Provision of choline chloride to the bovine preimplantation embryo alters postnatal body size and DNA methylation†.

Choline is a vital micronutrient. In this study, we aimed to confirm, and expand on previous findings, how choline impacts embryos from the first 7 days of development to affect postnatal phenotype. Bos indicus embryos were cultured in a choline-free medium (termed vehicle) or medium supplemented with 1.8 mM choline. Blastocyst-stage embryos were transferred into crossbred recipients. Once born, calves were evaluated at birth, 94 days, 178 days, and at weaning (average age = 239 days). Following weaning, all calves were enrolled into a feed efficiency trial before being separated by sex, with males being slaughtered at ~580 days of age. Results confirm that exposure of 1.8 mM choline chloride during the first 7 days of development alters postnatal characteristics of the resultant calves. Calves of both sexes from choline-treated embryos were consistently heavier through weaning and males had heavier testes at 3 months of age. There were sex-dependent alterations in DNA methylation in whole blood caused by choline treatment. After weaning, feed efficiency was affected by an interaction with sex, with choline calves being more efficient for females and less efficient for males. Calves from choline-treated embryos were heavier, or tended to be heavier, than calves from vehicle embryos at all observations after weaning. Carcass weight was heavier for choline calves and the cross-sectional area of the longissimus thoracis muscle was increased by choline.

Long-term effects of widespread pharmaceutical pollution on trade-offs between behavioural, life-history and reproductive traits in fish.

In our rapidly changing world, understanding how species respond to shifting conditions is of paramount importance. Pharmaceutical pollutants are widespread in aquatic ecosystems globally, yet their impacts on animal behaviour, life-history and reproductive allocation remain poorly understood, especially in the context of intraspecific variation in ecologically important traits that facilitate species' adaptive capacities. We test whether a widespread pharmaceutical pollutant, fluoxetine (Prozac), disrupts the trade-off between individual-level (co)variation in behavioural, life-history and reproductive traits of freshwater fish. We exposed the progeny of wild-caught guppies (Poecilia reticulata) to three field-relevant levels of fluoxetine (mean measured concentrations: 0, 31.5 and 316 ng/L) for 5 years, across multiple generations. We used 12 independent laboratory populations and repeatedly quantified activity and risk-taking behaviour of male guppies, capturing both mean behaviours and variation within and between individuals across exposure treatments. We also measured key life-history traits (body condition, coloration and gonopodium size) and assessed post-copulatory sperm traits (sperm vitality, number and velocity) that are known to be under strong sexual selection in polyandrous species. Intraspecific (co)variation of these traits was analysed using a comprehensive, multivariate statistical approach. Fluoxetine had a dose-specific (mean) effect on the life-history and sperm trait of guppies: low pollutant exposure altered male body condition and increased gonopodium size, but reduced sperm velocity. At the individual level, fluoxetine reduced the behavioural plasticity of guppies by eroding their within-individual variation in both activity and risk-taking behaviour. Fluoxetine also altered between-individual correlations in pace-of-life syndrome traits: it triggered the emergence of correlations between behavioural and life-history traits (e.g. activity and body condition) and between life-history and sperm traits (e.g. gonopodium size and sperm vitality), but collapsed other between-individual correlations (e.g. activity and gonopodium size). Our results reveal that chronic exposure to global pollutants can affect phenotypic traits at both population and individual levels, and even alter individual-level correlations among such traits in a dose-specific manner. We discuss the need to integrate individual-level analyses and test behaviour in association with life-history and reproductive traits to fully understand how animals respond to human-induced environmental change.

Immunohistochemical examination of the utero-placental interface of cows on days 21, 31, 40, and 67 of gestation.

What we understand about early stages of placentation in cattle is based on an elegant series of electron microscopic images that provide exquisite detail, but limited appreciation for the microanatomy across the utero-placental interface. In order to achieve a global perspective on the histology of bovine placentation during critical early stages of gestation, i.e., days 21, 31, 40, and 67, we performed immunohistochemistry to detect cell-specific expression of pregnancy-associated glycoprotein (PAG), cytokeratin, epithelial (E)-cadherin, and serine hydroxymethyltransferase 2 (SHMT2) at the intact utero-placental interface. Key findings from the immunohistochemical analyses are that there are: (1) PAG-positive cells with a single nucleus within the uterine luminal epithelial (LE) cells; (2) PAG-positive cells with two nuclei in the LE; (3) PAG-positive syncytial cells with more than three nuclei in the LE; (4) LE cells that are dissociated from one another and dissociated from the basement membrane in regions of syncytialization within the LE layer; (5) replacement of the mononuclear LE with a multi-layer thick population of PAG-positive cells invading into the uterine stroma of caruncles, but not into the stroma of intercaruncular endometrium; and (6) PAG-, E-cadherin- and SHMT2-positive mononuclear cells at the leading edge of developing cotyledonary villi that eventually represent the majority of the epithelial surface separating caruncular stroma from cotyledonary stroma. Finally, the utero-placental interface of ruminants is not always uniform across a single cross-section of a site of placentation which allows different conclusions to be made depending on the part of the utero-placental interface being examined.

Actions of DKK1 on the preimplantation bovine embryo to affect pregnancy establishment, placental function, and postnatal phenotype†.

One mechanism by which the maternal environment regulates the early embryo is by secretion of cell-signaling molecules. One of these is dickkopf WNT signaling pathway inhibitor 1. Objectives were to (A) resolve discrepancies in the literature regarding effects of dickkopf WNT signaling pathway inhibitor 1 in the bovine embryo on development of trophectoderm and competence to establish pregnancy after embryo transfer and (B) determine whether there are long-term consequences of dickkopf WNT signaling pathway inhibitor 1 on placental function and postnatal phenotype. Embryos produced in vitro were cultured with vehicle or 100 ng/mL recombinant human dickkopf WNT signaling pathway inhibitor 1 from Days 5 to 7.5 of development (i.e., the morula and blastocyst stages of development). dickkopf WNT signaling pathway inhibitor 1 increased the number of cells positive for the trophectoderm marker CDX2 at Day 7.5 of development while having no effect on number of cells positive for the inner cell mass marker SOX2. There was no effect of dickkopf WNT signaling pathway inhibitor 1 on pregnancy or calving rate after transfer of blastocysts produced with Y-sorted semen to either lactating dairy cows or suckling beef cows. Treatment with dickkopf WNT signaling pathway inhibitor 1 at the morula-to-blastocyst stages programmed placental function, as measured by an effect of dickkopf WNT signaling pathway inhibitor 1 on plasma concentrations of pregnancy associated glycoproteins and placental lactogen at Day 160 of gestation (although not on other days examined). dickkopf WNT signaling pathway inhibitor 1 treatment also resulted in calves that were heavier at birth as compared to calves derived from control embryos. After birth, dickkopf WNT signaling pathway inhibitor 1 calves grew slower than controls. Results confirm that dickkopf WNT signaling pathway inhibitor 1 alters the developmental program of the bovine embryo to affect both prenatal and postnatal phenotypes.

Cold Atmospheric Plasma Jet as a Possible Adjuvant Therapy for Periodontal Disease.

Due to the limitations of traditional periodontal therapies, and reported cold atmospheric plasma anti-inflammatory/antimicrobial activities, plasma could be an adjuvant therapy to periodontitis. Porphyromonas gingivalis was grown in blood agar. Standardized suspensions were plated on blood agar and plasma-treated for planktonic growth. For biofilm, dual-species Streptococcus gordonii + P. gingivalis biofilm grew for 48 h and then was plasma-treated. XTT assay and CFU counting were performed. Cytotoxicity was accessed immediately or after 24 h. Plasma was applied for 1, 3, 5 or 7 min. In vivo: Thirty C57BI/6 mice were subject to experimental periodontitis for 11 days. Immediately after ligature removal, animals were plasma-treated for 5 min once-Group P1 (n = 10); twice (Day 11 and 13)-Group P2 (n = 10); or not treated-Group S (n = 10). Mice were euthanized on day 15. Histological and microtomography analyses were performed. Significance level was 5%. Halo diameter increased proportionally to time of exposure contrary to CFU/mL counting. Mean/SD of fibroblasts viability did not vary among the groups. Plasma was able to inhibit P. gingivalis in planktonic culture and biofilm in a cell-safe manner. Moreover, plasma treatment in vivo, for 5 min, tends to improve periodontal tissue recovery, proportionally to the number of plasma applications.

Systemic and local effects of radiotherapy: an experimental study on implants placed in rats.

OBJECTIVES: Evaluate the modulating effect of ionizing radiation, blood cytokine levels, and bone remodeling of the interface around the implant to understand the radiation mechanisms which can impair the implants receptor site. MATERIAL AND METHODS: Sixty rats were submitted to grade V titanium implants in the femurs and were divided into the following groups: no-irradiation (N-Ir): control group with implant only; early-irradiation (E-Ir): implant + irradiation after 24 h; late-irradiation (L-Ir): implant + irradiation after 4 weeks; and previous-irradiation (P-Ir): irradiation + implant after 4 weeks. The animals in the E-Ir, L-Ir, and P-Ir groups were irradiated in two fractional stages of 15 Gy. At 3 days, 2 weeks, and 7 weeks after the final procedure, five animals were randomly euthanized per group. Serum levels of TNF-ɑ, IL-1ß, TGF-ß, IL-6, M-CSF, and IL-10 were measured from blood collected prior to euthanasia using the ELISA test. The pieces containing the implants were subjected to immunohistochemical labeling using the tartrate acid resistant to phosphatase, osteocalcin, and caspase-3 markers and mCT. The ANOVA test was used for statistical analysis, and the Tukey multiple comparison test (p < 0.05) was applied. RESULTS: The results indicated that ionizing radiation modifies the production of pro- and anti-inflammatory serum cytokines, the expression of proteins involved in bone remodeling and cellular apoptosis, as well as changes in bone formation. CONCLUSIONS: The results suggests that a longer period between radiotherapy and implant placement surgery when irradiation occurs prior to implant installation would allow the recovery and renewal of bone cells and avoid future failures in osseointegration. CLINICAL RELEVANCE: The search for modifications caused by ionizing irradiation in bone tissue can indicate the ideal period for implant placement without affecting the osseointegration process.

A novel strategy for resynchronization of ovulation in Nelore cows using injectable progesterone (P4) and P4 releasing devices to perform two timed inseminations within 22 days.

We aimed to evaluate the reproductive performance of Nelore lactating cows submitted to a resynchronization 12 days after timed artificial insemination (TAI) with or without a long-acting progesterone (P4-LA) treatment. Nelore cows were submitted to a P4/oestradiol-based TAI protocol (D0 = insemination). On D12, cows in the control group (n = 184) received a new P4 intravaginal device (0.96 g), whereas cows in the P4-LA group (n = 192) received the P4 device and 75 mg P4-LA. Cows identified as non-pregnant (n = 120) by regression of corpus luteum using colour Doppler ultrasonography on D20 had the P4 device removed and received 500ug of sodium cloprostenol, 1 mg of oestradiol cypionate and 300 IU of eCG and were re-inseminated on D22. There was no difference (p > 0.10) in the pregnancy rate at D20, D30 and D60 after first TAI between the control (69%, 59.7% and 57%, respectively) and P4-LA (67%, 55.7%, and 55.2%, respectively) groups. Pregnancy losses were similar between both groups (p > 0.1). For cows submitted to the second TAI, the pre-ovulatory follicle size did not differ (p > 0.1), but the oestrous detection and pregnancy rates were greater (p < 0.05) in the P4-LA group (92.2% [59/64] and 60.9% [39/64], respectively) than in controls (75% [42/56] and 44.6% [25/56]). The cumulative pregnancy rate after two TAIs did not differ (p > 0.1) between control (73.3% [135/184]) and P4-LA (76% [146/192]) groups. The use of P4-LA at 12 days after TAI potentially increases the pregnancy rates for a new early resynchronization strategy associated with the Doppler imaging for pregnancy diagnosis and results in an alternative to perform two TAIs in 22 days in beef cows.

Effects of the GaAlAs diode laser (780 nm) on the periodontal tissues during orthodontic tooth movement in diabetes rats: histomorphological and immunohistochemical analysis.

The purposes of the present study are to assess the effects of the GaAlAs diode laser on the periodontal tissues and to investigate its action on the alveolar bone remodeling process during orthodontic tooth movement in normoglycemic and diabetic rats. Sixty adult male Wistar rats were divided into four groups of 15 rats: normoglycemic (N), diabetic (D), laser-normoglycemic (LN), and laser-diabetic (LD) rats. Diabetes mellitus was induced by a single intravenous injection of 40 mg/kg monohydrated alloxan. The orthodontically moved tooth underwent a force magnitude of 20 cN. The laser irradiation with a continuous emission of a 780-nm wavelength, an output power of 20 mW, and a fiber probe with a spot size of 0.04 cm in diameter and an area of 0.00126 cm2 were used. Moreover, an energy density of 640 J/cm2 was applied in an exposition time of 40 s. Histomorphological and immunohistochemical analysis was performed. The photobiomodulation (PBM) strongly stimulated the periodontal tissue response, establishing mainly the balance between the bone formation and resorption. Intense inflammatory cell infiltration and extensive loss of bone tissue were mainly found in the D group from 14 days. The number of osteopontin-positive osteocytes was significantly greater in the LN group, followed by the LD, especially at 7 and 14 days, whereas osteoprotegerin-positive osteoblasts were significantly higher in the LN and LD groups than in the N and D groups, respectively, in all periods. The PBM strongly stimulated the alveolar bone remodeling and favored the continuous reorganization of the soft periodontal tissues, leading to the maintenance and integrity of the periodontal microstructure under orthodontic force, especially in uncontrolled diabetic rats.

Modulation of adenosine signaling prevents scopolamine-induced cognitive impairment in zebrafish.

Adenosine, a purine ribonucleoside, exhibits neuromodulatory and neuroprotective effects in the brain and is involved in memory formation and cognitive function. Adenosine signaling is mediated by adenosine receptors (A1, A2A, A2B, and A3); in turn, nucleotide and nucleoside-metabolizing enzymes and adenosine transporters regulate its levels. Scopolamine, a muscarinic cholinergic receptor antagonist, has profound amnesic effects in a variety of learning paradigms and has been used to induce cognitive deficits in animal models. This study investigated the effects of acute exposure to caffeine (a non-selective antagonist of adenosine receptors A1 and A2A), ZM 241385 (adenosine receptor A2A antagonist), DPCPX (adenosine receptor A1 antagonist), dipyridamole (inhibitor of nucleoside transporters) and EHNA (inhibitor of adenosine deaminase) in a model of pharmacological cognitive impairment induced by scopolamine in adult zebrafish. Caffeine, ZM 241385, DPCPX, dipyridamole, and EHNA were acutely administered independently via i.p. in zebrafish, followed by exposure to scopolamine dissolved in tank water (200µM). These compounds prevented the scopolamine-induced amnesia without impacting locomotor activity or social interaction. Together, these data support the hypothesis that adenosine signaling may modulate memory processing, suggesting that these compounds present a potential preventive strategy against cognitive impairment.

Conventional and whitening toothpastes: cytotoxicity, genotoxicity and effect on the enamel surface.

PURPOSE: To evaluate the cytotoxicity and genotoxicity of whitening and common toothpastes, and the surface roughness of tooth enamel submitted to brushing with both toothpastes. METHODS: Samples of whitening toothpastes [Colgate Whitening (CW) and Oral-B Whitening (OBW)] and regular (non-whitening) toothpastes (Colgate and Oral-B) were extracted in culture medium. Gingival human fibroblasts (FMM-1) were placed in contact with different dilutions of culture media that had been previously exposed to such materials, and the cytotoxicity was evaluated using the MTT assay. The genotoxicity was assessed by the micronucleus formation assay in Chinese hamster fibroblasts (V79). The cell survival rate and micronuclei number were assessed before and after exposure to the toothpaste extracts. For the surface roughness evaluation, 20 bovine tooth specimens, divided into four groups according to toothpastes, were submitted to 10,000 brushing cycles. The results were analyzed using the Mann-Whitney U and two-way ANOVA tests (P < 0.05). RESULTS: MTT assay showed that Colgate was significantly less cytotoxic than CW, Oral-B and OBW at all dilutions (P < 0.01). CW was the most cytotoxic toothpaste (P < 0.01). The whitening toothpastes showed the highest numbers of micronuclei compared to the untreated control (UC) (P < 0.01). Colgate and Oral-B toothpastes were not genotoxic compared to UC (P = 0.326). The OBW toothpaste was statistically significantly abrasive to the enamel surface (P < 0.01). The whitening toothpastes and Oral-B were cytotoxic to the cells. The whitening toothpastes were more genotoxic to cells in vitro than the common toothpastes, and genotoxicity was more pronounced in the OBW toothpaste.

Vitamin D and follicular recruitment in the in vitro fertilization cycle.

OBJECTIVE: Vitamin D (VD) is a fat-soluble steroid hormone, synthesized by the skin, most known for its role in bone mineral balance. Vitamin D receptors (VDR) are also found in the female reproductive system, but their role remains unclear. The objective of this study was to analyze the relationship between serum vitamin D levels and the number of oocytes retrieved after ovarian stimulation. METHODS: This is a retrospective study involving 267 patients undergoing in vitro fertilization (IVF) carried out in the Fertipraxis clinic, a private practice facility. The patients were initially divided into two groups according to their VD levels. Group 1 included 152 patients with VD levels < 30 ng/mL and group 2 had 115 patients with VD levels > 30 ng/mL. They were further analyzed and separated considering their age, anthropometric data, ovarian reserve, amount of gonadotropin used, and follicles obtained until trigger day. RESULTS: In our analysis, there were no difference in the number of follicles and oocytes retrieved, nor in the number of mature oocytes obtained from patients with both vitamin D deficiency and sufficiency. CONCLUSIONS: The results of our study show no difference among number of follicles, oocytes retrieved and mature oocytes obtained after ovarian stimulation according to their vitamin D serum levels. Further higher-quality studies are needed to evaluate the possible roles of serum vitamin D levels in other stages of human fertilization process.

Vitamin D levels in couples undergoing in vitro fertilization treatment: lack of association with embryo quality or pregnancy rates.

OBJECTIVE: To investigate the potential impact of vitamin D (VD) serum levels on couples going through in vitro fertilization treatment in terms of embryo quality and pregnancy rates. DESIGN: A retrospective cohort study. SETTING: A private human reproduction center. PATIENT(S): A total of 267 couples underwent intracytoplasmic sperm injections between January 2017 and March 2019. INTERVENTION(S): The couples were categorized into four groups on the basis of 25-hydroxy VD (25OHD) levels measured at the beginning of the stimulation protocol: group 1 with 25OHD levels ≥30 ng/mL for both women and men; group 2 with 25OHD levels <30 ng/mL for both; group 3 women with 25OHD levels <30 ng/mL and men with 25OHD levels ≥30 ng/mL; and group 4 with women with 25OHD level ≥30 ng/mL and men with 25OHD level <30 ng/mL. MAIN OUTCOME MEASURE(S): We consider the quantity and quality of embryos during the cleavage as well as blastocyst stages as primary outcomes. Correspondingly, the clinical pregnancy rate (CPR) was regarded as a secondary outcome. RESULT(S): Our findings revealed no significant correlations between the studied VD groups and the evaluated outcomes. This includes the quantity and quality of embryos during the cleavage and blastocyst stages, as well as the CPR. Primary analysis revealed a small but statistically significant difference in the duration of controlled ovarian stimulation between group 1 and group 2 (95% confidence interval, 0.07-3.04) and between group 1 and group 3 (95% confidence interval, 0.05-3.23). CONCLUSION(S): The present study found no correlation between the studied VD levels and the quantity as well as quality of cleavage or blastocyst stage embryos, nor did it show any impact on CPRs. Further well-designed, prospective studies are warranted to determine whether and how vitamin D affects reproductive outcomes.

Herlyn-Werner-Wunderlich syndrome: A fertility-sparing approach to a rare mullerian anomaly.

The Herlyn-Werner-Wunderlich syndrome (HWWS) is characterized by the triad of uterus didelphys, obstructed hemivagina, and renal agenesis. The typical clinical presentation involves chronic pelvic pain, dysmenorrhea, and palpable abdominal mass, related to hematocolpos/hematometra. It is a rare disease, with a challenging clinical and radiological diagnosis. Surgery is the definitive treatment. Complications such as endometriosis, infertility and chronic pelvic pain occur more frequently and severely when diagnosis and treatment are delayed. This is a case report of a twelve-year-old patient admitted to the Gynecology Department of the Federal University of Rio de Janeiro's General Hospital (HUCFF/UFRJ), in March 2021, with progressive symptoms of dysmenorrhea and abdominal distention due to palpable abdominal mass. She had a previous history of congenital solitary kidney. Magnetic Resonance Imaging (MRI) showed a double uterus with hematometra and hematocolpos on the left side, pelvic endometriosis and left renal agenesis. Conservative clinical treatment with inhibition of the hypothalamic-pituitary-ovarian (H-P-O) axis was initiated while a definitive surgical approach was being defined. In June 2022, the patient underwent left hemi-hysterectomy and salpingectomy, achieving full remission of symptoms. Given the rarity of this syndrome and its potential complications, our report aims to familiarize clinicians with it, mostly those who work with children and adolescents, so that more patients have access to early diagnosis and adequate treatment. Consequently, future fertility can be effectively preserved.

Switching from cetrorelix to dydrogesterone in an IVF cycle - a new strategy for unexpected freeze-all cycles.

Preventing a luteinizing hormone (LH) surge is a major concern in controlled ovarian stimulation (COS). Several strategies have been developed over the years, including protocols with Gonadotrophin Releasing Hormone agonists and antagonists. More recently Progestin Primmed Ovarian Stimulation (PPOS) has shown to be equally effective in pituitary suppression, with comparable clinical and laboratorial outcomes. This is the case of a 34 year old female, with a previous diagnosis of primary infertility due to tubal factor and high ovarian reserve markers. The initial plan was to perform IVF/ICSI. followed by fresh blastocyst transfer. The chosen COS strategy was to use Alfacorifolitropin 150mg (Elonva®) and Cetrorelix acetate 0,25mg (Cetrotide®) in a flexible pituitary suppression protocol. However, because of elevated risk for Ovarian Hyper-stimulation Syndrome (OHSS) detected during ultrasound and hormonal monitoring, in order to diminish financial burden and to have a more patient friendly protocol, we switched cetrorelix acetate to oral dydrogesterone. COS was successful and resulted in 24 retrieved oocytes (16 metaphase 2 oocytes) without any premature LH peak. No OHSS symptoms occurred. Our main goal with this case report is to reinforce the feasibility and efficacy of this innovative approach, especially in patients aiming for a fresh embryo transfer, who present alert sings of OHSS during the stimulation. Developing friendlier and cheaper protocols in assisted reproduction makes the treatment more accessible and affordable.

Development of ring radiation dosemeters using 3D printing.

Extremity radiation monitoring is an important tool for the assessment of occupational exposures to staff at a variety of workplaces where ionising radiation is used. This work shows the feasibility of applying 3D printing for the development of customisable ring dosemeters. The rings were developed using two types of resin, hard and flexible and has the possibility of sterilisation using different techniques. The printed ring dosemeter was associated with BeO optically stimulated dosemeters. The energy and angular response were found within ±20% in the energy range from 24 to 662 keV and from 0° to 60° angle of incidence. This contributes to the reduction of measurement uncertainty when compared with currently used thermoluminescent detectors dosemeters. The new ring dosemeter showed a satisfactory response with respect to the performance criteria of the IEC 62387 Standard, in addition to providing improved ergonomics in relation to the commercial ring dosemeter.

Reproductive microbiome and cytokine profiles associated with fertility outcomes of postpartum beef cows.

Shifts from commensal bacteria (for example, Lactobacillus in the phylum Firmicutes) within the reproductive tract have been associated with changes in local reproductive immune responses and decreased fertility in humans. The objective of this study was to characterize the microbiome and cytokine concentrations before artificial insemination (AI) in vaginal and uterine flushes from postpartum beef cows. Twenty Bos indicus-influenced beef cows (approximately 60 d postpartum and free of reproductive, health, or physical issues) were enrolled. The B. indicus prostaglandin (PG) 5-d + controlled intervaginal drug-releasing estrus synchronization protocol was initiated on day -8 of the study with timed AI on d0. Blood samples were collected on days -3, -1, and 28 via coccygeal venipuncture. Vaginal and uterine flushes were collected on days -3 and -1. Based on days 28 pregnancy status determined by transrectal ultrasonography, cows were identified as either Open (n = 13) or Pregnant (n = 7). Bacterial community analyses were conducted targeting the V4 hypervariable region of the 16S rRNA gene. Cytokine analyses were performed using the RayBiotech Quantibody Bovine Cytokine Array Q1 and MyBioSource ELISA kits per the manufacturer's instructions. Statistical analyses for bacteria relative abundance were conducted using PROC NPAR1WAY and for cytokine concentrations using PROC GLM in SAS 9.4. Uterine concentrations of interferon γ, interleukin (IL)1α, and IL21 were greater in Open than in Pregnant cows (P < 0.05). Regardless of pregnancy status, uterine IL13 increased from days -3 to -1 (9.76 vs. 39.48 ± 9.28 pg/mL, respectively; P < 0.05). Uterine relative abundance of the phylum Firmicutes decreased from days -3 to -1 in Open cows (60.4% ± 0.9% vs. 48.5% ± 3.2%; P = 0.004). In Open cows, the genus Blautia decreased in relative abundance within the uterus from days -3 to -1 (2.1% ± 0.2% vs. 0.9% ± 0.1%; P = 0.002). Uterine relative abundance of the phylum Tenericutes increased from days -3 to -1 in Pregnant cows (1.0% ± 0.1% vs. 7.6% ± 4.1%; P = 0.002). In Pregnant cows, the genus Ureaplasma tended to increase within the uterus from days -3 to -1 (0.08% ± 0.06% vs. 7.3% ± 4.1%; P = 0.054). These findings suggest a distinct difference in the reproductive microbiome and cytokine profiles before AI for resulting Open vs. Pregnant cows.

Efficiently producing cattle to feed a growing population can come with many challenges. A few challenges occur soon after a cow has given birth, and subsequent reproductive performance can be impacted. Bacteria within the reproductive tract can trigger an immune response and together play a role in affecting fertility in cows. The objectives of this experiment were to distinguish the commensal vs. harmful bacteria that reside in the reproductive tract and to characterize the immune response in beef cattle via uterine and vaginal flushes. The results demonstrated that bacteria within the reproductive tract of beef cattle changes before breeding. The current study also suggests that changes in immune response before breeding can be associated with fertility outcomes. Additional research may be worthwhile to evaluate management tactics to positively shift bacteria within the reproductive tract and reduce inflammatory immune responses to improve fertility and increase reproductive efficiency. Future research is necessary to identify the causes of bacterial shifts and how it relates to pregnancy establishment.

Sex differences in the predictability of risk-taking behavior.

Recent research has found that individuals often vary in how consistently they express their behavior over time (i.e., behavioral predictability) and suggested that these individual differences may be heritable. However, little is known about the intrinsic factors that drive variation in the predictability of behavior. Indeed, whether variation in behavioral predictability is sex-specific is not clear. This is important, as behavioral predictability has been associated with vulnerability to predation, suggesting that the predictability of behavioral traits may have key fitness implications. We investigated whether male and female eastern mosquitofish (Gambusia holbrooki) differed in the predictability of their risk-taking behavior. Specifically, over a total of 954 behavioral trials, we repeatedly measured risk-taking behavior with three commonly used assays-refuge-use, thigmotaxis, and foraging latency. We predicted that there would be consistent sex differences in both mean-level risk-taking behavior and behavioral predictability across the assays. We found that risk-taking behavior was repeatable within each assay, and that some individuals were consistently bolder than others across all three assays. There were also consistent sex differences in mean-level risk-taking behavior, with males being bolder across all three assays compared to females. In contrast, both the magnitude and direction of sex differences in behavioral predictability were assay-specific. Taken together, these results highlight that behavioral predictability may be independent from underlying mean-level behavioral traits and suggest that males and females may differentially adjust the consistency of their risk-taking behavior in response to subtle changes in environmental conditions.

Evolution of research funding for neglected tropical diseases in Brazil, 2004-2020.

Neglected tropical diseases are a global public health problem. Although Brazil is largely responsible for their occurrence in Latin America, research funding on the subject does not meet the population's health needs. The present study analyzed the evolution of research funding for neglected tropical diseases by the Ministry of Health and its partners in Brazil, from 2004 to 2020. This is a retrospective study of data from investigations registered on Health Research (Pesquisa Saúde in Portuguese), a public repository for research funded by the Ministry of Health's Department of Science and Technology. The temporal trend of funding and the influence of federal government changes on funding were analyzed using Prais-Winster generalized linear regression. From 2004 to 2020, 1,158 studies were financed (purchasing power parity (PPP$) 230.9 million), with most funding aimed at biomedical research (81.6%) and topics involving dengue, leishmaniasis and tuberculosis (60.2%). Funding was stationary (annual percent change of -5.7%; 95%CI -54.0 to 45.0) and influenced by changes to the federal government. Research funding was lacking for chikungunya, Chagas disease, schistosomiasis, malaria and taeniasis/cysticercosis, diseases with a high prevalence, burden or mortality rates in Brazil. Although the Ministry of Health had several budgetary partners, it was the main funder, with 69.8% of investments. The study revealed that research funding for neglected tropical diseases has stagnated over the years and that diseases with a high prevalence, burden and mortality rate receive little funding. These findings demonstrate the need to strengthen the health research system by providing sustainable funding for research on neglected tropical diseases that is consistent with the population's health needs.

Decreased cardiac output: an integrative review.

OBJECTIVE: to identify, in the scientific literature, the defining characteristics and contributing factors (related factors, associated conditions and populations at risk) for nursing diagnosis decreased cardiac output. METHOD: an integrative literature review, conducted between September and October 2020, with an update in March 2022, in the MEDLINE via PubMed, LILACS, SciELO, CINAHL and EMBASE databases. Using acronym PEO, studies published in the last 10 years in Portuguese, English and Spanish were included. A descriptive analysis was carried out to present the elements mapped in the literature. RESULTS: analysis of 31 articles identified different elements, highlighting 4 new related factors: hyperglycemic stress, prone position, left lateral position, sleep deprivation. Individuals with a history of cardiovascular disease and males were identified as possible populations at risk. FINAL CONSIDERATIONS: the elements for decreased cardiac output, identified in the literature, add evidence that justifies the permanence of this diagnosis in the NANDA-I classification.

Leg Ulcers in Sickle Cell Disease: A Multifactorial Analysis Highlights the Hemolytic Profile.

Sickle cell disease (SCD) is characterized by the presence of the variant S hemoglobin (HbS). The homozygous genotype (HbSS) is sickle cell anemia (SCA), while the double heterozygous of HbS and HbC (HbSC) is defined as SC hemoglobinopathy. The pathophysiology is based on chronic hemolysis, inflammation, endothelial dysfunction, and vaso-occlusion, which results in vasculopathy and serious clinical manifestations. Sickle leg ulcers (SLUs) are cutaneous lesions around the malleoli frequent in 20% of Brazilian patients with SCD. SLUs present a variable clinical and laboratory pattern modulated by several characteristics that are not fully understood. Hence, this study aimed to investigate laboratory biomarkers and genetic and clinical parameters associated with the development of SLUs. This descriptive cross-sectional study included 69 SCD patients, 52 without SLU (SLU-) and 17 with active or previous SLU history (SLU+). The results showed a higher incidence of SLU in SCA patients and there was no observed association of α-3.7 Kb thalassemia in SLU occurrence. Alterations in NO metabolism and hemolysis were associated with clinical evolution and severity of SLU, in addition to hemolysis modulating the etiology and recurrence of SLU. Our multifactorial analyses demonstrate and extend the role of hemolysis driving the pathophysiological mechanism of SLU.