Adverse effects of tempol on hidrosaline balance in rats with acute sodium overload

We studied the effects of tempol, an oxygen radical scavenger, on hydrosaline balance in rats with acute sodium overload. Male rats with free access to water were injected with isotonic (control group) or hypertonic saline solution (0.80 mol/l NaCl) either alone (Na group) or with tempol (Na-T group). Hydrosaline balance was determined during a 90 min experimental period. Protein expressions of aquaporin 1 (AQP1), aquaporin 2 (AQP2), angiotensin II (Ang II) and endothelial nitric oxide synthase (eNOS) were measured in renal tissue. Water intake, creatinine clearance, diuresis and natriuresis increased in the Na group. Under conditions of sodium overload, tempol increased plasma sodium and protein levels and increased diuresis, natriuresis and sodium excretion. Tempol also decreased water intake without affecting creatinine clearance. AQP1 and eNOS were increased and Ang II decreased in the renal cortex of the Na group, whereas AQP2 was increased in the renal medulla. Nonglycosylated AQP1 and eNOS were increased further in the renal cortex of the Na-T group, whereas AQP2 was decreased in the renal medulla and was localized mainly in the cell membrane. Moreover, p47-phox immunostaining was increased in the hypothalamus of Na group, and this increase was prevented by tempol. Our findings suggest that tempol causes hypernatremia after acute sodium overload by inhibiting the thirst mechanism and facilitating diuresis, despite increasing renal eNOS expression and natriuresis.     

Arsenic stress affects the expression profile of genes of 14-3-3 proteins in the shoot of mycorrhiza colonized rice

The intimate association between the arbuscular mycorrhizal fungi and host plants helps the latter in phosphate acquisition in exchange of carbohydrates and in enhanced stress tolerance. Similarly, the ubiquitous 14-3-3 protein family is known to be a major regulator of plant metabolism and stress responses. However, the involvement of mycorrhiza and plant 14-3-3 proteins interaction in plant response to environmental stimuli, such as arsenic (As) stress, is yet unknown. In this study, we analysed the impact of the As stress on the expression profile of 14-3-3 genes in the shoot of mycorrhiza colonized rice (Oryza sativa) plants. Ten day old rice seedlings were kept for 45 days for mycorrhizal colonisation (10 g inoculum per 120 g soilrite) and were then subjected to 12.5 µM arsenate [As(V)] exposure for 1 and 3 days, in hydroponics. Arsenate stress resulted in significant change in expression of 14-3-3 protein genes in non-colonized and mycorrhiza colonized rice plants which indicated As mediated effects on 14-3-3 proteins as well as interactive impact of mycorrhiza colonization. Indeed, mycorrhiza colonization itself induced up-regulation of all 14-3-3 genes in the absence of As stress. The results thus indicate that 14-3-3 proteins might be involved in As stress signalling and the mycorrhiza induced As stress response of the rice plants.     

In vitro cell-based assays for evaluation of antioxidant potential of plant-derived products

Several plant-derived compounds have been screened by antioxidant assays, but many of these results are questionable, since they do not evaluate the pharmacologic parameters. In fact, the development of better antioxidants stills a great challenge. In vitro cell-based assays have been employed to assess the antioxidant effect of various compounds at subcellular level. Cell-based assays can also reveal compounds able to enhance the antioxidant pathways, but without direct radical scavenging action (which could not be detected by traditional assays). These methodologies are general of easy implementation and reproducible making them suitable for the early stages of drug discovery. Hydrogen peroxide, a nonradical derivative of oxygen, can be employed as an oxidative agent in these assays due its biochemical properties (presence of all biological systems, solubility) and capacity to induce cell death. Truthfully, if their limitations are understood (such as difference on cell metabolism when in in vitro conditions), these cell-based assays can provide useful information about the pathways involved in the protective effects of phytochemicals against cell death induced by oxidative stress, which can be exploited to develop new therapeutic approaches.     

Our experience with combined procedures in aesthetic plastic surgery

The instep flap needs neither muscle nor a transposition base for survival or innervation. It can be transposed as an island fasciocutaneous flap either on the medial or lateral plantar neurovascular bundles or both, and it can be transferred also as a free flap from the opposite foot. Four cases demonstrating the use of the flap as an island and free flap are presented with follow-up ranging from 1 to 2 years. The absence of muscle in the flap provides greater stability of the heel reconstruction and results in a lesser secondary defect. Sensation in the flaps is diminished but adequate for long-term function, but hyperkeratotic reaction remains an unpredictable problem. The ability to transfer the flap as a free transfer widens the scope of the flap to reconstruct both heel and forefoot defects where local instep tissue or vascularity are inadequate for local reconstruction. The secondary defect, particularly when no muscle is included in the flap, has been minimal.     

Functional characterization of cloned intrahepatic, hepatitis B virus nucleoprotein-specific helper T cell lines

We have previously reported the establishment and preliminary characterization of polyclonal hepatitis B virus (HBV) nucleoprotein (HBcAg)-specific CD4+ and CD8+ T cell lines derived from the hepatic lymphomononuclear cell infiltrate of several patients with chronic active hepatitis B. The isolated subsets from these lines were specifically activated by HBcAg and displayed antigen-specific help and suppression with respect to proliferation of the alternate subset. One of these lines was recently cloned by limiting dilution, and four HBcAg-specific CD3+ CD4+ CD8-DR+ T cell lines were produced that had a 95.3% likelihood of monoclonality. Antigenic specificity was confirmed by dose-dependent, HLA class II (DR)-restricted proliferation in response to recombinant and human serum-derived HBcAg and the lack of proliferation to HBV envelope antigens (HBsAg and pre-S(2)Ag). All cloned lines were interleukin 2 dependent, produced interferon-gamma in an antigen-specific manner, and provided antigen-specific help to autologous B cells with respect to anti-HBc production. We conclude that HBcAg-specific, HLA-class II restricted helper T cells capable of inducing antigen-specific functional responses by autologous B lymphocytes and T lymphocytes are present at the site of viral antigen synthesis and hepatocellular injury in HBV infection.     

Type II restriction endonucleases from Helicobacter pylori include an enzyme with a novel recognition sequence

Type II restriction endonuclease activities of Helicobacter pylori strain Roberts and of the type strain H. pylori NCTC 11637 were detected and analysed by conventional techniques. The endonucleases were partially purified, their optima for activity and their recognition and cleavage sites were determined. H. pylori (Roberts) contained at least two enzymes: HpyBI was an isoschizomer of RsaI (GT/AC) and HpyBII was of a novel specificity (GTN/NAC). H. pylori NCTC 11637 was found to contain an isoschizomer of EcoRV (HpyCI: GAT/ATC) and at least one other enzyme which was too unstable to characterise.