464 research outputs found

    Smanjeni omjer mineralnog ulja poboljšava prinos blastocista u sustavu mikrojažica i jednozametnim kulturama u poliesterskim mrežama - kratko priopćenje.

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    It is common to apply a mineral oil (MO) overlay to in vitro embryo cultures, even though the use of MO may be harmful to embryo development. To examine whether the volume of MO used in overlays for well-of-the-well (WOW) and polyester mesh (PM) single-embryo culture systems can be reduced, two sets of experiments were performed and the resulting blastocyst formation rates were compared with a conventional group (CG) embryo culture system. In Experiment 1, groups of 20 embryos in 100 μL microdrops of CDM-2 medium were plated in Petri dishes and then were covered with 3.5 mL of MO (resulting in a 1:35 ratio of medium to MO). Groups of 20 embryos were also placed in four-well plates in 400 μL CDM-2 medium per well, and the wells were covered with 400 μL MO (resulting in a 1:1 ratio of medium to MO). In Experiment 2, groups of 20 embryos were plated in four-well plates with 400 μL CDM-2 medium per well, and 1 mL of purified water was added to the center hole of the plate. Two sets of these plates were set up in parallel, and only one of the plates received an additional 400 μL of MO per well (resulting in a 1:1 ratio of medium to MO). A greater percentage of embryos reached the blastocyst stage when they were cultured in a 1:1 ratio of medium to MO, compared with embryos that were cultured in a 1:35 ratio of medium to MO (55 % vs. 41 %, respectively; P<0.01). In addition, a greater percentage of embryos reached the blastocyst stage in the WOW and PM systems that included a 1:1 ratio of medium to MO overlay than the embryos that were cultured in the presence of water in the central hole of a four-well plate without MO (48 % vs. 39 %, respectively; P<0.01). Therefore, a 1:1 ratio of medium to MO was found to improve blastocyst rates in both WOW and PM embryo culture systems.U in vitro kulturama zametaka uobičajeno je primjenjivati sloj mineralnog ulja (MU), iako to može biti štetno za razvoj embrija. U radu su provedeni pokusi s ciljem da se istraži može li se smanjiti količina primijenjenog MU u sustavu mikrojažica i u poliesterskim mrežama za uzgoj pojedinačnog zametka. Pri tome je stopa nastalih blastocista bila uspoređena s konvencionalnim načinom uzgoja zametaka. U prvom pokusu su skupine od 20 zametaka u mikrokapi od 100 μL CDM-2 medija bile stavljene na podlogu u Petrijeve zdjelice i nakon toga prekrivene s 3,5 mL MU (omjer između medija i MU iznosio je 1:35). Skupine od 20 zametaka također su bile smještene u četverodijelne plitice s 400 μL CDM-2 medija po jažici i prekrivene s 400 μL MU (omjer između medija i MU iznosio je 1:1). U drugom pokusu, skupine od 20 zametaka bile su smještene u četverodijelne plitice s 400 μL CDM-2 medija po jažici, u čiji je središnji otvor dodan 1 mL pročišćene vode. Dvije skupine tih plitica bile su istodobno postavljene, a samo u jednu pliticu dodano je 400 μL MU po jažici (omjer između medija i MU iznosio je 1:1). Veći postotak embrija dosegao je stadij blastociste kada je u kulturi bio primijenjen omjer 1:1 između medija i MU. U usporedbi sa zametcima kod kojih je u kulturi bio primijenjen omjer između medija i MU 1:35, razlika je iznosila 55 % prema 41 %, P<0,01. Osim toga, veći postotak zametaka dosegnuo je stadij blastociste u sustavu mikrojažica i poliesterskih mrežica u kojima je omjer između medija i MU bio 1:1, nego što je to bio slučaj kad je kulturi zametaka u središnjoj jažici plitice bila dodana voda (48 % prema 39 %, P<0,01). Utvrđeno je da omjer 1:1 između medija i MU poboljšava stopu razvijenih blastocista u oba sustava

    Vitrification of White-tailed Deer (Odocoileus virginianus) oocytes with sucrose or trehalose for in vitro maturation and fertilization.

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    Objective: Evaluate the White-tailed Deer (WTD) in vitro embryo production (IVP) and oocytes vitrified with Trehalose (TH) or Sucrose (SC). Design/methodology/approach: Total vitrified oocytes were placed into two different groups: TH (n=60) and SC (n=61). Samples were selected and analyzed for viability evaluation TH (n=5) and SC (n=5), nuclear status (NS) TH (n=4) and SC (n=5), Germinal Vesicle (GV), Metaphase I, or not evaluable (NE) after warming. In vitro maturation (IVM) was conducted for 36 h in supplemented TCM-199 medium. Immediately afterwards, oocyte NS was evaluated (n=88) [(GV, MI=immature), (MII=mature)]. In vitro fertilization (IVF) was performed in supplemented TALP medium for 24 h using frozen WTD semen (3x106 sperm/mL), NS was classified [Fertilized (F), Not fertilized (NF), or NE]. Results: After warming, viability for the TH group (n=5) was 60% versus 40% for SC group (n=5), however, oocytes in both groups were immature (GV and MI stage). For IVM, NS evaluations of the TH group (n=38) revealed no maturation versus 2% in the SC group (n=50) (MII stage=matured). IVF evaluations for the TH group (n=10) revealed no fertilization compared to 20% in the SC group (n=5). A statistical difference (p&gt;0.05) was not found between the TH and SC groups. Limitations on study/implications: White-tailed Deer in vitro embryo production is not well documented. Findings/conclusions: Future research with a larger number of WTD oocytes is needed for further evaluation of oocyte vitrification IVP techniques as a model for endangered cervids

    An Intracellular Arrangement of Histoplasma capsulatum Yeast-Aggregates Generates Nuclear Damage to the Cultured Murine Alveolar Macrophages

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    Histoplasma capsulatum is responsible for a human systemic mycosis that primarily affects lung tissue. Macrophages are the major effector cells in humans that respond to the fungus, and the development of respiratory disease depends on the ability of Histoplasma yeast cells to survive and replicate within alveolar macrophages. Therefore, the interaction between macrophages and H. capsulatum is a decisive step in the yeast dissemination into host tissues. Although the role played by components of cell-mediated immunity in the host's defense system and the mechanisms used by the pathogen to evade the host immune response are well understood, knowledge regarding the effects induced by H. capsulatum in host cells at the nuclear level is limited. According to the present findings, H. capsulatum yeast cells display a unique architectural arrangement during the intracellular infection of cultured murine alveolar macrophages, characterized as a formation of aggregates that seem to surround the host cell nucleus, resembling a crown. This extranuclear organization of yeast-aggregates generates damage on the nucleus of the host cell, producing DNA fragmentation and inducing apoptosis, even though the yeast cells are not located inside the nucleus and do not trigger changes in nuclear proteins. The current study highlights a singular intracellular arrangement of H. capsulatum yeast near to the nucleus of infected murine alveolar macrophages that may contribute to the yeast’s persistence under intracellular conditions, since this fungal pathogen may display different strategies to prevent elimination by the host's phagocytic mechanisms

    Black list and Alert list of the Aquatic Invasive Alien Species in the Iberian Peninsula: an action of the LIFE INVASAQUA

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    Resumen del trabajo presentado en VI Congreso Nacional sobre Especies Exóticas Invasoras y I Congreso Ibérico sobre EEI (EEI 2022) celebrado en Navarra del 20 al 23 de abril de 2022.One of the objectives of LIFE INVASQUA project is to develop tools that will be more efficient the Early Warning and Rapid Response (EWRR) framework for Invasive Alien Species in the Iberian Peninsula. Horizon scanning for high-risk IAS is basic in implementing measures to reduce new invasions, developing Alert lists, and to focus effort in the species already established, for instance making a Black list. We developed a trans national horizon scanning exercise focused on inland waters of Spain and Portugal in order to provide a prioritized lists (Black list and Alert list) of aquatic IAS that may pose a threat to aquatic ecosystems and socio economic sectors in the future. We followed a step approach of existing information about IAS (Plants, Freshwater Invertebrates, Estuarine Invertebrates and Vertebrates; 127 established taxa in Black list; 90 non established taxa in Alert list) combining with an expert scoring of prioritized taxa. IAS established in the Iberian aquatic system consistently highlighted as the worst included vertebrates (e.g. Cyprinus carpio, Gambusia holbrooki, Silurus glanis), freshwater and estuarine invertebrates (e.g. Procambarus clarkii, Dreissena polymorpha, Pacifastacus leniusculus, Ficopomatus enigmaticus, Callinectes sapidus, Corbicula fluminea) and plants (e.g. Eichhornia crassipes, Azolla filiculoides, Ludwigia grandiflora). Amongst taxa not yet established (Alert list), expert pointed to Perna viridis, Hydroides dirampha, Dreissena bugensis, Procambarus fallax f. virginallis, Perccottus glenii with higher risk of invasion, ecological and socioeconomic impacts. Over 20.6% of the taxa in the preliminary black list received no votes (no prioritization) by experts, 17.8% in the innitial alert list. Our horizon scanning approach is inclusive of all-taxa, prioritizes both established and emerging biological threats across trans-national scales, and considers not only the ecological impact, but also potential direct economic consequences as well as the manageability of invasive species.This work received funds from the LIFE Programme (LIFE17 GIE/ES/000515)

    Comparative Genomic Analysis of Human Fungal Pathogens Causing Paracoccidioidomycosis

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    Paracoccidioides is a fungal pathogen and the cause of paracoccidioidomycosis, a health-threatening human systemic mycosis endemic to Latin America. Infection by Paracoccidioides, a dimorphic fungus in the order Onygenales, is coupled with a thermally regulated transition from a soil-dwelling filamentous form to a yeast-like pathogenic form. To better understand the genetic basis of growth and pathogenicity in Paracoccidioides, we sequenced the genomes of two strains of Paracoccidioides brasiliensis (Pb03 and Pb18) and one strain of Paracoccidioides lutzii (Pb01). These genomes range in size from 29.1 Mb to 32.9 Mb and encode 7,610 to 8,130 genes. To enable genetic studies, we mapped 94% of the P. brasiliensis Pb18 assembly onto five chromosomes. We characterized gene family content across Onygenales and related fungi, and within Paracoccidioides we found expansions of the fungal-specific kinase family FunK1. Additionally, the Onygenales have lost many genes involved in carbohydrate metabolism and fewer genes involved in protein metabolism, resulting in a higher ratio of proteases to carbohydrate active enzymes in the Onygenales than their relatives. To determine if gene content correlated with growth on different substrates, we screened the non-pathogenic onygenale Uncinocarpus reesii, which has orthologs for 91% of Paracoccidioides metabolic genes, for growth on 190 carbon sources. U. reesii showed growth on a limited range of carbohydrates, primarily basic plant sugars and cell wall components; this suggests that Onygenales, including dimorphic fungi, can degrade cellulosic plant material in the soil. In addition, U. reesii grew on gelatin and a wide range of dipeptides and amino acids, indicating a preference for proteinaceous growth substrates over carbohydrates, which may enable these fungi to also degrade animal biomass. These capabilities for degrading plant and animal substrates suggest a duality in lifestyle that could enable pathogenic species of Onygenales to transfer from soil to animal hosts.National Institute of Allergy and Infectious Diseases (U.S.)National Institutes of Health. Department of Health and Human Services (contract HHSN266200400001C)National Institutes of Health. Department of Health and Human Services(contract HHSN2722009000018C)Brazil. National Council for Scientific and Technological Developmen

    Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

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    The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

    Sloan Digital Sky Survey IV: Mapping the Milky Way, Nearby Galaxies, and the Distant Universe

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    We describe the Sloan Digital Sky Survey IV (SDSS-IV), a project encompassing three major spectroscopic programs. The Apache Point Observatory Galactic Evolution Experiment 2 (APOGEE-2) is observing hundreds of thousands of Milky Way stars at high resolution and high signal-to-noise ratios in the near-infrared. The Mapping Nearby Galaxies at Apache Point Observatory (MaNGA) survey is obtaining spatially resolved spectroscopy for thousands of nearby galaxies (median z0.03z\sim 0.03). The extended Baryon Oscillation Spectroscopic Survey (eBOSS) is mapping the galaxy, quasar, and neutral gas distributions between z0.6z\sim 0.6 and 3.5 to constrain cosmology using baryon acoustic oscillations, redshift space distortions, and the shape of the power spectrum. Within eBOSS, we are conducting two major subprograms: the SPectroscopic IDentification of eROSITA Sources (SPIDERS), investigating X-ray AGNs and galaxies in X-ray clusters, and the Time Domain Spectroscopic Survey (TDSS), obtaining spectra of variable sources. All programs use the 2.5 m Sloan Foundation Telescope at the Apache Point Observatory; observations there began in Summer 2014. APOGEE-2 also operates a second near-infrared spectrograph at the 2.5 m du Pont Telescope at Las Campanas Observatory, with observations beginning in early 2017. Observations at both facilities are scheduled to continue through 2020. In keeping with previous SDSS policy, SDSS-IV provides regularly scheduled public data releases; the first one, Data Release 13, was made available in 2016 July

    The Fourteenth Data Release of the Sloan Digital Sky Survey: First Spectroscopic Data from the Extended Baryon Oscillation Spectroscopic Survey and from the Second Phase of the Apache Point Observatory Galactic Evolution Experiment

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    The fourth generation of the Sloan Digital Sky Survey (SDSS-IV) has been in operation since 2014 July. This paper describes the second data release from this phase, and the 14th from SDSS overall (making this Data Release Fourteen or DR14). This release makes the data taken by SDSS-IV in its first two years of operation (2014–2016 July) public. Like all previous SDSS releases, DR14 is cumulative, including the most recent reductions and calibrations of all data taken by SDSS since the first phase began operations in 2000. New in DR14 is the first public release of data from the extended Baryon Oscillation Spectroscopic Survey; the first data from the second phase of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE-2), including stellar parameter estimates from an innovative data-driven machine-learning algorithm known as "The Cannon"; and almost twice as many data cubes from the Mapping Nearby Galaxies at APO (MaNGA) survey as were in the previous release (N = 2812 in total). This paper describes the location and format of the publicly available data from the SDSS-IV surveys. We provide references to the important technical papers describing how these data have been taken (both targeting and observation details) and processed for scientific use. The SDSS web site (www.sdss.org) has been updated for this release and provides links to data downloads, as well as tutorials and examples of data use. SDSS-IV is planning to continue to collect astronomical data until 2020 and will be followed by SDSS-V
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