High genetic variation in marginal fragmented populations at extreme climatic conditions of the Patagonian Cypress Austrocedrus chilensis

Knowledge about current patterns of genetic structure of populations together with the evolutionary history of a species helps to understand and predict the adaptation of populations to future climate change. We assayed variation at nuclear microsatellite markers among peripheral vs. continuous populations of the temperate South American species Austrocedrus chilensis, to investigate the role of historical vs. demographical forces in shaping population genetic structure. This species occurs in continuous populations in the west and central distribution range, but becomes highly fragmented at the eastern limit, which comprised ice-free areas during Quaternary glaciations and has extreme climatic conditions at present times. Bayesian analysis methods identified two contrasting patterns of genetic structure; (I) populations from humid, mesic and peri-glacial regions formed a single deme with relatively low genetic differentiation and high admixture levels whereas (II) a highly heterogeneous genetic structure with low level of admixture was found in the steppe, towards the east and northeast limit of the distribution range. In the steppe, population fragmentation, restricted gene flow and isolation-by-distance were also inferred. In addition, several small steppe populations showed high genetic diversity and divergent gene pools, suggesting that they constitute ancient refuges from pre-Holocene glaciations with just a subgroup of them contributing significantly to post-glacial spread. These results are discussed in relation to patterns of genetic variation found for other temperate species and the contribution of the particular southern Andes topography and climate to post-glacial spread.     

CDKN2A mutations and MC1R variants in Italian patients with single or multiple primary melanoma

We evaluated the contribution of germline CDKN2A mutations and MC1R variants to the development of melanoma in a hospital-based study of single (SPM, n = 398) and multiple primary melanoma (MPM, n = 95). The overall frequency of CDKN2A mutations was 15.2%, and four-fold higher in MPM than in SPM cases (OR = 4.27; 95% CI 2.43-7.53). The likelihood of identifying a CDKN2A mutation increased with family history of melanoma and younger age at diagnosis in MPM cases. Compared to SPM patients, the risk of harboring a CDKN2A mutation rose as the number of primary melanomas increased and was not influenced by family history. The G101W and E27X founder mutations were the most common. Several other mutations (W15X, Q50X, R58X, A68L, A127P and H142R) were detected for the first time in Italian patients. One novel mutation, T77A, was identified. Several non-coding variants with unknown functional significance were also found (5'UTR -25C > T, -21C > T, -67G > C, IVS1 +37G > C); the novel 5'UTR -21C > T variant was not detected in controls. The CDKN2A A148T polymorphism was more frequent in MPM patients than in the control population (15.7% versus 6.6%). Compared to the SPM patients, MPM cases had a 2-fold increased probability of being MC1R variant carriers and a higher probability of carrying two or more variants. No specific association was observed between the type of variant and the number of melanomas, suggesting that the number rather than the type of MC1R variant increases the risk of MPM. We observed no interaction between CDKN2A status and the presence of MC1R variants. The high frequency of CDKN2A mutations in our MPM cases, independent of their family history, is of relevance to genetic counseling and testing in our population.     

Sinorhizobium fredii isolates can be specifically identified by a 260 bp fragment from the nolXWBTUV locus

A pair of primers homologous to the nolXWBTUV locus generated a 260 bp fragment by PCR only in the presence of Sinorhizobium fredii template DNA of different quality. This resulted in a fast and accurate method for the identification of S. fredii either from pure DNA, whole bacterial cells or nodule extracts. By means of two PCR fragments, one specific for S. fredii (260-bp) and the other specific for Bradyrhizobium japonicum (RSalpha), we found that S. fredii strain SMH12 and B. japonicum E109 were equally efficient at developing nodules on soybean plants grown under controlled environmental conditions.     

Prevalence of Toxocara infection in schoolchildren from the Butantã region,São Paulo,Brazil

Visceral larva migrans syndrome by Toxocara affects mainly children between 2 and 5 years of age, it is generally asymptomatic, and the seroprevalence varies from 3 to 86% in different countries. A total of 399 schoolchildren from 14 public schools of the Butant? region, S?o Paulo city, Brazil, were evaluated by Toxocara serology (enzyme-linked immunosorbent assay). Epidemiological data to the Toxocara infection obtained from a protocol were submitted to multiple logistic regression analysis for a risk profile definition. Blood was collected on filter paper by finger puncture, with all samples tested in duplicate. Considering titers > or = 1/160 as positive, the seroprevalence obtained was 38.8%. Among infected children, the mean age was 9.4 years, with a similar distribution between genders. A significant association was observed with the presence of onychophagia, residence with a dirty backyard, living in a slum, previous wheezing episodes, school attended, and family income (p < 0.05). All data, except "living in a slum", were considered to be determinant of a risk profile for the acquisition of Toxocara infection. A monthly income > or = 5 minimum salaries represented a protective factor, although of low relevance. Toxocara eggs were found in at least one of the soil samples obtained from five schools, with high prevalence of Toxocara infections, indicating the frequent soil contamination by this agent.     

Identification of fast and slow growing rhizobia nodulating soybean (Glycine max [L.] Merr) by a multiplex PCR reaction

Two DNA fragments, a 730-bp and a 900-bp fragment, one homologous to host cultivar specificity genes nolBT of Sinorhizobium fredii and the other one homologous to RSalpha, an insertion-like sequence present in Bradyrhizobium japonicum, were generated by polymerase chain reaction (PCR) with two pairs of primers. The amount of each fragment generated by the multiplex PCR was proportional to the amount of template DNA present. The amplification of the 900-bp RSalpha fragment was more sensitive, since it was amplified from a smaller amount of template DNA than the 730-bp nolBT fragment. By running the multiplex reaction in the presence of template DNA isolated from different sources, we confirmed that the reaction can discriminate between S. fredii, Bradyrhizobium japonicum and Sinorhizobium xinjiangensis.     

alpha]-Secretase ADAM10 as well as [alpha]APPs is reduced in platelets and CSF of Alzheimer disease patients

BACKGROUND: Members of membrane-bound disintegrin metalloproteinases (ADAMs) were shown to be capable of cleaving amyloid precursor protein (APP) at the alpha-cleavage site in different cell systems. One of the candidate alpha-secretases identified in this family is ADAM10. The present study addresses the following major questions: 1) Are the levels of an alpha-secretase candidate (i.e., ADAM10) reduced in accessible cells of Alzheimer Disease (AD) patients? 2) Are ADAM10 levels in the peripheral cells of AD patients related to a concomitant decrease in alpha APPs? MATERIALS AND METHODS: Western Blot analysis of ADAM10 is performed on platelet homogenates from 33 sporadic AD patients and on 26 age-matched control subjects. Moreover, the levels of alpha-secretase metabolite (alpha APPs) are tested both in platelets and cerebrospinal fluid (CSF) of the same pool of subjects by means of Western blot with a specific antibody. RESULTS: A significant decrease of platelet ADAM10 levels is observed in patients affected by probable AD when compared to control subjects and this is paralleled by a reduced level of alpha APPs released from platelets. Moreover, in the same pool of AD patients, alpha APPs levels were reduced concomitantly in CSF. CONCLUSIONS: ADAM10 is expressed in platelets. A reduced level of ADAM10 is observed in platelets obtained from AD patients compared to age-matched controls. Further, in the same pool of AD patients, a qualitatively and quantitatively similar decrease in alpha APPs is present both in thrombin-activated platelets and CSF, thus suggesting that alterations of APP processing might occur both in the neuronal compartment and peripheral cells.     

Elevated PTEN levels account for the increased sensitivity of ethanol-exposed cells to tumor necrosis factor-induced cytotoxicity

Tumor necrosis factor alpha (TNF) is known to be one of the primary causative cytokines inflicting the characteristic damage to hepatocytes seen in alcoholic liver disease. TNF activates both cell survival and death-inducing signaling pathways. The balance between these two prongs determines the fate of the cell and the onset of disease. Ethanol exposure has been shown to alter mitochondrial function, decreasing their threshold for injury. Importantly, mitochondrial injury is a necessary end point of TNF-induced cell killing. It has been shown that ethanol exposure increases the sensitivity of hepatocytes and HepG2E47 cells to TNF-mediated death. The cumulative and terminal effect of the increased sensitivity to TNF caused by ethanol is an induction of a mitochondrial permeability transition. TNF brings about the mitochondrial permeability transition in ethanol-exposed cells due to amplification in the activity of the p38 stress kinase and a diminution in the activity of the antiapoptotic Akt/PKB kinase. The present report identifies an increase of PTEN expression in ethanol-exposed cells as the main causative factor in altering the balance between prosurvival and prodeath signals initiated by TNF. Suppression of the elevated PTEN levels found in ethanol-exposed HepG2E47 cells through the use of RNA interference reversed the ethanol-induced alterations to TNF signaling, resulting in a preservation of mitochondrial function and cell viability.     

Microgravity-induced alterations in cultured testicular cells

Cultured STe cells (2n karyotype) from swine testis were submitted to simulated microgravity using a 3D Random Positioning Machine for 5 min., 15 min., 30 min., 1 h and 23 h. Sample processing included: histological characterization of cell types, immunohistochemical identification of (i) microtubules (a-tubulin), (ii) alkaline phosphates, (iii) 3 beta-hydroxy-steroid-dehydrogenase (3?-HSDH), and histochemical lipid analyses. After 5 min. simulated microgravity a slight microtubule disorganisation occurred, which increased dramatically with increasing microgravity duration. After 23 h microtubule arrays were completely disrupted. 3 beta-HSDH immunostaining was detectable only in one cell type: under control conditions and 5 min. into microgravity immunoreactivity was strong, but completely disappeared thereafter. Immunostaining intensity for alkaline phosphates, a good marker for myoid cells, decreased after 15 min. in microgravity.     

Analytical SAR computation in a multilayer elliptic cylinder: the near-field line-current radiation case

In this paper, a previously proposed analytical procedure for the computation of the specific absorption rate (SAR) inside a biological elliptic cylinder model is extended to the case in which the body is illuminated under near-field conditions. The elliptic model is made up of layers of different biological tissues and the source is constituted by a line-current distribution. The recursive procedure in which the field is expressed in terms of Mathieu function is modified to express the incident electromagnetic wave produced by the line current. The new procedure makes it possible to check and validate numerical solutions obtained by accurate numerical techniques for SAR prediction, under more realistic illumination condition.