Metabolic Shift in Porcine Spermatozoa during Sperm Capacitation-Induced Zinc Flux.

Mammalian spermatozoa rely on glycolysis and mitochondrial oxidative phosphorylation for energy leading up to fertilization. Sperm capacitation involves a series of well-regulated biochemical steps that are necessary to give spermatozoa the ability to fertilize the oocyte. Additionally, zinc ion (Zn2+) fluxes have recently been shown to occur during mammalian sperm capacitation. Semen from seven commercial boars was collected and analyzed using image-based flow cytometry before, after, and with the inclusion of 2 mM Zn2+ containing in vitro capacitation (IVC) media. Metabolites were extracted and analyzed via Gas Chromatography-Mass Spectrometry (GC-MS), identifying 175 metabolites, with 79 differentially abundant across treatments (p < 0.05). Non-capacitated samples showed high levels of respiration-associated metabolites including glucose, fructose, citric acid, and pyruvic acid. After 4 h IVC, these metabolites significantly decreased, while phosphate, lactic acid, and glucitol increased (p < 0.05). With zinc inclusion, we observed an increase in metabolites such as lactic acid, glucitol, glucose, fructose, myo-inositol, citric acid, and succinic acid, while saturated fatty acids including palmitic, dodecanoic, and myristic acid decreased compared to 4 h IVC, indicating regulatory shifts in metabolic pathways and fatty acid composition during capacitation. These findings underscore the importance of metabolic changes in improving artificial insemination and fertility treatments in livestock and humans.

Effect of chronic cyclic heat stress and supplemented inorganic and organic zinc source levels on grow-finish pig growth performance and estimated body composition.

Zinc (Zn) supplementation has proved to mitigate the effects of heat stress with varying effects evident with Zn source during acute heat events. However, the effects of Zn supplementation during long-term summer weather patterns have yet to be explored. Therefore, the objective of this study was to identify the effects of supplementation source and level of Zn to mitigate the negative effects of long-term, cyclic heat stress in finishing swine. Six hundred cross-bred pigs were housed under thermoneutral (TN) or cyclic heat (HS) conditions simulating summer heat with acute 3-d heat waves for a 70-d study. Thermoneutral conditions were 16.7 to 18.9 °C throughout the study. HS pigs were housed at the same temperature as TN from days 0 to 18, then 28 °C/24 °C for 12 h/12 h on days 18 to 21, followed by 30 °C/26.7 °C for 12 h/12 h on days 24 to 70, except during acute heat (32 to 33 °C/29 to 30 °C, 12 h/12 h) on days 21 to 24, 42 to 45, and 63 to 66. Treatments were arranged in a 2 × 6 factorial with main effects of environment (HS vs. TN) and dietary available Zn supplementation: (1) 50 mg/kg zinc oxide (ZnO), (2) 130 mg/kg ZnO, (3) 50 mg/kg of organic Zn (Availa Zn), (4) 50 mg/kg ZnO + 40 mg/kg organic Zn, (5) 50 mg/kg ZnO + 60 mg/kg organic Zn, and (6) 50 mg/kg ZnO + 80 mg/kg organic Zn. Pigs (5 pigs/pen) were blocked by initial body weight (72.2 kg) and randomly allotted to 1 of 12 temperature and diet treatment combinations across 10 replicates. Body weight and feed intake were determined at the beginning and end of each acute heat event. All pigs were ultrasonically scanned at the 10th rib (TR) to predict loin muscle area (LMA), backfat (BF), and percent lean. Data were analyzed by the MIXED procedure in SAS with pen as the experimental unit. At day 63, HS pigs were lighter (P < 0.05), had lower overall average daily gain (ADG; P < 0.05) and average daily feed intake (P < 0.05). A diet-by-environment interaction was observed for overall ADG (P < 0.05) with diet 5 HS pigs having a 3.9% reduction in ADG whereas diet 6 had 14.4% reduction in ADG, while under TN temperatures diet 6 had the greatest overall ADG of all treatments. Other diets were intermediate in their ADG under both HS and TN conditions. Pigs under HS had less BF at the TR (P < 0.05) and a smaller LMA (P < 0.05), and a greater calculated percent lean (P < 0.05). Our results indicate that a blend of supplemental Zn sources at 50/60 mg/kg may mitigate the reduction in growth performance due to HS. While not directly contrasted, the NRC requirement of 50 mg/kg Zn may be too low to optimize finishing pig growth performance under both TN and HS conditions.

Low colostrum intake results in potential accumulation of peroxisome lipid substrates in vaginal tissue of 3-week-old gilts.

Low colostrum intake relates to poorer health and infertility in swine. We previously connected vaginal lipid profiles at weaning to fertility of sows. We hypothesized vaginal lipidome varied with colostrum intake. Our objective was to determine whether indicators of colostrum intake, immunocrit (IM) and weight gain 24 h postnatal (PN), related to vaginal lipids at d21 PN. Gilts (n=60) were weighed and blood sampled to measure IM. On d21 PN vaginal swabs were taken and lipids measured using multiple reaction monitoring. Abundance of multiple lipids differed (P<0.05) between gilts categorized as high versus low IM and high versus low 24 h gain. The abundance of multiple lipids correlated with IM and 24 h gain. Phosphatidylcholine PC(36:3), PC(36:2), and arachidonic acid (C20:4) positively (P<0.05) correlated with IM. The ether lipid PCo(38:6) and multiple cholesteryl esters negatively (P<0.05) correlated with IM. ROC analysis indicated arachidonic acid and docosanoic acid (C22:0) may serve as excellent biomarkers that distinguish between high and low IM. Similar to gilts found to be infertile, lipid profiles of low colostrum intake animals had greater abundance of very long chain fatty acids, lipids with high levels of unsaturation, and cholesteryl esters, which are metabolized in peroxisomes indicating their potential dysfunction.

Rate of Complete Spectacle Independence with a Trifocal Intraocular Lens: A Systematic Literature Review and Meta-Analysis.

INTRODUCTION: A systematic literature review and meta-analysis was conducted to identify and obtain a precise single summary estimate on complete spectacle independence after bilateral implantation of a trifocal intraocular lens (IOL) (AcrySof PanOptix, TFNTXX/TFATXX) for patients undergoing cataract surgery. METHODS: A search was conducted in PubMed from January 2017 to September 2021. Relevant congress presentations were also searched to include data from completed studies not yet published. Search terms included the intervention (TFNTXX, TFATXX, PanOptix) and outcomes of interest (patient-reported spectacle independence rates). A Bayesian random-effects meta-analysis was conducted, providing a pooled estimate (median and its 95% credible interval) of complete spectacle independence rates among cataract surgery patients. Subgroup analyses evaluated spectacle independence after cataract surgery across different working distances (near, intermediate, far). RESULTS: Nineteen unique clinical studies were identified. Based on a meta-analysis of 13 studies (N = 513 patients), the complete spectacle independence rate after cataract surgery with TFNTXX/TFATXX IOL was 91.6% (95% credible interval 86.8-95.9%). Additionally, the spectacle independence rates at each focal point (N = 13 studies, 603 patients) were 89.6% (near), 96.3% (intermediate), and 95.9% (far). CONCLUSIONS: This meta-analysis demonstrated that at least nine out of ten patients receiving TFNTXX/TFATXX trifocal IOL during cataract surgery can expect to achieve complete spectacle independence. This study provides informative data for clinicians and patients to feel confident in the use of trifocal intraocular lenses as presbyopia-correcting IOLs that offer high rates of complete spectacle independence.

Qualitative interviews to evaluate content validity of the ACTIV-2 COVID-19 Symptom Diary (ACSD).

BACKGROUND: Patient-reported outcome measures are needed to assess the impact of treatments for COVID-19 on symptoms. The ACTIV-2 COVID-19 Symptom Diary (ACSD) is being used in the ongoing Accelerating COVID-19 Therapeutic Interventions and Vaccines-2 (ACTIV-2) platform clinical trial. The purpose of the current study was to conduct qualitative interviews to assess content validity of the ACSD. METHODS: Interviews were conducted with adults who had tested positive for SARS-CoV-2. The ACSD begins with global items, followed by a symptom checklist. Each interview began with concept elicitation focusing on participant experiences with COVID-19. Then, participants completed the ACSD, and cognitive interviews were conducted to evaluate the questionnaire. Interviews were recorded, transcribed, and coded following a qualitative content analysis. For the qualitative analysis, a coding dictionary was developed with a list of all potential codes and instructions for how the codes should be applied and combined. RESULTS: Interviews were conducted with 30 participants (mean age = 39 years; 57% female; 17% Latinx; 17% Black/African American; 40% meeting at least one criterion for classification as high risk of progression to severe COVID-19). Commonly reported symptoms included fatigue (reported by 100% of the sample), body pain/muscle pain/aches (87%), headaches (87%), cough (83%), loss of smell (73%), shortness of breath/difficulty breathing (70%), and chills (70%). The 13 symptoms most commonly reported in this study are included in the ACSD. After completing the ACSD, participants consistently reported that it was clear and easy to complete, and all items were generally interpreted as intended. Based on participants' input, the ACSD was edited slightly after the first 13 interviews, and the revised version was used for the final 17 interviews. Two additional items assessing "brain fog" and dizziness were recommended for addition to the ACSD in future research. CONCLUSIONS: This qualitative study supports the content validity of the ACSD for assessment of COVID-19 symptoms. Quantitative research with larger samples will be needed to examine the questionnaire's measurement properties.

This study focused on the ACTIV-2 COVID-19 Symptom Diary (ACSD), a questionnaire that assesses symptom severity of COVID-19. The ACSD begins with global items assessing overall symptom severity, followed by a symptom checklist focusing on individual symptoms. Interviews were conducted with 30 adults who had tested positive for COVID-19. The patients reported their experiences with COVID-19, completed the ACSD, and provided their opinions about the ACSD. Based on input from these patients, the ACSD appears to be clear and easy to complete, and it includes the most common and important symptoms of COVID-19. The ACSD was edited for clarity, and "brain fog" and dizziness were recommended additions for future research. This study suggests that the ACSD is a useful questionnaire for assessment of COVID-19 symptoms in clinical studies. Studies like this are important for ensuring that symptoms are measured appropriately and accurately in clinical trials. Future research with larger samples will be needed to further examine the questionnaire.

The effect of reduced CP, synthetic amino acid supplemented diets on growth performance and nutrient excretion in wean to Finish swine.

Mixed sex pigs (n = 720) were placed in 12 rooms (Purdue Swine Environmental Research Building) to measure the effect of reduced crude protein (CP), amino acid (AA)-supplemented diets on growth and the carcass. Pigs were blocked by body weight (BW) and gender and allotted to room and pen (10 mixed-sex pigs/pen). Pigs were fed a nine-phase, wean-finish program. Control pigs consumed corn-soybean meal-distiller's dried grains with solubles (DDGS) diets containing no to minimal (Met) synthetic AA. The 2X diet was formulated to meet the seventh most-limiting AA, and balanced using synthetic AAs to meet all AA needs. The 1X diet was formulated to meet a CP value halfway between the control and 2X diet, and also balanced using synthetic AAs to meet all AA needs. Diets were formulated to identical net energy concentrations and balanced to meet standard ileal digestible NRC 2012 AA requirements. Pit vacuum samples were collected at the end of each growth phase for analyses of nitrogen, C and dry matter (DM). Pigs fed the Control and 1X diet grew faster (P < 0.005), had greater gain:feed (P < 0.001), and were heavier at market (P < 0.001) than animals fed the 2X diet. No consistent effects of diet were observed on average daily feed intake. Carcass data were analyzed for sex, diet and sex*diet effects. Reductions in dietary CP resulted in a linear reduction in ammonium nitrogen excretion per kg of BW gain in Nursery (P < 0.001) and Grow-Finish (P < 0.001) phases. Reductions in dietary CP, with synthetic AA supplementation resulted in a linear reduction in total nitrogen excreted per kg BW gain in the Grow-Finish phase (P < 0.001) and overall (P < 0.001). Total mineral excretion per kg gain was reduced in pigs fed 1X and 2X diets compared with control-fed pigs (P < 0.005). Reductions in dietary CP of ~3 and 5%-units from wean-finish result in reductions of total N excretion of 11.7 and 24.4%, respectively. Reduced performance and carcass characteristics observed in pigs fed the 2X diets indicates an inaccurate estimate of NRC 2012 AA requirements or ratios to lysine in a low CP diet.

Mammary Development in Gilts at One Week Postnatal Is Related to Plasma Lysine Concentration at 24 h after Birth, but Not Colostrum Dose.

Perinatal nutrition affects future milk production. The number of mammary epithelial cells affect milk production capacity. Therefore, it was hypothesized that the level of colostrum intake affects the proliferation rate and the total number of mammary epithelial cells in the gland. The ratio of newly synthesized protein to newly synthesized DNA reflects the relative amount of cellular differentiation to cell division. The study objective was to determine the relationship between the level of colostrum intake and 24 h-level of circulating amino acid, glucose and insulin with mammary parenchyma histological features, cell division and protein synthesis over the first week postnatal. One of two standardized doses of a homogenate colostrum sample, 10% (n = 8) and 20% (n = 8) of birth bodyweight, was fed to gilts over the first 24 h postnatal. Gilts were administered deuterium oxide immediately after birth and daily to label newly synthesized DNA and proteins. Gilts were euthanized on postnatal day seven, and DNA and protein were isolated from mammary parenchyma. DNA and protein fractional synthesis (f) and fractional synthetic rate (FSR) were calculated using mass isotopomer distribution analysis. The ratio of protein f and FSR to DNA f and FSR were calculated and used to indicate the relative amounts of differentiation to cell division. Mammary morphological development was also analyzed by measuring the parenchymal epithelial area and the stromal and epithelial proliferation index on postnatal day seven. Colostrum dose was not related to any of the variables used to evaluate mammary development. However, plasma lysine levels at 24 h postnatal were positively related to average daily gain (ADG; r = 0.54, p = 0.05), DNA f (r = 0.57; p = 0.03) and DNA FSR (r = 0.57; p = 0.03) in mammary parenchyma. Plasma lysine was inversely related to the ratio of protein to DNA f and FSR (r = -0.56; p = 0.04). ADG was related to the parenchymal epithelial area and DNA and protein f and FSR (p < 0.05). These relationships support the idea that the nutritional environment affects early mammary development and that higher lysine levels in the perinatal period favored a greater degree of cell division versus differentiation in mammary of neonatal pigs and thus, warrant further investigations.

Biomarkers predictive of long-term fertility found in vaginal lipidome of gilts at weaning.

A marker indicative of the fertility potential of replacement gilts early in development would decrease culling rates in the sow herd, improve sow herd reproductive efficiency, and reduce production costs. The objective of this study was to determine if vaginal lipid profiles at 21 d postnatal (PN) could predict sow reproductive performance. Vaginal swabs of the anterior vagina were taken at 21 ± 4 d PN from gilts born on a commercial sow production facility for lipidomic analysis. Animals were followed prospectively for 2 yr and assigned to reproductive performance categories based on the observation of estrus or piglets weaned per sow per year (PSY) across two farrowings. Lipids were extracted from cellular material collected with swabs taken from high fertility (HF; n = 28; ≥26 PSY) and infertile (IF; n = 34; no estrus, no pregnancy) animals, and multiple reaction monitoring profiling was used for lipidome analysis. The relative abundance of arachidonic acid (C20:4) and docosahexaenoic acid (C22:6) was lower (P < 0.05) in IF gilts than HF gilts, whereas the abundance of the free fatty acids such as cerotic (C26:0), ximenic (C26:1), and nonadecanoic (C19:0) acids was greater (P < 0.05) in IF gilts. Additionally, eicosapentaenoic acid (C20:5), a precursor of prostaglandins, was higher (P < 0.05) in IF gilts. The perspective of having a panel of lipids captured with vaginal swabs at weaning that can predict the reproductive efficiency of gilts shows promise and warrants future research in this area.

Effects of induction on the farrowing process and piglet blood parameters at the time of farrowing.

Historically, sows have been induced to farrow using prostaglandin followed by an injection of oxytocin 24 h later. Benefits of induction can include decreased rate of stillbirths, dystocia, and postnatal mortality along with increasing the likelihood of farrowings being attended. Several studies have indicated that oxytocin administration may negatively impact fetal oxygen supply during parturition, potentially from umbilical cords breaking prior to birth, resulting in increased preweaning mortality. Therefore, the objective of this study was to determine if various induction protocols impact umbilical cord breakage and fetal blood parameters at birth. Fifty-eight primiparous and multiparous sows were assigned to one of three treatments: no induction (NO; n = 24) or 2 cc prostaglandin administered on day 114 of gestation followed by either 1 cc of oxytocin 24 h later (OXY24; n = 13) or 0.5 cc of oxytocin at 6 and 12 h after prostaglandin (OXY6; n = 21). Details of the farrowing process were recorded, and umbilical cord blood was collected from piglets at birth and evaluated on an iSTAT machine using an Abbott EC8+ test cartridge. There were no differences in total born, number born alive, stillborns, mummies, or assistance needed during farrowing. Induced sows were more likely to farrow by day 115 compared to naturally farrowing sows (P = 0.02). Sows in the OXY24 treatment tended to have longer farrowings when compared to both NO and OXY6 (4.8 vs. 3.6 vs. 3.9 h; P = 0.09). Colostrum from OXY6 sows tended to have a greater amount of lactose present than NO and OXY24 (P = 0.05). Colostrum from sows with longer gestation lengths had a higher percentage of fat (P = 0.03). Piglets born from NO sows had higher base excess, total carbon dioxide, and glucose, which suggests that these piglets had prolonged moments of asphyxiation (P < 0.01). OXY24 piglets had the lowest blood pH which is indicative of hypoxic birthing conditions (P < 0.01). Preweaning mortality was driven largely by a low birth weight coupled with low colostrum intake (P = 0.03). All piglets, regardless of treatment, displayed signs of stress during farrowing. Induction did not influence preweaning mortality but has the potential to decrease the incidence by increasing attended farrowings.

Shotgun proteome analysis of seminal plasma differentiate boars by reproductive performance.

There is a need to identify subfertile boars before they enter into the breeding herd. Seminal plasma proteins are essential for normal sperm function and transport and play an important role in fertilization. The objective of this study was to use liquid chromatography tandem mass spectrometry for shotgun proteome analysis to investigate whether differences in boar fertility phenotype can be differentiated by seminal plasma protein abundance. Following 50 breedings, boars were categorized into one of four phenotypes: high farrowing rate and total born (HFHB; n = 9), high farrowing rate with low total born (HFLB; n = 10), low farrowing rate and total born (LFLB; n = 9), and low farrowing rate with high total born (LFHB; n = 4) that were distinct (p < 0.05) from each other by these variables. There were 506 proteins measured in at least one sample across all animals. There were 245 high confidence proteins and 56 were differentially abundant between the high fertility phenotype (HFHB) and at least one of the three subfertile groups. Findings support that seminal plasma protein profiles are distinct between boars with different fertility phenotypes.

Evaluation of on-farm indicators of gilt reproductive performance potential at 21 days of age.

Selection of replacements for the sow herd is one of the most important facets in swine production. Although our current methods of selection are effective, there is still a large amount of variation in sow reproductive performance traits such as pigs per sow per year (PSY). Therefore, the objective of this study was to determine if on-farm phenotypic traits at 21 d postnatal (PN) or perinatal environmental factors could predict sow reproductive performance. Data were prospectively collected from 2,146 gilts born on a commercial sow production facility and included birth and weaning weights, vulva length and width at 21 d PN, birth and nursing litter size, days nursed, average daily gain from birth to weaning, and age at first estrus. Of the initial animals, 400 (17%) were selected for the sow herd, 353 remained after removal of animals culled for non-reproductive reasons. Animals were assigned to 1 of 5 reproductive performance categories based on observation of estrus or pigs per sow per year (PSY) across two farrowings: High Fertility (HF; 23%; n = 82; ≥26 PSY), Middle Fertility (MF2; 12%; n = 43; 20-25 PSY), Low Fertility (MF3; 15%; n = 54; <20 PSY), Infertile-Estrus (IFe; 10%; n = 36; estrus, no pregnancy), and Infertile-No Estrus (IFno; 39%; n = 138; no estrus, no pregnancy). Generalized linear model analysis indicated vulva width (P = 0.03) was related to PSY, however, it only explained 1.5% of the total variation in PSY. To determine if preweaning variables were predictive of gilt fertility outcome, animals were grouped as those that became pregnant (n = 179) or not (n = 174). Vulva width tended to be greater in fertile animals versus infertile (P = 0.07). Binomial regression analysis revealed a positive relationship between vulva width and gilt fertility, however, this relationship is not strong enough to make sow herd selection decisions.

Colloidal stability of citrate and mercaptoacetic acid capped gold nanoparticles upon lyophilization: effect of capping ligand attachment and type of cryoprotectants.

For various applications of gold nanotechnology, long-term nanoparticle stability in solution is a major challenge. Lyophilization (freeze-drying) is a widely used process to convert labile protein and various colloidal systems into powder for improved long-term stability. However, the lyophilization process itself may induce various stresses resulting in nanoparticle aggregation. Despite a plethora of studies evaluating lyophilization of proteins, liposomes, and polymeric nanoparticles, little is known about the stability of gold nanoparticles (GNPs) upon lyophilization. Herein, the effects of lyophilization and freeze-thaw cycles on the stability of two types of GNPs: Citrate-capped GNPs (stabilized via weakly physisorbed citrate ions, Cit-GNPs) and mercaptoacetic acid-capped GNPs (stabilized via strongly chemisorbed mercaptoacetic acid, MAA-GNPs) are investigated. Both types of GNPs have similar core size and effective surface charge as evident from transmission electron microscopy and zeta potential measurements, respectively. Plasmon absorption of GNPs and its dependence on nanoparticle aggregation was employed to follow stability of GNPs in combination with dynamic light scattering analysis. Plasmon peak broadening index (PPBI) is proposed herein for the first time to quantify GNPs aggregation using nonlinear Gaussian fitting of GNPs UV-vis spectra. Our results indicate that Cit-GNPs aggregate irreversibly upon freeze-thaw cycles and lyophilization. In contrast, MAA-GNPs exhibits remarkable stability under the same conditions. Cit-GNPs exhibit no significant aggregation in the presence of cryoprotectants (molecules that are typically used to protect labile ingredients during lyophilization) upon freeze-thaw cycles and lyophilization. The effectiveness of the cyroprotectants evaluated was on the order of trehalose or sucrose > sorbitol > mannitol. The ability of cryoprotectants to prevent GNPs aggregation was dependent on their chemical structure and their ability to interact with the GNPs as assessed with zeta potential analysis.