Formation of an RNase A derivative containing an aminosuccinyl residue in place of asparagine 67

At acidic pH, Asp67 and beta-Asp67 (beta-Asp: isoaspartic acid residue) derivatives of RNase A, obtained by selective deamidation of the parent enzyme, spontaneously produces a new derivative containing an aminosuccinyl residue (Asu). The overall secondary structure of the protein chain does not change as a consequence of this substitution, while the catalytic activity on RNA is reduced to about 25%. The pH dependence of the first-order rate constants for the Asu formation has a bell-shaped profile, the maximum being close to the pK(a) of the aspartic acid side chains. Moreover, the values of the rate constants are of the same magnitude of those measured for Asp-containing peptides whose sequence mimics the Asu formation site of the enzyme. This feature indicates that Asp67 and beta-Asp67 residues in the deamidated RNase A derivatives are sited in a region flexible enough to permit the cyclization of the carboxylic side chain to succinimide ring. These results are discussed at the light on to the three-dimensional structure and the thermodynamic stability of the aspartic acid derivatives of RNase A.     

Zinc transport and metallothionein secretion in the intestinal human cell line Caco-2

Caco-2, a human cell line, displays several biochemical and morphological characteristics of differentiated enterocytes. Among these is the ability to transport zinc from the apical to the basal compartment. This process was enhanced following exposure by the apical compartment to increasing concentrations of the metal. High pressure liquid chromatography fractionation of the media obtained from cells labeled with radioactive zinc showed that metallothioneins (MTs), small metal-binding, cysteine-rich proteins), were present in the apical and basal media of controls as well as in cells grown in the presence of high concentrations of zinc. Following exposure to the metal, the levels of Zn-MTs in the apical medium increased, while in the basal compartment the greatest part of zinc appeared in a free form with minor changes in the levels of basal MTs. Metabolic labeling experiments with radioactive cysteine confirmed the apical secretion of MTs. A stable transfectant clone of Caco-2 cells (CL11) was selected for its ability to express constitutively high levels of the mouse metallothionein I protein. This cell line showed an enhanced transport of the metal following exposure to high concentrations of zinc and a constitutive secretion of the mouse metallothionein I protein in the apical compartment. Together, these findings strongly support the hypothesis of a functional role between the biosynthesis and secretion of MTs and the transport of zinc in intestinal cells.     

Adenosine receptors are involved in the control of acute naloxone-precipitated withdrawal: in vitro evidence

The effects exerted by adenosine A1 and A2 receptor agonists and antagonists on the acute opiate withdrawal induced by morphine were investigated in vitro. Following a 4 min in vitro exposure to morphine, the guinea-pig isolated ileum exhibited a strong contracture after the addition of naloxone. The P1 adenosine receptor agonist, adenosine, was able to reduce dose-dependently naloxone-precipitaded withdrawal. The same effect was induced by the adenosine A1 receptor agonist, N6-Cyclopentyladenosine (CPA) whereas the selective adenosine A2A receptor agonist CGS 21680 increased the naloxone-precipitated withdrawal phenomenon. Dipyridamole, a blocker of adenosine reuptake, induced a significant reduction of morphine dependence. Caffeine, an adenosine receptor antagonist, significantly increased the naloxone-precipitated withdrawal effect in a concentration dependent manner. The same effect was observed with 8-phenyltheophylline (8PT), an A1 adenosine receptor antagonist, whereas 3,7-dimethyl-1-propargylxanthine (DMPX), an A2 adenosine receptor antagonist, reduced the naloxone-precipitated withdrawal phenomenon. The results of our experiments indicate that both A1 and A2 adenosine receptor agonists and antagonists are able to influence opiate withdrawal in vitro, suggesting an important functional interaction between the adenosine receptors and opioid withdrawal.     

Synthesis and properties of L-cysteinyl-L-cysteine disulfides

Oligomeric cyclic disulfides, obtained by mild oxidation of the fully protected dipeptide L -cysteinyl-L -cysteine, have been isolated by gel and thin-layer chromatography. Polymeric material was recycled by a thiol–disulfide exchange-reaction performed at basic pH. Spectroscopic investigations of the monomer and the two dimers indicate that conformers characterized by dihedral angles about the S? S bond close to ±90° are preferred. Moreover, chiroptical and ¹H-nmr data for these compounds suggest higher mobility for the two dimers. The antiparallel dimeric disulfide can be considered a model compound for the hinge region formed at the subunit interface of the bovine seminal ribonuclease, a dimeric enzyme showing a complex kinetic behavior.     

Predation of army ants by Toppin’s titi monkey, <Emphasis Type="Italic">Plecturocebus toppini</Emphasis> Thomas 1914 (Primates: Pitheciidae), in an urban forest fragment in eastern Acre,Brazil

The predation of army ants (Eciton rapax) was recorded during an observational study of the feeding behavior of a group of titi monkeys (Plecturocebus toppini) in an urban fragment of forest in Acre, Brazil. During one observed event, the group’s adult female used its tail to retrieve ants, a type of behavior not observed previously in this genus. All incidents of on-forest floor foraging occurred during the dry season, when fruit was least abundant in the forest, while on other occasions, the ants were captured from tree branches and leaves. The observation of predation and ground-level foraging recorded in this study reinforce the adaptive capacity of P. toppini for survival in fragmented forests, and this was also the first record of the predation of army ants by this titi species.     

On the teaching of ecological restoration in Brazil: an analysis of postgraduate courses

Several ambitious restoration targets have been set for recovering degraded ecosystems in Brazil (21 million hectares with a legal deficit of native vegetation). The development of scientific knowledge and skilled professionals is necessary to meet this demand and is fundamental for achieving success in medium‐ and long‐term restoration. In this study, we describe the availability and profile of postgraduate courses on ecological restoration in Brazil. We surveyed 272 postgraduate programs, comprising six specific areas of concentration (forestry, soil sciences, botany, environmental engineering, ecology, and environmental science). We analyzed all selected courses in terms of location, approach (economic, social, and legal), and restoration techniques. Only 27% of the postgraduate programs analyzed offer ecological restoration courses, and 49% of these are offered in the southeast region of the country. Regions covered by nonforest ecosystems, such as Pampas, Caatinga, and Pantanal, were poorly represented. Legal aspects were considered in 51% of the courses, whereas social and economic approaches were only considered in 2.7 and 11% of the courses, respectively. Seedling planting (85%) and nucleation (68%) were the most frequently cited techniques of active restoration, and passive restoration was mentioned in 63% of the courses. Our findings highlight the need for increasing postgraduate courses in areas with nonforest ecosystems, especially due to a lack of knowledge about restoration of these areas. The incorporation of economic and social approaches is highly recommended, considering the importance of these attributes in achieving success in large‐scale restoration initiatives.     

Cellular and subcellular localization of uncoupling protein 2 in the human kidney

The uncoupling protein-2 (UCP2) is an anion transporter that plays a key role in the control of intracellular oxidative stress. In animal models UCP2 downregulation has several pathological sequelae, particularly affecting the vasculature and the kidney. Specifically, in these models kidney damage is highly favored in the absence of UCP2 in the context of experimental hypertension. Confirmations of these data in humans awaits further information, as no data are yet available concerning the cell-type and subcellular expression in the human kidney. In the present study, we aimed to characterize the UCP2 protein distribution in human kidney biopsies. In humans UCP2 is mainly localized in proximal convoluted tubule cells, with an intracytoplasmic punctate staining. UCP2 positive puncta are often localized at the interface between the endoplasmic reticulum and the mitochondria. Glomerular structures do not express UCP2 at detectable levels. The expression of UCP2 in proximal tubular cells may explain their relative propensity to damage in pathological conditions including the hypertensive disease.     

El fenómeno de las caídas en residencias e instituciones: revisión del Grupo de Trabajo de Osteoporosis,Caídas y Fracturas de la Sociedad Española de Geriatría y Gerontología (GCOF-SEGG)

The Workshop on Osteoporosis, Falls and Fractures (GCOF) of The Spanish Geriatrics and Gerontology Society (SEGG) formed a committee in order to review the state of the art on the detection, risk factors and assessment tools for falls, and intervention protocols when falls occurs in nursing homes, long-term hospitals or medium-stay units. The different patient profiles are described in order to make a comprehensive approach to this heterogeneous topic and population, offering a risk classification and specific advice according to these categories.