A thiotrophic microbial community in an acidic brine lake in Northern Chile

The endorheic basins of the Northern Chilean Altiplano contain saline lakes and salt flats. Two of the salt flats, Gorbea and Ignorado, have high acidic brines. The causes of the local acidity have been attributed to the occurrence of volcanic native sulfur, the release of sulfuric acid by oxidation, and the low buffering capacity of the rocks in the area. Understanding the microbial community composition and available energy in this pristine ecosystem is relevant in determining the origin of the acidity and in supporting the rationale of conservation policies. Besides, a comparison between similar systems in Australia highlights key microbial components and specific ones associated with geological settings and environmental conditions. Sediment and water samples from the Salar de Gorbea were collected, physicochemical parameters measured and geochemical and molecular biological analyses performed. A low diversity microbial community was observed in brines and sediments dominated by Actinobacteria, Algae, Firmicutes and Proteobacteria. Most of the constituent genera have been reported to be either sulfur oxidizing microorganisms or ones having the potential for sulfur oxidation given available genomic data and information drawn from the literature on cultured relatives. In addition, a link between sulfur oxidation and carbon fixation was observed. In contrast, to acid mine drainage communities, Gorbea microbial diversity is mainly supported by chemolithoheterotrophic, facultative chemolithoautotrophic and oligotrophic sulfur oxidizing populations indicating that microbial activity should also be considered as a causative agent of local acidity.     

Histological characterization of <Emphasis Type="Italic">Passiflora pohlii</Emphasis> Mast. root tips cryopreserved using the V-Cryo-plate technique

Cryopreservation stands out as the main strategy to ensure safe and cost efficient long-term conservation of plant germplasm, especially for biotechnological materials. However, the injuries associated with the procedure may result in structural damage and low recovery rates after cooling. Histological analysis provides useful information on the effects of osmotic dehydration, LN exposure, and recovery conditions on cellular integrity and tissue organization, allowing the determination of the critical steps of the cryopreservation protocol and, thus, the use of optimized treatments. Passiflora pohlii Mast. (Passifloraceae) is a native species from Brazil with potential agronomic interest. Recent studies showed the presence of saponins in its roots, which presented antioxidant activity. The goal of this work was to develop a cryopreservation technique for root tips of in vitro-derived plants of P. pohlii using the V-Cryo-plate technique and to characterize the anatomical alterations that occurred during the successive steps of the protocol. Root tips were excised from in vitro plants and precultured before adhesion to cryo-plates and then treated for different periods with the plant vitrification solutions PVS2 or PVS3. Treatment with PVS2 for 45 min resulted in higher recovery (79%) when compared with PVS3 (43%). The greatest number of adventitious roots per cryopreserved explant was also observed after a 45-min exposure to PVS2. Plasmolysis levels were higher in cortical cells of cryopreserved explants treated with PVS2, while pericycle and central cylinder cells were not damaged after this treatment. Thirty days after rewarming, no plasmolysis could be detected, regardless of the experimental conditions.     

Historical records under the genetic evidence: “Chiriguano” tribe genesis as a test case

Molecular Biology Reports - Historical records suggest that Chiriguano tribe is the result of a genetic admixture event. The process involved the arrival of Guaraní tribesmen descending from...     

Growth and carotenogenesis in eight strains ofDunaliella salina Teodoresco from Chile

A study was made of growth and carotenogenesis in eight strains of the green algaDunaliella salina collected from salt ponds at Salar de Atacama (23° 30′ S; 68° 15′ W) and Antofagasta (23° 39′ S; 70° 24′ W), Chile and kept in unialgal cultures at the Laboratorio Cultivo de Algas, University of Concepcion. The algae were grown in Erdschreiber medium supplemented with 12.5% w/v NaCl, under a continuous photon flux density of approximately 150 μmol m^-2 s^-1 at 25 ± 4 °C without aeration. When growth reached the stationary phase, the amount of NaCl was increased to 25%. Total carotenoid content was measured during the exponential growth phase and 20 days after the addition of salt. Strain CONC-001 (Laguna La Rinconada, Antofagasta) exhibited the highest growth rate (k = 0.32 div d^-1) and the lowest total carotenoid content (7.2 and 13.7 mg l^-1 at 12.5 and 25% NaCl, respectively). Strain CONC-007 (Salar de Atacama) had the lowest growth rate (k = 0.14 div d^-1) and yielded the highest total carotenes per volume unit (23.1 and 35.6 mg 1^-1 at 12.5 and 25% NaCl) and per cell (ca. 42 pg at 25% NaCl). Total carotenoid synthesis did not increase in strains CONC-001 and CONC-006 with the increase of salinity. These strains had the greatest increase of total carotenoid-to-chlorophyll ratio (4.5- and 9.3-fold, respectively). The seven strains from Salar de Atacama had higher total carotenoid contents than the strain from Antofagasta. Cell size also varied. Strain CONC-001 cells were smallest; strain CONC-006 had the largest cells. There was an inverse correlation between maximum cell density and mean cell size.     

A Comparative Study on Nutritional Status and Body Composition of Urban and Rural Schoolchildren from Brandsen District (Argentina)

The purpose of this study was to analyze whether nutritional status and body composition varies according to the environment of residence (urban or rural) of children in the Brandsen district (Argentina). Weight, height, arm circumference and tricipital and subscapular skinfolds were performed in 1368 schoolchildren aged 3 to 14. NHANES III reference was used to estimate nutritional status -underweight, stunting, wasting, overweight, and obesity- and to evaluate body composition -deficit and excess of adipose (DA, EA) and muscular (DM, EM) tissues of the arm-. Central fat distribution (CFD) was estimated using the subscapular-tricipital index. A structured questionnaire was implemented to evaluate socio-environmental characteristics. Nutritional categories based on body size and body composition were compared between urban and rural areas of residence using Chi-squared tests (χ2). The results indicated for the total sample: 1.1% underweight, 6.9% stunting, 0.4% wasting, 12.1% overweight, 9.7% obesity, 22.0% DM, 2.5% EM, 0.1% DA, 17.6% EA, and 8.5% CFD. Significant differences between urban and rural areas were found only for CFD. The socio-environmental analysis showed that while access to public services and housing quality was significantly better in the urban area, a considerable number of city households lived under deficient conditions, lacked health insurance and had low socioeconomic level. Fifty-three percent of the undernourished children had DM without urban-rural significant differences, and none of them showed DA. In the overweight plus obesity group, 62.8% presented EA, 6.4% EM, 4.7% DM, and 22.8% CFD. The highest percentages of DM and CFD were recorded in rural areas (p = 0.00). We conclude that the child population shows the “double burden” of malnutrition. The environment of residence does not promote any differentiation in the nutritional status. Nevertheless, the increment of central adiposity and, in some cases of muscle deficit in rural children, suggests a consumption of unbalanced diet.     

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 Pten^L/L p53^L/L LSL-mTERT^L/L and thymic T-cell lymphoma model G4 Atm^-/- mTERT^ER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.     

Combination of ELISA screening and seroneutralisation tests to expedite Zika virus seroprevalence studies

Here we propose a strategy allowing implementing efficient and practicable large-scale seroepidemiological studies for Zika Virus (ZIKV). It combines screening by a commercial NS1 protein-based Zika IgG ELISA, and confirmation by a cytopathic effect-based virus neutralization test (CPE-based VNT). In post-epidemic samples from Martinique Island blood donors (a population with a dengue seroprevalence above 90%), this strategy allowed reaching specificity and sensitivity values over 98%. The CPE-based VNT consists of recording CPE directly under the optical microscope, which is easy to identify with ZIKV strain H/PF/2013 at day 5 pi. Overall, considered that CPE-based VNT is cost effective and widely automatable, the NS1 protein-based Zika IgG ELISA+CPE-based VNT combination strategy represents a convenient tool to expedite ZIKV seroprevalence studies.     

Incipient colonisation of Lutzomyia longipalpis in the city of Resistencia,province of Chaco,Argentina (2010-2012)

Lutzomyia longipalpis was recorded for the first time in Argentina in 2004, in the province of Formosa. In the following years, the vector spread to the south and west in the country and was recorded in the province of Chaco in 2010. From November 2010-May 2012, captures of Phlebotominae were made in the city of Resistencia and its surroundings, to monitor the spread and possible colonisation of Lu. longipalpis in the province of Chaco. In this monitoring, Lu. longipalpis was absent in urban sampling sites and its presence was restricted to Barrio de los Pescadores. This suggests that the incipient colonisation observed in 2010 was not followed by continuous installation of vector populations and expansion of their spatial distribution as in other urban centres of Argentina.