Biosensor capability of the endometrium is mediated in part, by altered miRNA cargo from conceptus-derived extracellular vesicles.

We tested the hypothesis that the biosensor capability of the endometrium is mediated in part, by the effect of different cargo contained in the extracellular vesicles secreted by the conceptus during the peri-implantation period of pregnancy. We transferred Bos taurus taurus embryos of different origin, in vivo (high developmental potential (IV)), in vitro (intermediate developmental potential (IVF)), or cloned (low developmental potential (NT)), into Bos taurus indicus recipients. Extracellular vesicles (EVs) recovered from Day 16 conceptus-conditioned medium were characterized and their microRNA (miRNA) cargo sequenced alongside RNA sequencing of their respective endometria. There were substantial differences in the endometrial response to in vivo versus in vitro and in vivo versus cloned conceptuses (1153 and 334DEGs respectively) with limited differences between in vitro Vs cloned conceptuses (36 DEGs). The miRNA cargo contained in conceptus-derived EVs was similar between all three groups (426 miRNA in common). Only 8 miRNAs were different between in vivo and cloned conceptuses, while only 6 miRNAs were different between in vivo and in vitro-derived conceptuses. Treatment of endometrial epithelial cells with mimic or inhibitors for miR-128 and miR-1298 changed the proteomic content of target cells (96 and 85, respectively) of which mRNAs are altered in the endometrium in vivo (PLXDC2, COPG1, HSPA12A, MCM5, TBL1XR1, and TTF). In conclusion, we have determined that the biosensor capability of the endometrium is mediated in part, by its response to different EVs miRNA cargo produced by the conceptus during the peri-implantation period of pregnancy.

Extracellular vesicles-coupled miRNAs from oviduct and uterus modulate signaling pathways related to lipid metabolism and bovine early embryo development.

BACKGROUND: Extracellular vesicles (EVs) present in oviductal (OF) and uterine fluid (UF) have been shown to enhance bovine embryo quality during in vitro culture by reducing lipid contents and modulating lipid metabolism-related genes (LMGs), while also influencing cell proliferation, suggesting their involvement on the regulation of different biological pathways. The regulation of signaling pathways related to cell differentiation, proliferation, and metabolism is crucial for early embryo development and can determine the success or failure of the pregnancy. Bioactive molecules within EVs in maternal reproductive fluids, such as microRNAs (miRNAs), may contribute to this regulatory process as they modulate gene expression through post-transcriptional mechanisms. RESULTS: From the 20 differentially expressed miRNAs, 19 up-regulated in UF-EVs (bta-miR-134, bta-miR-151-3p, bta-miR-155, bta-miR-188, bta-miR-181b, bta-miR-181d, bta-miR-224, bta-miR-23b-3p, bta-miR-24-3p, bta-miR-27a-3p, bta-miR-29a, bta-miR-324, bta-miR-326, bta-miR-345-3p, bta-miR-410, bta-miR-652, bta-miR-677, bta-miR-873 and bta-miR-708) and one (bta-miR-148b) in OF-EVs. These miRNAs were predicted to modulate several pathways such as Wnt, Hippo, MAPK, and lipid metabolism and degradation. Differences in miRNAs found in OF-EVs from the early luteal phase and UF-EVs from mid-luteal phase may reflect different environments to meet the changing needs of the embryo. Additionally, miRNAs may be involved, particularly in the uterus, in the regulation of embryo lipid metabolism, immune system, and implantation. This study evaluated miRNA cargo in OF-EVs from the early luteal phase and UF-EVs from the mid-luteal phase, coinciding with embryo transit within oviduct and uterus in vivo, and its possible influence on LMGs and signaling pathways crucial for early embryo development. A total of 333 miRNAs were detected, with 11 exclusive to OF, 59 to UF, and 263 were common between both groups. CONCLUSIONS: Our study suggests that miRNAs within OF- and UF-EVs could modulate bovine embryo development and quality, providing insights into the intricate maternal-embryonic communication that might be involved in modulating lipid metabolism, immune response, and implantation during early pregnancy.

Can We Predict Kidney Graft Function and Graft Survival Using Hypothermic Machine Perfusion Parameters From Donors After Circulatory Death?

Background: Hypothermic machine perfusion (HMP) reduces renal injury in donation after circulatory death donors with a high Kidney Donor Profile Index (KDPI). This study aims to characterize the correlation between KDPI, HMP parameters, and donor vitals during the withdrawal period in predicting short- and long-term graft outcomes. Methods: ANOVA with Tukey's honestly significant difference tests compared the relationship between average flow, average resistance, peak resistance, flow slope, and resistance slope on day 30, 1-y, and 3-y eGFR, and days of delayed graft function. Graft and recipient survival rates were assessed using Kaplan-Meier analysis. Results: The data for 72 grafts were suitable for analysis. Kidneys with KDPI >50% had a significantly higher day 30, and 1-y posttransplant eGFR, if HMP average flow was >150 mL/min, or the average resistance was <0.15 mm Hg/mL/min, compared with kidneys with also KDPI >50% but had not achieved the same pump parameters. There were no significant differences in the Kaplan-Meier analysis, considering recipient or graft survival, regardless of the KPDI score with 3- or 5-y outcomes. Conclusions: Use of average resistance and average flow from a HMP, in conjunction with KDPI, may be predictive of the short- and long-term function of donation after circulatory death kidney transplants.

Observation of magnetic vortex configuration in non-stoichiometric Fe3O4 nanospheres.

Theoretical and micromagnetic simulation studies of magnetic nanospheres with vortex configurations suggest that such nanostructured materials have technological advantages over conventional nanosystems for applications based on high-power-rate absorption and subsequent emission. However, full experimental evidence of magnetic vortex configurations in spheres of submicrometer size is still lacking. Here, we report the microwave irradiation fabrication of Fe3O4 nanospheres and establish their magnetic vortex configuration based on experimental results, theoretical analysis, and micromagnetic simulations. Detailed magnetic and electrical measurements, together with Mössbauer spectroscopy data, provide evidence of a loss of stoichiometry in vortex nanospheres owing to the presence of a surface oxide layer, defects, and a higher concentration of cation vacancies. The results indicate that the magnetic vortex spin configuration can be established in bulk spherical magnetite materials. This study provides crucial information that can aid the synthesis of magnetic nanospheres with magnetically tailored properties; consequently, they may be promising candidates for future technological applications based on three-dimensional magnetic vortex structures.

A Prospective Study of the Effect of Gastroduodenal Artery Reconstruction on Duodenal Oxygenation and Enzyme Content After Pancreas Transplantation.

BACKGROUND: Whole pancreas transplantation provides durable glycemic control and can improve survival rate; however, it can carry an increased risk of surgical complications. One devastating complication is a duodenal leak at the site of enteroenteric anastomosis. The gastroduodenal artery (GDA) supplies blood to the donor duodenum and pancreas but is commonly ligated during procurement. Since we have not had expressive changes in pancreatic back table surgical techniques in the recent decades, we hypothesized whether back table GDA reconstruction, improving perfusion of the donor duodenum and head of the pancreas, could lead to fewer surgical complications in simultaneous pancreas-kidney (SPK) transplants. MATERIAL AND METHODS: Between 2017 and 2021, we evaluated demographic information, postoperative complications, intraoperative donor duodenum, recipient bowel O2 tissue saturation, and patient morbidity through the Comprehensive Complication Index (CCI®). RESULTS: A total of 26 patients were included: 13 underwent GDA reconstruction (GDA-R), and 13 had GDA ligation (GDA-L). There were no pancreatic leaks in the GR group compared to 38% (5/13) in the GDA-L group (p = 0.03913). Intraoperative tissue oxygen saturation was higher in the GDA-R group than in the GDA-L (95.18 vs.76.88%, p < 0,001). We observed an increase in transfusion rate in GDA-R (p < 0.05), which did not result in a higher rate of exploration (p = 0.38). CCI® patient morbidity was also significantly lower in the GDA-R group (s < 0.05). CONCLUSIONS: This study identified improved intraoperative duodenal tissue oxygen saturation in the GDA-R group with an associated reduction in pancreatic leaks and CCI® morbidity risk. A larger prospective multicenter study comparing the two methods is warranted.

Zeolite Nanoparticles Loaded with 2-Methoxystradiol as a Novel Drug Delivery System for the Prostate Cancer Therapy.

The estrogen metabolite 2-methoxyestradiol (2ME) is a promissory anticancer drug mainly because of its pro-apoptotic properties in cancer cells. However, the therapeutic use of 2ME has been hampered due to its low solubility and bioavailability. Thus, it is necessary to find new ways of administration for 2ME. Zeolites are inorganic aluminosilicates with a porous structure and are considered good adsorbents and sieves in the pharmaceutical field. Here, mordenite-type zeolite nanoparticles were loaded with 2ME to assess its efficiency as a delivery system for prostate cancer treatment. The 2ME-loaded zeolite nanoparticles showed an irregular morphology with a mean hydrodynamic diameter of 250.9 ± 11.4 nm, polydispersity index of 0.36 ± 0.04, and a net negative surface charge of -34 ± 1.73 meV. Spectroscopy with UV-vis and Attenuated Total Reflectance Infrared Fourier-Transform was used to elucidate the interaction between the 2ME molecules and the zeolite framework showing the formation of a 2ME-zeolite conjugate in the nanocomposite. The studies of adsorption and liberation determined that zeolite nanoparticles incorporated 40% of 2ME while the liberation of 2ME reached 90% at pH 7.4 after 7 days. The 2ME-loaded zeolite nanoparticles also decreased the viability and increased the mRNA of the 2ME-target gene F-spondin, encoded by SPON1, in the human prostate cancer cell line LNCaP. Finally, the 2ME-loaded nanoparticles also decreased the viability of primary cultures from mouse prostate cancer. These results show the development of 2ME-loaded zeolite nanoparticles with physicochemical and biological properties compatible with anticancer activity on the human prostate and highlight that zeolite nanoparticles can be a good carrier system for 2ME.

Micro-vibration results in vitro-derived bovine blastocysts with greater cryotolerance, epigenetic abnormalities, and a massive transcriptional change.

Much effort has been employed to improve the quality of embryos obtained by in vitro production (IVP) given the relevance of this technology to current livestock systems. In this context, dynamic IVP systems have proved beneficial to the embryo once they mimic fluid flows and mechanical forces resulting from the movement of ciliated cells and muscle contraction in the reproductive tract. In the present study, we sought to confirm these initial findings as well as assess potential molecular consequences to the embryo by applying micro-vibration (45 Hz for 5 s once per 60 min) during both oocyte maturation and embryo culture in cattle. As a result, micro-vibration led to lower incidence of apoptosis in blastocysts following vitrification-thawing. Further analyses revealed epigenetic and transcriptional changes in blastocysts derived from the micro-vibration treatment, with a total of 502 differentially expressed genes. Enrichment analyses linked differentially expressed genes to 'Oxidative phosphorylation', 'Cytokine-cytokine receptor interaction', and 'Signaling pathways regulating pluripotency of stem cells'. Yet, a meta-analysis indicated that the transcriptional changes induced by micro-vibration were not toward that of in vivo-derived embryos. In conclusion, micro-vibration increases the cryoresistance of bovine embryos, but caution should be taken given the unclear consequences of epigenetic and transcriptional abnormalities induced by the treatment.

MicroRNA Profiling Using a PCR-Based Method.

MicroRNAs (miRNAs) are small non-coding RNA molecules involved in the post-transcriptional regulation of specific mRNA targets, thus possibly controlling many biological processes. The miRNA profiling analysis can contribute to understanding several signaling pathways, as biomarkers for molecular diagnostic, as well as potential to be used as therapeutic targets. The miRNAs expression can be analyzed by quantitative reverse transcription PCR (RT-qPCR), microarrays, and RNA sequencing. The RT-qPCR method is sensitive and specific and has a lower cost when compared to other techniques as microarrays and RNA sequencing. Therefore, the protocol presented in this chapter describes step by step all the details to perform miRNA analysis using primer-based RT-qPCR.

Essential amino acid recommendations for Isa Brown layers during peak and post peak production.

The present study was designed to re-evaluate the ideal amino acid ratios of total sulphur amino acids (TSAA), Thr, Val, Ile, Trp, and Arg relative to Lys during peak and post-peak production phases in laying hens by using seven independent amino acid assays in similar experimental setting. A total of 348 twenty wk old Isa Brown laying hens were allocated to individual battery cages. Each dietary treatment included 6 replicates with 2 single cages (2 birds) as one replicate. All diets were formulated based on maize, soybean meal, and canola meal to have identical crude protein (120 g/kg) concentrations and energy density (11.9 MJ/kg) but with 5 levels of dietary concentrations of tested amino acids. Hens were offered experimental diets from 27 to 33 wk of age in experiment 1 (Exp. 1) and from 42 to 48 wk of age in experiment 2 (Exp. 2). Daily egg production and weekly egg weights were recorded, and feed intakes were calculated for each experimental period to determine egg production rate, egg mass, and feed conversion ratio (FCR). Linear and quadratic broken line models were used to estimate amino acid requirements on egg production rate, egg mass and FCR. Overall, quadratic broken line models estimated higher amino acid requirements for egg mass, egg production rate and FCR than linear broken line models by 23, 25, and 20%, respectively. The predicted daily Lys intake recommendation was 720 mg/bird/day with linear broken line model and 897 mg/bird/day with quadratic broken line model and the recommended ideal amino acid ratios relative to Lys are 85 for TSAA, 69 for Thr, 83 for Val, 87 for Ile, 22 for Trp, and 82 for Arg based on linear broken line model and 87 for TSAA, 67 for Thr, 83 for Val, 86 for Ile, 22 for Trp, and 78 for Arg based on quadratic broken line model estimations.

Obesity, inflammation, and cancer in dogs: Review and perspectives.

Obesity is the most common nutritional disease in dogs, and its prevalence has increased in recent decades. Several countries have demonstrated a prevalence of obesity in dogs similar to that observed in humans. Chronic low-grade inflammation is a prominent basis used to explain how obesity results in numerous negative health consequences. This is well known and understood, and recent studies have pointed to the association between obesity and predisposition to specific types of cancers and their complications. Such elucidations are important because, like obesity, the prevalence of cancer in dogs has increased in recent decades, establishing cancer as a significant cause of death for these animals. In the same way, intensive advances in technology in the field of human and veterinary medicine (which even proposes the use of animal models) have optimized existing therapeutic methods, led to the development of innovative treatments, and shortened the time to diagnosis of cancer. Despite the great challenges, this review aims to highlight the evidence obtained to date on the association between obesity, inflammation, and cancer in dogs, and the possible pathophysiological mechanisms that link obesity and carcinogenesis. The potential to control cancer in animals using existing knowledge is also presented.

Dietary crude protein concentrations, feed grains, and whey protein interactively influence apparent digestibility coefficients of amino acids, protein, starch, and performance of broiler chickens.

The present study was designed to investigate the impacts of dietary crude protein (CP) concentrations (220 and 180 g/kg) in either maize- or wheat-based diets, without or with 25 g/kg inclusions of whey powder (WP) concentrate on performance parameters and apparent amino acid digestibility coefficients in broiler chickens. The maize and wheat used in this study had CP levels of 84 and 119 g/kg, respectively. The 2 × 2 × 2 factorial array of 8 dietary treatments was offered to a total of 336 off-sex, male Ross 308 chicks from 7 to 35 d post-hatch with 7 replicate cages (6 birds per cage) per treatment. A treatment interaction (P = 0.016) between dietary CP and feed grains was detected for weight gains, where birds offered 180 g/kg maize-based diets displayed a weight gain advantage of 6.74% (2,628 vs. 2,462 g/bird) compared to their wheat-based counterparts. An interaction (P = 0.022) between feed grains and whey protein was observed for FCR as the addition of WP to maize-based diets improved FCR by 3.45% (1.314 vs. 1.361), but compromised FCR in wheat-based diets by 2.98% (1.415 vs. 1.374). A treatment interaction (P = 0.038) between dietary CP and feed grains was recorded for relative abdominal fat-pad weights weight gains as birds offered 180 g/kg CP maize-based diets had 43.4% (11.17 vs. 7.79 g/kg) heavier fat-pads than their wheat-based counterparts. Following the reduction in dietary-CP, apparent amino acid digestibility coefficients were depressed to greater extents in wheat-based diets. However, significant interactions between CP and feed grains were found in 14 of the 16 amino acids assessed and significant interactions between CP and WP were observed for 15 amino acids. Maize was the more suitable feed grain in terms of weight gain and FCR in 180 g/kg CP diets despite causing greater fat deposition. The inclusion of WP in reduced-CP diets did not enhance bird performance. Data generated indicate concentrations of microbial amino acids in distal ileal digesta were depressing apparent amino acid digestibility coefficients, which was more evident in wheat-based diets. Higher gut viscosities in birds offered wheat-based diets may have facilitated the proliferation of microbiota along the small intestine.

Cellular responses and microRNA profiling in bovine spermatozoa under heat shock.

In brief: Elevated temperatures disturbed sperm physiology. Bovine sperm cells exposed to heat shock led to diminished mitochondrial activity, fertilizing ability, increased oxidative stress and caspase activity concomitant with a delay in embryonic developmental kinetics and modulation of sperm-borne microRNAsmiRNAs. Abstract: Sperm function is susceptible to adverse environmental conditions. It has been demonstrated that in vivo and in vitro exposure of bovine sperm to elevated temperature reduces sperm motility and fertilizing potential. However, the cascade of functional, cellular, and molecular events triggered by elevated temperature in the mature sperm cell remains not fully understood. Therefore, the aim of this study was to determine the effect of heat shock on mature sperm cells. Frozen-thawed Holstein sperm were evaluated immediately after Percoll purification (0 h non-incubation control) or after incubation at 35, 38.5, and 41°C for 4 h. Heat shock reduced sperm motility after 3-4 h at 41°C while mitochondrial activity was reduced by 38.5 and 41°C when compared to the control. Heat shock also increased sperm reactive oxygen species production and caspase activity. Heat-shocked sperm had lower fertilizing ability, which led to diminished cleavage and blastocyst rates. Preimplantation embryo developmental kinetics was also slowed and reduced by sperm heat shock. The microRNA (miR) profiling identified >300 miRs in bovine sperm. Among these, three and seven miRs were exclusively identified in sperm cells exposed to 35 and 41°C, respectively. Moreover, miR-181d was enriched in sperm cells exposed to higher temperatures. Hence, elevated temperature altered the physiology of mature sperm cells by perturbing cellular processes and the miR profile, which collectively led to lower fertilizing ability and preimplantation development.

Vitamin D: a potentially important secosteroid for coping with COVID-19.

COVID-19 is a disease that has caused a high number of deaths in the world, and despite being controlled, it requires attention and the search for new quick and economical therapeutic strategies. In this sense, vitamin D stands out, an immunomodulator that has shown beneficial effects in decreasing the risk and severity of acute respiratory tract infections, including COVID-19. Therefore, this review presents a number of experimental, observational and clinical studies on the importance of vitamin D against viral infections with an emphasis on COVID-19, highlighting the relationship between vitamin D, Renin-Angiotensin System and cytokine storms with decreased inflammatory lesions in patients with COVID-19. In addition, aspects of pathophysiology, metabolism, risk factors, sources and recommendations of vitamin D are described. We conclude that vitamin D plays a protective role against inflammatory lesions and can decrease the risk of infections and the severity of COVID-19. Therefore, it is essential to maintain adequate levels of vitamin D to avoid complications related to its deficiency.

Characterization of histone lysine ß-hydroxybutyrylation in bovine tissues, cells, and cumulus-oocyte complexes.

Besides their canonical roles as energy sources, short-chain fatty acids act as metabolic regulators of gene expression through histone posttranslational modifications. Ketone body ß-hydroxybutyrate (BHB) causes a novel epigenetic modification, histone lysine ß-hydroxybutyrylation (Kbhb), which is associated with genes upregulated in starvation-responsive metabolic pathways. Dairy cows increase BHB in early lactation, and the effects of this increase on cellular epigenomes are unknown. We searched for and identified that Kbhb is present in bovine tissues in vivo and confirmed that this epigenetic mark is responsive to BHB in bovine and human fibroblasts cultured in vitro in a dose-dependent manner. Maturation of cumulus-oocyte complexes with high concentrations of BHB did not affect the competence to complete meiotic maturation or to develop until the blastocyst stage. BHB treatment strongly induced H3K9bhb in cumulus cells, but faintly in oocytes. RNA-seq analysis in cumulus cells indicated that BHB treatment altered the expression of 345 genes. The downregulated genes were mainly involved in glycolysis and ribosome assembly pathways, while the upregulated genes were involved in mitochondrial metabolism and oocyte development. The genes and pathways altered by BHB will provide entry points to carry out functional experiments aiming to mitigate metabolic disorders and improve fertility in cattle.

A multivariate Box-Behnken assessment of elevated branched-chain amino acid concentrations in reduced crude protein diets offered to male broiler chickens.

In a Box-Behnken assessment of elevated branched-chain amino acids (BCAA), 13 nutritionally equivalent maize-based diets were offered to a total of 390 off-sex male Ross 308 broiler chickens from 7 to 28 days post-hatch. The BCAA concentrations investigated in reduced-crude protein diets were 12.5, 15.5, 18.3 g/kg leucine (125, 155, 183); 8.9, 10.2, 12.5 g/kg valine (89, 102, 125) and 7.2, 8.9, 10.8 g/kg isoleucine (72, 89, 109), where their relativity to 11.0 g/kg digestible lysine are shown in parentheses. Determined parameters included growth performance, relative abdominal fat-pad weights, nutrient utilisation, apparent digestibility coefficients, disappearance rates of 16 amino acids and free amino acid systemic plasma concentrations. Increasing dietary leucine linearly depressed weight gain and quadratically influenced FCR where the estimated minimum FCR of 1.418 was with 14.99 g/kg leucine. Polynomial regression analysis and surface response curves of determined parameters were generated for significant (P < 0.05) BCAA variables, based on lack of fit (P > 0.005). Quadratic and cross-product responses were observed for weight gain, FCR, AME, AMEn, N retention and apparent digestibility of 13 amino acids. Relative fat-pad weights declined linearly with increasing isoleucine and valine. The lowest N retention was estimated at a combination of 15.25 and 10.50 g/kg leucine and valine respectively whilst the highest mean digestibility coefficient (0.793) of amino acids was estimated at a combination of 15.74 and 10.47 g/kg of leucine and valine respectively. The remaining parameter minima or maxima responses were not able to be determined since they were outside the extreme BCAA treatment levels. Increasing dietary BCAA significantly increased apparent ileal digestibilities and disappearance rates of BCAA. Systemic plasma concentrations of valine increased (P < 0.001) with increasing dietary valine but leucine was not influenced (P > 0.25). Systemic plasma concentration of isoleucine was maximised (P < 0.001) only when accompanied by elevated dietary leucine. Also, dietary treatments influenced (P < 0.05) apparent disappearance rates of all the essential amino acids analysed, with the exception of methionine. Whilst overall growth performance was not disadvantaged (P > 0.10) by elevated BCAA levels, compared with 2019 Ross 308 performance objectives, polynomial regression analysis suggested both interaction and antagonism between BCAA.

Enhanced removal of mercury and lead by a novel and efficient surface-functionalized imogolite with nanoscale zero-valent iron material.

A novel hybrid nanomaterial, nanoscale zero-valent iron (nZVI)-grafted imogolite nanotubes (Imo), was synthesized via a fast and straightforward chemical procedure. The as-obtained nanomaterial (Imo-nZVI) was characterized using transmission electron microscopy (TEM), electrophoretic mobility (EM), and vibrating sample magnetometry (VSM). The prepared Imo-nZVI was superparamagnetic at room temperature and could be easily separated by an external magnetic field. Sorption batch experiments were performed for single- and multicomponent systems and demonstrated that Hg2+ and Pb2+ could be quantitatively adsorbed at pH 3.0. For multicomponent systems, maximum adsorption capacities of 61.6 mg·g-1 and 76.9 mg·g-1 were obtained for Hg2+ and Pb2+ respectively. It was observed that the functional groups in Imo-nZVI interact preferentially with analytes according to the Misono softness parameter. The higher performance of Imo-nZVI compared with Imo and nZVI is related to the increased number of adsorption sites in the functionalized nanomaterial. The sorption equilibrium data obeyed the Langmuir model, while kinetic studies demonstrated that the sorption processes of Hg2+ and Pb2+ followed the pseudo-second-order model. This study suggests that the Imo-nZVI composite can be used as a promising sorbent to provide a simple and fast separation method to remove Hg and Pb ions from contaminated water.

Polyacetylene Glycosides: Isolation, Biological Activities and Synthesis.

Polyacetylene glycosides (PAGs) constitute a relatively small class of secondary metabolites characterized by the presence of a sugar unit anomerically connected to a polyacetylene. These compounds are found in fungi, seaweed, and more often in plants. PAGs exhibit a wide range of biological and pharmacological activities and, as a result, the literature of these compounds has grown exponentially in recent years.

Metagenomic analysis of microbial communities across a transect from low to highly hydrocarbon-contaminated soils in King George Island, Maritime Antarctica.

Soil samples from a transect from low to highly hydrocarbon-contaminated soils were collected around the Brazilian Antarctic Station Comandante Ferraz (EACF), located at King George Island, Antarctica. Quantitative PCR (qPCR) analysis of bacterial 16S rRNA genes, 16S rRNA gene (iTag), and shotgun metagenomic sequencing were used to characterize microbial community structure and the potential for petroleum degradation by indigenous microbes. Hydrocarbon contamination did not affect bacterial abundance in EACF soils (bacterial 16S rRNA gene qPCR). However, analysis of 16S rRNA gene sequences revealed a successive change in the microbial community along the pollution gradient. Microbial richness and diversity decreased with the increase of hydrocarbon concentration in EACF soils. The abundance of Cytophaga, Methyloversatilis, Polaromonas, and Williamsia was positively correlated (p-value = <.05) with the concentration of total petroleum hydrocarbons (TPH) and/or polycyclic aromatic hydrocarbons (PAH). Annotation of metagenomic data revealed that the most abundant hydrocarbon degradation pathway in EACF soils was related to alkyl derivative-PAH degradation (mainly methylnaphthalenes) via the CYP450 enzyme family. The abundance of genes related to nitrogen fixation increased in EACF soils as the concentration of hydrocarbons increased. The results obtained here are valuable for the future of bioremediation of petroleum hydrocarbon-contaminated soils in polar environments.

Characterization and profiling analysis of bovine oviduct and uterine extracellular vesicles and their miRNA cargo through the estrous cycle.

Extracellular vesicles (EVs) found in various biological fluids and particularly in reproductive fluids, have gained considerable attention for their possible role in cell- to- cell communication. Among, the different bioactive molecules cargos of EVs, MicroRNAs (miRNAs) are emerging as promising diagnostic biomarkers with high clinical potential. Aiming to understand the roles of EVs in bovine reproductive tract, we intended to characterize and profile the EVs of oviduct and uterine fluids (OF-EVs, UF-EVs) and their miRNA across the estrous cycle. Nanoparticle tracking analysis and transmission electron microscopy confirmed the existence of small EV population in OF and UF at all stages, (size between 30 and 200 nm; concentration: 3.4 × 1010  EVs/ml and 6.0 × 1010  EVs/ml for OF and UF, respectively, regardless of stage). The identification of EV markers (CD9, HSP70, and ALIX proteins) was confirmed by western blot. The miRNA analysis revealed the abundance of 310 and 351 miRNAs in OF-EVs and UF-EVs, respectively. Nine miRNAs were differentially abundant in OF-EVs between stages of the cycle, eight of them displayed a progressive increase from S1 to S4 (p < .05). In UF-EVs, a total of 14 miRNAs were differentially abundant between stages. Greater differences were observed between stage 1 (S1) and stage 3 (S3), with 11 miRNAs enriched in S3 compared to S1. Functional enrichment analysis revealed the involvement of these miRNAs in relevant pathways such as cell signaling, intercellular junctions, and reproductive functions that may be implicated in oviduct and uterus modulation across the cycle, but also in their preparation for embryo/conceptus presence and development.