Association of decreased mitochondrial DNA content with ovarian cancer progression

Mitochondrial DNA (mtDNA) content in ovarian carcinomas was assessed by quantitative PCR. Results show that mtDNA content in tumour cell was significantly higher than that in normal ovary. Change in mtDNA content was not related with patients' age or tumour stages. However, the average mtDNA copy number in pathological low-grade tumours was over two-fold higher than that in high-grade carcinomas (P=0.012). Moreover, type I carcinomas also had a significantly higher mtDNA copy number than in type II carcinomas (P=0.019). Change in mtDNA content might be an important genetic event in the progression of ovarian carcinomas

The spin label amino acid TOAC and its uses in studies of peptides: chemical, physicochemical, spectroscopic, and conformational aspects

We review work on the paramagnetic amino acid 2,2,6,6-tetramethyl-N-oxyl-4-amino-4-carboxylic acid, TOAC, and its applications in studies of peptides and peptide synthesis. TOAC was the first spin label probe incorporated in peptides by means of a peptide bond. In view of the rigid character of this cyclic molecule and its attachment to the peptide backbone via a peptide bond, TOAC incorporation has been very useful to analyze backbone dynamics and peptide secondary structure. Many of these studies were performed making use of EPR spectroscopy, but other physical techniques, such as X-ray crystallography, CD, fluorescence, NMR, and FT-IR, have been employed. The use of double-labeled synthetic peptides has allowed the investigation of their secondary structure. A large number of studies have focused on the interaction of peptides, both synthetic and biologically active, with membranes. In the latter case, work has been reported on ligands and fragments of GPCR, host defense peptides, phospholamban, and β-amyloid. EPR studies of macroscopically aligned samples have provided information on the orientation of peptides in membranes. More recent studies have focused on peptide–protein and peptide–nucleic acid interactions. Moreover, TOAC has been shown to be a valuable probe for paramagnetic relaxation enhancement NMR studies of the interaction of labeled peptides with proteins. The growth of the number of TOAC-related publications suggests that this unnatural amino acid will find increasing applications in the future

In vitro nuclear interactome of the HIV-1 Tat protein

<p>Abstract</p> <p>Background</p> <p>One facet of the complexity underlying the biology of HIV-1 resides not only in its limited number of viral proteins, but in the extensive repertoire of cellular proteins they interact with and their higher-order assembly. HIV-1 encodes the regulatory protein Tat (86–101aa), which is essential for HIV-1 replication and primarily orchestrates HIV-1 provirus transcriptional regulation. Previous studies have demonstrated that Tat function is highly dependent on specific interactions with a range of cellular proteins. However they can only partially account for the intricate molecular mechanisms underlying the dynamics of proviral gene expression. To obtain a comprehensive nuclear interaction map of Tat in T-cells, we have designed a proteomic strategy based on affinity chromatography coupled with mass spectrometry.</p> <p>Results</p> <p>Our approach resulted in the identification of a total of 183 candidates as Tat nuclear partners, 90% of which have not been previously characterised. Subsequently we applied <it>in silico </it>analysis, to validate and characterise our dataset which revealed that the Tat nuclear interactome exhibits unique signature(s). First, motif composition analysis highlighted that our dataset is enriched for domains mediating protein, RNA and DNA interactions, and helicase and ATPase activities. Secondly, functional classification and network reconstruction clearly depicted Tat as a polyvalent protein adaptor and positioned Tat at the nexus of a densely interconnected interaction network involved in a range of biological processes which included gene expression regulation, RNA biogenesis, chromatin structure, chromosome organisation, DNA replication and nuclear architecture.</p> <p>Conclusion</p> <p>We have completed the <it>in vitro </it>Tat nuclear interactome and have highlighted its modular network properties and particularly those involved in the coordination of gene expression by Tat. Ultimately, the highly specialised set of molecular interactions identified will provide a framework to further advance our understanding of the mechanisms of HIV-1 proviral gene silencing and activation.</p

Antipredator behaviours of a spider mite in response to cues of dangerous and harmless predators

Prey are known to invest in costly antipredator behaviour when perceiving cues of dangerous, but not of relatively harmless predators. Whereas most studies investigate one type of antipredator behaviour, we studied several types (changes in oviposition, in escape and avoidance behaviour) in the spider mite Tetranychus evansi in response to cues from two predatory mites. The predator Phytoseiulus longipes is considered a dangerous predator for T. evansi, whereas Phytoseiulus macropilis has a low predation rate on this prey, thus is a much less dangerous predator. Spider mite females oviposited less on leaf disc halves with predator cues than on clean disc halves, independent of the predator species. On entire leaf discs, they laid fewer eggs in the presence of cues of the dangerous predator than on clean discs, but not in the presence of cues of the harmless predator. Furthermore, the spider mites escaped more often from discs with cues of the dangerous predator than from discs without predator cues, but they did not escape more from discs with cues of the harmless predator. The spider mites did not avoid plants with conspecifics and predators. We conclude that the spider mites displayed several different antipredator responses to the same predator species, and that some of these antipredator responses were stronger with cues of dangerous predators than with cues of harmless predators

Cicatrização de anastomoses do cólon esquerdo com doença inflamatória: estudo experimental em ratos Healing of the left colon anastomosis with inflammatory bowel disease: experimental study in rats

Complicações relacionadas com a anastomose são descritas com freqüência nas cirurgias para o tratamento da doença inflamatória do cólon. Para conhecer a interferência da inflamação na cicatrização de anastomoses 40 ratos Wistar são utilizados e divididos em 2 grupos. Um deles serviu de controle e no outro induziu-se doença inflamatória com ácido acético 10% por sondagem retal. No sétimo dia procedeu-se à laparotomia em ambos os grupos, colotomia e anastomose término-terminal com pontos separados em plano único. Avaliadas no terceiro e sétimo dias, pode-se verificar que o número de complicações no grupo de animais com doença inflamatória foi maior assim como a mortalidade. As deiscências com peritonite foi a situação mais comum (p=0,0222). A capacidade de suportar pressão, nas anastomoses que evoluíram sem complicações foi menor nestes cólons, porém a diferença quando comparada ao controle não foi significante (p=0,0836). Verificou-se que as anastomoses construídas em cólons com doença inflamatória apresentavam maior concentração de colágeno total, com predomínio de colágeno imaturo (tipo III) (p=0,0000) enquanto que nas feitas em cólons normais predominava colágeno maduro (tipo I) (p=0,0102). Observou-se ainda que a organização do colágeno era menor, no terceiro dia, nas anastomoses com doença inflamatória. Entretando a análise da reação inflamatória ao nível da anastomose foi semelhante nos dois grupos. Estes resultados permitem sugerir que a doença inflamatória leva a aumento do número de deiscências provavelmente pelo atraso da maturação e ordenação do colágeno.<br>Complications related to anastomosis failure are frequently described in the surgery of inflammatory bowel disease. The aim of the present study was to evaluated colonic wound healing in an inflamed bowel. Forty Wistar rats were divided in 2 groups: the control and experimental groups. In the experimental group, colitis was induced by the infusion of 10% acetic acid per rectum to understand how inflammation interferes with the healing process of intestinal. On the7th post-operative day an end to end colonic anastomosis was performed with interrupted suture. On the 3rd and 7th post-operative days, the anastomosis were evaluated. We observed that mortality and the number of complications were greater in the group of animals with inflammatory bowel disease. Dehiscence with peritonitis was the most common complication(p=0.0222). The bursting strength in the colons without leakage was lesser than those in the control group, however this difference was not statistically significant (p=0,0836). It was verified that the anastomosis performed in colons with inflammatory bowel disease showed greater total collagen concentration, with a predominance of immature collagen (type III) (p=0.0000). The mature collagen (type I) was the predominant collagen in the wounds of normal colon. (p=0.0102). On the 3th post-operative day It was also observed that the organization of collagen was poorer on the third day in the colonic wounds with inflammatory bowel disease. Nevertheless, the analysis of the inflammatory reaction at the level of the anastomosis was similar in both groups. These results suggest that inflammatory bowel disease increases the risks of anastomosis dehiscence, probably because of the delay in the maturation and the organisation of collagen