Ancient Microbes from Halite Fluid Inclusions: Optimized Surface Sterilization and DNA Extraction

Fluid inclusions in evaporite minerals (halite, gypsum, etc.) potentially preserve genetic records of microbial diversity and changing environmental conditions of Earth's hydrosphere for nearly one billion years. Here we describe a robust protocol for surface sterilization and retrieval of DNA from fluid inclusions in halite that, unlike previously published methods, guarantees removal of potentially contaminating surface-bound DNA. The protocol involves microscopic visualization of cell structures, deliberate surface contamination followed by surface sterilization with acid and bleach washes, and DNA extraction using Amicon centrifugal filters. Methods were verified on halite crystals of four different ages from Saline Valley, California (modern, 36 ka, 64 ka, and 150 ka), with retrieval of algal and archaeal DNA, and characterization of the algal community using ITS1 sequences. The protocol we developed opens up new avenues for study of ancient microbial ecosystems in fluid inclusions, understanding microbial evolution across geological time, and investigating the antiquity of life on earth and other parts of the solar system

Quality of life of adult retinoblastoma survivors in the Netherlands

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

Competitive Tendering In The Netherlands: Central Planning Or Functional Specifications?

Institute of Transport and Logistics Studies. Faculty of Economics and Business. The University of Sydne

Conserved Genes Act as Modifiers of Invertebrate SMN Loss of Function Defects

Spinal Muscular Atrophy (SMA) is caused by diminished function of the Survival of Motor Neuron (SMN) protein, but the molecular pathways critical for SMA pathology remain elusive. We have used genetic approaches in invertebrate models to identify conserved SMN loss of function modifier genes. Drosophila melanogaster and Caenorhabditis elegans each have a single gene encoding a protein orthologous to human SMN; diminished function of these invertebrate genes causes lethality and neuromuscular defects. To find genes that modulate SMN function defects across species, two approaches were used. First, a genome-wide RNAi screen for C. elegans SMN modifier genes was undertaken, yielding four genes. Second, we tested the conservation of modifier gene function across species; genes identified in one invertebrate model were tested for function in the other invertebrate model. Drosophila orthologs of two genes, which were identified originally in C. elegans, modified Drosophila SMN loss of function defects. C. elegans orthologs of twelve genes, which were originally identified in a previous Drosophila screen, modified C. elegans SMN loss of function defects. Bioinformatic analysis of the conserved, cross-species, modifier genes suggests that conserved cellular pathways, specifically endocytosis and mRNA regulation, act as critical genetic modifiers of SMN loss of function defects across species

EFFECT OF FLUID INTAKE ON CHANGING BLOOD VOLUME IN HEALTHY MALES

Matthew A. Tucker1, Cory L. Butts1, Nicole E. Moyen1, Evan C. Johnson1,2, J. D. Adams1, Alf Z. Satterfield1, Ashley Six1 & Matthew S. Ganio1. Human Performance Laboratory, University of Arkansas, Fayetteville, Arkansas; 2Department of Kinesiology and Health, University of Wyoming, Laramie, Wyoming. Given that euhydration should represent an optimal total body water (TBW) level and there is a clear relationship between TBW and BV, it is important to understand how fluid intake may change BV. It is well documented that altered fluid intake changes urinary and circulatory markers of hydration status; however, the degree to which changes in traditional indices of hydration status are simultaneously reflected in measurements of blood volume (BV) remains unknown. PURPOSE: To investigate changes in hydration status and BV in response to 24 h of controlled fluid intake. METHODS: Seventeen healthy male subjects (age 24 ± 3 y, mass 84.4 ± 8.4 kg) were provided food and water over 24 h that had a total water volume of 60 ml/kg fat-free mass. Plasma volume (PV) and BV (via CO rebreathing) along with serum and urine osmolality (Sosm and Uosm, respectively) were measured pre- and post-intervention. Urine was collected over the 24-h period and analyzed for volume (24-Uvol) and osmolality (24-Uosm). Based on BV responses to the intervention, subjects were post-hoc assigned to groups in which BV had either increased (BVInc: n = 9, 362 ± 136 ml) or decreased (BVDec: n = 8, -493 ± 247 ml; p \u3c 0.001). RESULTS: Total fluid intake was not different between groups (3930 ± 322 vs. 3883 ± 468 ml; p = 0.813, for BVInc and BVDec, respectively). The groups started the hydration protocol with similar Sosm and Uosm (both p \u3e 0.05). However, the BVInc group started with a lower PV (3517 ± 481 ml) and BV (6228 ± 653 ml) versus the BVDec group (4005 ± 345 ml and 7081 ± 644 ml, respectively; p \u3c 0.05). With the fluid intervention, BVInc had an increase in BV (362 ± 136 ml) and PV (268 ± 84 ml) while BVDec had a decrease in BV (-493 ± 247 ml) and PV (-272 ± 110 ml). This lead to similar PV (3875 ± 512 and 3734 ± 386 ml, for BVInc and BVDec; p \u3e 0.05) and BV (6590 ± 669 and 6588 ± 661 ml, p \u3e 0.05) at the end of the 24-h fluid intervention. Interestingly, with 24-h of prescribed fluid, the change in Sosm (-1 ± 2 vs. -2 ± 2 mOsm/kg) and Uosm (-133 ± 190 vs. -33 ± 316 mOsm/kg), along with 24-Uvol (2819 ± 978 vs. 3218 ± 711 ml) or 24-Uosm (377 ± 177 vs. 324 ± 81 mOsm/kg) were similar between BVInc and BVDec (all p \u3e 0.05). CONCLUSION: Despite no significant differences in traditional indices of hydration status prior to the intervention, two groups of subjects’ blood volume responded in opposite fashion to the same volume of fluid over 24 h. Interestingly, the changes in blood volume were not reflected in changes in traditional hydration biomarkers. This may suggest that, while they appeared to begin similarly hydrated, the BVInc group had below optimal TBW level, as evidenced by the retention of fluid (and subsequent increase in BV) during a period of prescribed fluid intake

Transtorno de oposição e desafio e transtorno de conduta: os desfechos no TDAH em adultos Oppositional defiant disorder and conduct disorder: their outcomes into adulthood

Os autores examinam a influência dos transtornos de oposição e desafio (TOD), de conduta (TC) e de personalidade anti-social (TPAS) ao longo da vida do indivíduo com TDAH. Os principais achados mostram que o TDAH é modulado por essas comorbidades e que seu prognóstico é modificado dependendo da presença ou não desses transtornos. O transtorno de oposição e desafio intensificaria as características de impulsividade e isolacionismo do TDAH, porém não acarretaria em um aumento na incidência de TPAS na vida adulta. Já o TC associado ao TDAH implica um aumento significativo na impulsividade e agressividade, estando associado significativamente a TPAS e um pior prognóstico. A diferenciação entre os diferentes transtornos e seu correto diagnóstico é essencial para o tratamento adequado do TDAH. Futuros estudos precisam determinar se o tratamento do TDAH produziria uma mudança significativa no prognóstico desse grupo de pacientes.<br>The authors examine the influence of oppositional defiant disorder (ODD), conduct disorder (CD) and anti-social personality disorder (ASPD) on attention deficit/hyperactivity disorder (ADHD) across life span. The findings showed that ADHD is modulated by this comorbidities and ADHD prognosis is modified depending on the presence or the absence of those disorders. ODD intensifies ADHD impulsivity and isolationism, but does not lead to an increase in the prevalence of ASPD in adulthood. Otherwise, CD associated with ADHD increases significantly the levels of impulsivity and aggressiveness, is associated with ASPD and a poor outcome. The appropriate approach to ADHD must be based on the correct diagnosis of different comorbidities to predict the outcomes. Further studies are needed to investigate if the treatment of ADHD can produce a significant improvement on the outcomes of this group of patients