gene and protein expression, respectively. The results obtained showed co-expression of leptin and ER-alpha in the MPOA and in the DMH, as well as leptin and ER-beta in the MPOA, DMH and ARC. However, we did not detect leptin in the MPOA, ARC and DMH using western blotting and there was no statistical difference in leptin gene expression in the MPOA, DMH, ARC, pituitary or adipose tissue between OVX rats treated with oestrogen or vehicle. In conclusion, the results obtained in the present study confirm that the brain is also a source of leptin and reveal co-expression of oestrogen receptors and leptin in the same cells from areas related to reproductive function and feeding behaviour. Although these data corroborate the previous evidence obtained concerning the interaction between the action of brain leptin and reproductive function, the physiological relevance of this interaction remains uncertain and additional studies are necessary to elucidate the exact role of central leptin.
Subject(s)
Hypothalamus/metabolism , Leptin/genetics , Preoptic Area/metabolism , Receptors, Estrogen/genetics , Animals , Estrogens/pharmacology , Female , Gene Expression/drug effects , Hypothalamus/drug effects , Leptin/metabolism , Preoptic Area/drug effects , RNA, Messenger/metabolism , Rats , Rats, Wistar , Receptors, Estrogen/metabolism , Reproduction/genetics , Tissue Distribution/drug effectsABSTRACT
Apresenta-se caso de lesao do nervo mediano em luxacao posterior do cotovelo, que, devido a sua gravidade, deve ser pensada, quando se lida com crianca que sofreu luxacao do cotovelo, principalmente se houver fratura do epicondilo medial associada. E feita revisao do mecanismo, sendo apresentados sinais clinicos e radiologicos e tecnica cirurgica modificada
