Membrane-associated proteins of ejaculated sperm from Morada Nova rams

AbstractThe objective was to describe the profile of membrane proteins from sperm of tropically adapted Morada Nova rams (N = 5). Samples from protein-enriched fractions of ejaculated sperm (containing 400 μg of protein) were separated by two-dimensional electrophoresis and respective maps analyzed using PDQuest software (version 7.3.0; Bio-Rad). Proteins were identified using tandem mass spectrometry. Also, membrane proteins were incubated with antibodies against binder of sperm protein (BSP) 1 and bodhesin 2 (Bdh-2), components of vesicular gland secretion. For membrane proteins of ejaculated sperm, an average of 133 ± 4.6 spots were detected per gel, of which, 107 spots were consistently present on all gels. Sixty-eight spots and 37 proteins were identified using mass spectrometry, corresponding to 71.6% of the intensity of all spots detected. Three major spots identified as ram seminal vesicle protein (RSVP) 14 represented approximately 30% of the intensity of all spots. Two of the most intense spots in the gel reacted against anti-BSP1, at 14 kDa. In addition, four low molecular weight spots reacted with anti-Bdh-2 antibodies. Proteins RSVP and Bdh-2 belong to the BSP and spermadhesin families, respectively, and were previously reported as major components of ram seminal proteins. Additional proteins identified in the sperm membrane two-dimensional maps included alpha-2-heparan sulfate-glycoprotein, plasma glutamate carboxypeptidase, arylsulfatase A, cathelicidin, heat shock protein 70 kDa, angiotensin-converting enzyme, leucine aminopeptidase, and clusterin. Some proteins were present as multiple isoforms, such as tubulin (12), alpha-2-heparan sulfate-glycoprotein (5), ATP synthase (5), Bdh-2 (4) and RSVP14 (3). Based on gene ontology analysis, the most common biological processes associated with the membrane proteins were cellular processes (34%), response to stimulus (14%), and metabolic processes (11%). Binding (37%) and catalytic activity (32%) corresponded to the most frequent molecular functions for those proteins. In conclusion, we identified a diverse cohort of components of membrane proteins in ram sperm. Major proteins previously reported in seminal plasma, such as RSVP14 and Bdh-2, were also extracted from sperm membranes. Knowledge of sperm proteins is crucial for elucidating mechanisms underlying their association with sperm function

Rising Temperature Is a More Important Driver Than Increasing Carbon Dioxide Concentrations in the Trait Responses of Enhalus acoroides Seedlings

Postprin

Host migration and environmental temperature influence avian haemosporidians prevalence: a molecular survey in a Brazilian Atlantic rainforest

Avian haemosporidians are parasites with great capacity to spread to new environments and new hosts, being considered a good model to host-parasite interactions studies. Here, we examine avian haemosporidian parasites in a protected area covered by Restinga vegetation in northeastern Brazil, to test the hypothesis that haemosporidian prevalence is related to individual-level traits (age and breeding season), species-specific traits (diet, foraging strata, period of activity, species body weight, migratory status, and nest shape), and climate factors (temperature and rainfall). We screened DNA from 1,466 birds of 70 species captured monthly from April 2013 to March 2015. We detected an overall prevalence (Plasmodium/Haemoproteus infection) of 22% (44 host species) and parasite’s lineages were identified by mitochondrial cyt b gene. Our results showed that migration can be an important factor predicting the prevalence of Haemoproteus (Parahaemoproteus), but not Plasmodium, in hosts. Besides, the temperature, but not rainfall, seems to predict the prevalence of Plasmodium in this bird community. Neither individual-level traits analyzed nor the other species-specific traits tested were related to the probability of a bird becoming infected by haemosporidians. Our results point the importance of conducting local studies in particular environments to understand the degree of generality of factors impacting parasite prevalence in bird communities. Despite our attempts to find patterns of infection in this bird community, we should be aware that an avian haemosporidian community organization is highly complex and this complexity can be attributed to an intricate net of factors, some of which were not observed in this study and should be evaluated in future studies. We evidence the importance of looking to host-parasite relationships in a more close scale, to assure that some effects may not be obfuscated by differences in host life-history

Araucaria angustifolia (Bertol.) Kuntze extract as a source of phenolic compounds in TPS/PBAT active films

There is growing interest in the development of biodegradable packaging materials containing natural antioxidant extracts. In this sense, the use of extracts obtained from agro-industrial byproducts has proved to be a sustainable alternative. In this study, Pinhão extract, a byproduct of Pinhão (Araucaria angustifolia (Bertol.) Kuntze) seed consumption, was characterized by HPLC-DAD-ESI/MSn, demonstrating the presence of eight phenolic compounds, (+)-catechin and (−)-epicatechin being the most abundant molecules. TPS/PBAT films containing Pinhão extract were produced by blown extrusion and their properties (tensile properties, thermal characteristics and microstructure) were evaluated in order to determine the effect of the presence of extracts. Results suggested that the interaction between the phenolic compounds of the extract and the polymeric matrix caused the reduction in the crystallinity degree, and an increase in the starch glass transition temperature. The presence of Araucaria angustifolia (Bertol.) Kuntze extract significantly (p < 0.05) affected the color and opacity of the film. Regarding water vapor permeation, no significant difference (p > 0.05) was detected. However, the water solubility and the contact angle with water (polar solvent) and diiodomethane (non-polar solvent) significantly changed due to the extract addition. Moreover, the Pinhão extract conferred significant antioxidant capacity to the TPS/PBAT films as determined by DPPH, suggesting that this material can be applied as an active packaging material.The authors thank CNPq (Chamada Universal – MCTI/CNPq no. 14/2014, Processo 447768/2014-0), CAPES (Master’s scholarship) and Fundação Araucária (Programa Universal/ Pesquisa Básica e Aplicada 24/2012, protocolo 7334133700514041013) for the financial support and they also thank Associate Laboratory LSRE-LCM – UID/EQU/50020/2019 – funded by national funds through FCT/MCTES (PIDDAC) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019) and to the national funding by FCT, P.I., through the institutional scientific employment program-contract for L. Barros contracts. The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E.info:eu-repo/semantics/publishedVersio

TPCS/PBAT blown extruded films added with curcumin as a technological approach for active packaging materials

The development of active packaging is a relevant topic demanding the development of films with diverse properties to preserve specific foodstuff. The objective of this work was to obtain extruded TPCS/PBAT films containing curcumin and evaluate it as an active antimicrobial and antioxidant packaging to protect chia oil from oxidative degradation. Morphology, thermal, mechanical, antimicrobial, and antioxidant evaluation of the films were conducted to determine whether the presence of curcumin affected the film’s properties. Infrared Spectroscopy indicated that curcumin addition affected the crosslinking reaction between citric acid and starch, which explains the changes in hydrophilicity and mechanical strength of the films. The incorporation of curcumin conferred antimicrobial activity against Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria, as well as antioxidant activity. Films were tested as chia oil packaging, being verified that they successfully prevented oil degradation under accelerated stability test (60 °C for 7 days), demonstrating the feasibility of using TPCS/PBAT biodegradable films containing curcumin to obtain active packaging materials.Authors thank to CNPq (Chamada Universal– MCTI/CNPq Nº 14/2014, Processo 447768/2014-0), CAPES (Coordenação de Pessoal de Nível Superior Master's scholarship) and Fundação Araucária (Programa Universal/Pesquisa Básica e Aplicada 24/2012, protocolo 7334133700514041013) for the finantial support. This work was financially supported by Associate Laboratory LSRE-LCM (UID/EQU/50020/2019) funded by national funds through FCT/MCTES (PIDDAC), and Foundation for Science and Technology (FCT, Portugal). CIMO (UID/AGR/00690/2019) through FEDER under Program PT2020. To the national funding by FCT, P.I., through the institutional scientific employment program-contract for I.P. Fernandes contract.info:eu-repo/semantics/publishedVersio

Programmable antivirals targeting critical conserved viral RNA secondary structures from influenza A virus and SARS-CoV-2

Influenza A virus’s (IAV’s) frequent genetic changes challenge vaccine strategies and engender resistance to current drugs. We sought to identify conserved and essential RNA secondary structures within IAV’s genome that are predicted to have greater constraints on mutation in response to therapeutic targeting. We identified and genetically validated an RNA structure (packaging stem–loop 2 (PSL2)) that mediates in vitro packaging and in vivo disease and is conserved across all known IAV isolates. A PSL2-targeting locked nucleic acid (LNA), administered 3 d after, or 14 d before, a lethal IAV inoculum provided 100% survival in mice, led to the development of strong immunity to rechallenge with a tenfold lethal inoculum, evaded attempts to select for resistance and retained full potency against neuraminidase inhibitor-resistant virus. Use of an analogous approach to target SARS-CoV-2, prophylactic administration of LNAs specific for highly conserved RNA structures in the viral genome, protected hamsters from efficient transmission of the SARS-CoV-2 USA_WA1/2020 variant. These findings highlight the potential applicability of this approach to any virus of interest via a process we term ‘programmable antivirals’, with implications for antiviral prophylaxis and post-exposure therapy