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Introduction: Coronavirus disease 2019 (COVID-19) has impacted researchers' ability to continue to deliver HIV prevention and treatment interventions face to face. Although telehealth has been an important strategy to maintain research operations during the current pandemic, participants at increased risk of or living with HIV are often at higher risk of also experiencing poverty, housing instability, and other challenges that may present obstacles to successful remote delivery. Methods: We provide descriptions of remote adaptations to two randomized controlled efficacy trials of behavioral interventions for primary and secondary HIV prevention with descriptive enrollment and retention data. Results and Conclusions: Best practices for implementing telemedicine and e-health procedures are discussed, including procedures for addressing remote participation barriers (economic, health literacy, etc.) and other challenges, such as building rapport and staff support (NCT03092531 and NCT03175159).

OBJECTIVES: Acceptance and Commitment Therapy (ACT) is a third-wave behavioral and cognitive therapy that increases psychological flexibility through mindfulness, acceptance, and value-driven behavior change. ACT has been successfully used to inform a variety of health interventions. Using non-therapists to deliver ACT-based behavioral health interventions offers an opportunity to provide cost efficient and integrated care, particularly among underserved populations experiencing barriers to mental health care, such as inadequate insurance, mental health stigma, and provider shortages. This systematic review aims to: 1) identify ACT-informed behavioral health interventions delivered by laypeople and 2) review the specific characteristics of each intervention including number and duration of sessions, delivery modality, interventionist training, and intervention outcomes. METHODS: Two databases (PubMed and PsycINFO) were systematically searched for relevant literature. To further identify relevant studies, references of included manuscripts were checked, the Association for Contextual Behavioral Science’s webpage was examined, and an email was sent to the ACBS Health Special Interest Group listserv. Study abstracts and full texts (in English) were screened, resulting in 23 eligible articles describing 19 different interventions. RESULTS: A total of 1,781 abstracts were screened, 76 were eligible for full-text review, and 23 were included in a narrative synthesis. There were 19 unique interventions identified and delivered by the following: general healthcare workers (n= 7), trained researchers (n = 5), women/mothers (n= 2), municipal workers (n= 2), and teachers (n = 3). Eleven studies were RCTs and eight utilized alternative study designs. Study quality varied, with two rated as high risk for bias and eight rated to have some concerns. Target populations included clinical and non-clinical samples. There was some consistency in the effects reported in the studies: increases in pain tolerance, acceptance, and identifying and engaging in value driven behavior, improvements in cognitive flexibility, and reductions in psychological distress. CONCLUSIONS: Findings suggest that ACT interventions can be successfully delivered by a variety of laypeople and effectively address psychological distress and increase health behaviors.

Heat stress affects oocyte developmental competence and is a major cause of reduced fertility in heat stressed cattle. Negative effects of heat stress on the oocyte have been observed at morphological, biochemical and developmental levels. However, the mechanisms by which heat stress affects the oocyte at the transcriptional and epigenetic levels remain to be further elucidated. Here we aimed to investigate the effect of heat stress on oocyte quality, transcriptomic profiles and DNA methylation of oocytes collected through the transition from spring to summer under Louisiana conditions. Summer season resulted in a lower number of high quality oocytes obtained compared to the spring season. There was no difference in in vitro maturation rates of oocytes collected during spring as compared to summer. RNA sequencing analysis showed that a total of 211 and 92 genes were differentially expressed as a result of heat stress in GV and MII oocytes, respectively. Five common genes (E2F8, GATAD2B, BHLHE41, FBXO44, and RAB39B) were significantly affected by heat in both GV and MII oocytes. A number of pathways were also influenced by heat stress including glucocorticoid biosynthesis, apoptosis signaling, and HIPPO signaling in GV oocytes, and Oct4 pluripotency, Wnt/beta-catenin signaling, and melatonin degradation I in MII oocytes. In addition, fluorescent immunocytochemistry analysis showed no difference in global levels of DNA methylation and DNA hydroxymethylation at either the GV or MII stage between spring and summer oocytes. The results of this study contribute to a better understanding of the effect of heat stress on the molecular mechanisms altered in bovine oocytes.

This chapter turns attention to how respondents asses the short- and long-term viability of Clarksdale's blues tourism strategy. Respondents were frustrated and skeptical. In their view, the blues campaign had done very little to improve the stakes and outlook of the town's black residents. Jobs had not come. The town's infrastructure was deplorable. Race relations were contentious. Respondents were also cautiously hopeful—vigilant.

This chapter chronicles the establishment and growth of blues tourism in the Mississippi Delta, with attention to the town of Clarksdale. Foster argues that this blues development agenda was established to address the region's declining demographic and economic profile. By the end of the chapter, a tension emerges—between the viability of blues tourism as an economic boon and the question of which communities benefit the most from the growing revenue stream.

Chapter two chronicles the ways that Black residents of Clarksdale define, talk about, and "use" the blues. Foster identifies three elements of blues talk—definitions that center personal memories and past experiences, definitions that draw distinctions between who can and cannot sing/live the blues, and definitions that celebrate a shared sense of group belonging. Foster considers the role that these definitions—and the feelings they engender—play in process of racial identity and social boundary making.

How do you love and not like the same thing at the same time? This was the riddle that met Mississippi writer B. Brian Foster when he returned to his home state to learn about Black culture and found himself hearing about the blues. One moment, Black Mississippians would say they knew and appreciated the blues. The next, they would say they didn’t like it. For five years, Foster listened and asked: “How?” “Why not?” “Will it ever change?” This is the story of the answers to his questions. In this illuminating work, Foster takes us where not many blues writers and scholars have gone: into the homes, memories, speculative visions, and lifeworlds of Black folks in contemporary Mississippi to hear what they have to say about the blues and all that has come about since their forebears first sang them. In so doing, Foster urges us to think differently about race, place, and community development and models a different way of hearing the sounds of Black life, a method that he calls listening for the backbeat.

Capacity-oriented approaches to health interventions seek to empower the target population or community to manage the health issue themselves using resources they can control. Positive deviance, resilience and asset-based approaches are three such methods of developing and implementing health interventions. This study aimed to review the efficacy of interventions explicitly applying these methods in addressing childhood obesity using adiposity as the primary outcome, measured by standardized body mass index. The search strategy was developed and implemented across four electronic databases. Of the 181 records identified and screened, 11 studies were identified as using a capacity-oriented approach overall. Asset-based approaches (n = 8 studies) consisted of 47 880 participants, positive deviance (n = 2 studies) consisted of 781 participants, and resilience-based interventions (n = 1 study) consisted of 35 participants. The asset-based approaches were mixed, with three of the eight studies showing a significant reduction in adiposity, while the other five did not find a difference. The positive deviance and resilience-based studies showed signs of efficacy in reducing adiposity. There was significant design heterogeneity across studies, and varied interpretations and definitions of the approaches were used. Further work should attempt to achieve some consensus on the use of these approaches to facilitate comparison and advance the science of capacity-oriented interventions for childhood obesity.

Path planning in additive manufacturing (AM) processes has an impact on the thermal histories experienced at discrete locations in simple and complex AM structures. One component of path planning in directed energy deposition is the time required for the laser or heat source to return to a given location to add another layer of material. As structures become larger and more complex, the length of this interlayer dwell time can significantly impact the resulting thermal histories. The impact of varying dwell times between 0 and 40 seconds on the microstructural and mechanical properties of Inconel® 625 and Ti-6Al-4V builds has been characterized. Even though these materials display different microstructures and solid-state phase transformations, the addition of an interlayer dwell generally led to a finer microstructure in both materials that impacted the resulting mechanical properties. With the addition of interlayer dwell times up to 40 seconds in the Inconel® 625 builds, finer secondary dendrite arm spacing values, produced by changes in the thermal history, correspond to increased yield and tensile strengths. These mechanical properties did not appear to change significantly, however, for dwell times greater than 20 seconds in the Inconel® 625 builds, indicating that longer dwell times have a minimal impact. The addition of interlayer dwell times in Ti-6Al-4V builds resulted in a slight decrease in the measured alpha lath widths and a much more noticeable decrease in the width of prior beta grains. In addition, the yield and tensile values continued to increase, nearly reaching the values observed in the rolled plate substrate material with dwell times up to 40 seconds.

Hypophosphatasia (HPP) is an inherited disorder that affects bone and tooth mineralization characterized by low serum alkaline phosphatase. HPP is caused by loss-of-function mutations in the ALPL gene encoding the protein, tissue-nonspecific alkaline phosphatase (TNSALP). TNSALP is expressed by mineralizing cells of the skeleton and dentition and is associated with the mineralization process. Generalized reduction of activity of the TNSALP leads to accumulation of its substrates, including inorganic pyrophosphate (PP

Hypophosphatasia (HPP) is an inherited disorder that affects bone and tooth mineralization characterized by low serum alkaline phosphatase. HPP is caused by loss-of-function mutations in the ALPL gene encoding the protein, tissue-nonspecific alkaline phosphatase (TNSALP). TNSALP is expressed by mineralizing cells of the skeleton and dentition and is associated with the mineralization process. Generalized reduction of activity of the TNSALP leads to accumulation of its substrates, including inorganic pyrophosphate (PPi) that inhibits physiological mineralization. This leads to defective skeletal mineralization, with manifestations including rickets, osteomalacia, fractures, and bone pain, all of which can result in multi-systemic complications with significant morbidity, as well as mortality in severe cases. Dental manifestations are nearly universal among affected individuals and feature most prominently premature loss of deciduous teeth. Management of HPP has been limited to supportive care until the introduction of a TNSALP enzyme replacement therapy (ERT), asfotase alfa (AA). AA ERT has proven to be transformative, improving survival in severely affected infants and increasing overall quality of life in children and adults with HPP. This chapter provides an overview of TNSALP expression and functions, summarizes HPP clinical types and pathologies, discusses early attempts at therapies for HPP, summarizes development of HPP mouse models, reviews design and validation of AA ERT, and provides up-to-date accounts of AA ERT efficacy in clinical trials and case reports, including therapeutic response, adverse effects, limitations, and potential future directions in therapy.

Reduced reproductive performance is one of the main effects caused by heat stress in cattle. Its negative effects have been observed at the transcriptional, biochemical, morphological, and developmental levels on the oocyte and embryo. There are no studies evaluating the effect of heat stress on the epigenetic profile of bovine oocytes and early embryos. The objective of this study was to evaluate the effect of invivo and invitro heat stress on DNA methylation and DNA hydroxymethylation in bovine MII oocytes, pronuclear, and 2- to 4-cell stage embryos. Seven Bos taurus crossbred nonpregnant, non-lactating beef cows located in Saint Gabriel, Louisiana (30.269746, −91.103357) were used for oocyte collection. Dominant follicle removal was performed 5 days before oocyte collection. Cumulus–oocyte complexes were collected by ovum pickup from follicles >2 mm. Samples were collected during the summer (August) and winter (February) (5 collections each). Three treatments were utilised: invivo heat stress (August samples), invitro heat stress (February samples subjected to 41°C during the first 12 h of IVM and then to 38.5°C during the next 12 h of IVM), and control (February samples IVM at 38.5°C). All oocytes collected per treatment were assigned to 3 developmental stages: MII oocytes, pronuclear, and 2- to 4-cell stage embryos. Embryos were obtained through standard IVF. DNA methylation and DNA hydroxymethylation was assessed by fluorescence immunohistochemistry utilising primary antibodies against 5′-methylcytosine and 5′-hydromethylcytosine and secondary antibodies Alexa Fluor 488 and Alexa Fluor 546, respectively. Samples were visualised with a fluorescence deconvolution microscope, and immunofluorescence data were expressed as corrected relative fluorescence per nucleus. Results were analysed by the Type III test of fixed effects and Tukey media separation utilising the Proc Glimmix of SAS 9.4 (P

Embryo vitrification involves exposure to high concentrations of cryoprotectants and osmotic stress during cooling and warming in the cryopreservation process. Many of these factors can potentially affect gene expression. In this study, invitro-produced bovine embryos at the blastocyst stage were subjected to vitrification. Four recipients each were used for transferring non-vitrified (n=80) and vitrified (n=80) embryos. A total of 12 non-vitrified and 9 vitrified viable day-14 (D14) embryos were recovered by uterine flushing. RNA-seq analysis of the whole embryo or isolated trophectoderm (TE) from vitrified and fresh recovered D14 embryos revealed a total of 927 and 4376 genes with changed expression in embryos and TE isolates, respectively, as a result of vitrification. In addition, we found 671 and 61 genes commonly up- or downregulated in both vitrified whole embryos and TE. Commonly upregulated pathways by vitrification included epithelial adherens junctions, sirtuin signalling, germ cell–sertoli cell junction, ATM signalling, NER and protein ubiquitination pathways. The commonly downregulated pathways included EIF2 signalling, oxidative phosphorylation, mitochondrial dysfunction, regulation of eIF4 and p70S6K signalling and mTOR signalling pathways. Our analysis identified specific pathways and implicated specific gene expression patterns affecting embryo developmental competence that are important to cryopreservation.

Introduction In developed countries, cardiovascular disease (CVD) is the leading cause of death. Identification and mitigation of CVD risk factors has been the chief strategy for prevention of CVD. To this end, the American Heart Association (AHA) and American College of Cardiology (ACC) suggest metrics of ideal cardiovascular (CV) health in seven domains and have published guidelines encouraging use of the “Simple Seven” to improve CV health. Sleep is notably absent from the Simple Seven despite substantial epidemiological evidence of its importance for CV health. We sought to measure the correlation of sleep parameters with CVD risk estimate by Framingham risk score (FRS). Methods Subjects prospectively enrolled in a CV Health Registry provided data for FRS, the seven domains of the “Simple Seven” including blood pressure, physical activity, total cholesterol, diet, weight, smoking, and glucose level, and completed validated questionnaires: Pittsburgh Sleep Quality Index (PSQI) and Berlin Questionnaire (BQ) for sleep apnea. Pearson correlations were analyzed between FRS and the “Simple Seven” as well as for the sleep questionnaires for comparison. Results Among the 646 subjects (mean age 55.4 ±9.9y, 43% men, 399W, 190B, 26H, 12A, 19other), FRS correlated substantially with blood glucose (R=0.398, p Conclusion This study suggests that sleep parameters, specifically BQ and PSQI, may contribute to risk assessment of CVD while also offering actionable information for improving CV health. Support

Capacity-oriented approaches to health interventions seek to empower the target population or community to manage the health issue themselves using resources they can control. Positive deviance, resilience and asset-based approaches are three such methods of developing and implementing health interventions. This study aimed to review the efficacy of interventions explicitly applying these methods in addressing childhood obesity using adiposity as the primary outcome, measured by standardized body mass index. The search strategy was developed and implemented across four electronic databases. Of the 181 records identified and screened, 11 studies were identified as using a capacity-oriented approach overall. Asset-based approaches (n = 8 studies) consisted of 47 880 participants, positive deviance (n = 2 studies) consisted of 781 participants, and resilience-based interventions (n = 1 study) consisted of 35 participants. The asset-based approaches were mixed, with three of the eight studies showing a significant reduction in adiposity, while the other five did not find a difference. The positive deviance and resilience-based studies showed signs of efficacy in reducing adiposity. There was significant design heterogeneity across studies, and varied interpretations and definitions of the approaches were used. Further work should attempt to achieve some consensus on the use of these approaches to facilitate comparison and advance the science of capacity-oriented interventions for childhood obesity.

Background: Addressing suboptimal breastfeeding initiation and duration rates is a priority in the United States. To address challenges to improving these rates, the voices of the providers who work with breastfeeding mothers should be heard. Research aim: The purpose of this study was to explore lactation consultants’ perceived barriers to managing early breastfeeding problems. Methods: This qualitative study was conducted with a grounded theory methodological approach. In-depth interviews were conducted with 30 International Board Certified Lactation Consultants across Florida. Lactation consultants were from a range of practice settings, including hospitals, Special Supplemental Nutrition Program for Women, Infants, and Children clinics, private practice, and pediatric offices. Data were digitally recorded, transcribed, and analyzed in Atlas.ti. Results: A range of barriers was identified and grouped into the following categories/themes: indirect barriers (social norms, knowledge, attitudes); direct occupational barriers (institutional constraints, lack of coordination, poor service delivery); and direct individual barriers (social support, mother’s self-efficacy). A model was developed illustrating the factors that influence the role enactment of lactation consultants in managing breastfeeding problems. Conclusion: Inadequate support for addressing early breastfeeding challenges is compounded by a lack of collaboration among various healthcare providers and the family. Findings provide insight into the professional management issues of early breastfeeding problems faced by lactation consultants. Team-based, interprofessional approaches to breastfeeding support for mothers and their families are needed; improving interdisciplinary collaboration could lead to better integration of lactation consultants who are educated and experienced in providing lactation support and management of breastfeeding problems.

In current in vitro embryo production practices, method of oocyte recovery tends to be based primarily on the personal preference of the operators and availability of resources rather than on any practical understanding of the best practices for retrieving oocytes of the greatest competence for embryo production. There is little consensus on the effects of either postmortem follicular atresia or exogenous hormone-stimulated follicle development on the quality of oocytes, but it is expected that both interventions have effects. Cumulus-oocyte complexes were collected from beef cattle using 3 methods: unstimulated ovum pickup (OPU), OPU following superstimulation (FSH), and oocyte aspiration from postmortem-recovered ovaries (PM). Each collection method was repeated 3 times, with 25 individuals used in each. There were no interventions in place before oocyte recovery in either OPU or PM. In FSH, cows underwent dominant follicle removal and were treated with 200mg of FSH in 6 doses before OPU. The majority of oocytes (66 OPU, 80 FSH, and 160 PM) were fertilized and cultured, with a portion being reserved for transmission electron microscopy (2 per treatment at each of germinal vesicle and metaphase II) and a portion being reserved for a complementary study. However, oocytes collected by OPU following FSH had notable ultrastructure differences from those collected by other methods, suggesting a potential effect of follicle stimulation on both the development and cytoplasmic maturation of oocytes. Although oocytes from all recovery methods and at both maturation levels exhibited mitochondria primarily in the hooded conformation, both OPU and FSH oocytes, at germinal vesicle and metaphase II, had several anomalous mitochondria exhibiting a lobulated, budding-type shape. This is potentially indicative of these oocytes having failed to complete the mitochondrial division phase before recovery. An average of 11.6, 10.6, and 11.9 oocytes per individual were recovered from PM, FSH, and OPU, respectively, resulting in oocyte recovery of 55, 61, and 57% of aspirated follicles, respectively (P>0.05). Fewer oocytes from OPU developed to morulas compared with other recovery methods (chi-square; P

Cattle under the effect of heat stress have reduced fertility, with negative effects on the oocyte observed at the morphological, biochemical, transcriptional and developmental levels. There are no studies evaluating the effect of heat stress on the epigenetic profile of bovine oocytes, which plays a fundamental role in the regulation of gamete development. The objective of this study was to evaluate the effect of in vivo heat stress during the spring to summer transition on DNA methylation and DNA hydroxymethylation of bovine oocytes at the germinal vesicle (GV) and metaphase II (MII) stages. Ten Bos taurus crossbred nonlactating beef cows located at Saint Gabriel, Louisiana, USA (30°16′11.1″ N, 91°06′12.1″ W), were used for oocyte collection once monthly from April to August. Dominant follicle removal was performed 5-7 days before oocyte collection. Cumulus-oocyte complexes were collected through ovum pick-up from follicles >2 mm. Germinal vesicle (GV)-stage oocytes (50% of total obtained per cow) were subjected to a standard bovine in vitro maturation protocol to obtain metaphase II (MII) stage oocytes. The DNA methylation and DNA hydroxymethylation of GV and MII oocytes was assessed by fluorescence immunohistochemistry utilising primary antibodies against 5′-methylcytosine and 5′-hydromethylcytosine. Secondary antibodies utilised were Alexa Fluor 488 goat anti-mouse IgG and Alexa Fluor 546 donkey anti-rabbit IgG. Oocytes were visualised utilising a fluorescence deconvolution microscope and immunofluorescence data were expressed as corrected relative fluorescence per nucleus. The polar body was not included for fluorescence quantification when evaluating MII stage oocytes. Results (least squares means ± standard error) were evaluated as cold months (April and May) and hot months (June, July, and August). Results were analysed by the type III test of fixed effects and Tukey media separation utilising Proc Glimmix of SAS 9.4 (P

It is known that animals under the effect of heat stress present reduced fertility. We aimed to investigate the effect of heat stress on the developmental competence and global gene expression profile of oocytes through the transition from spring to summer under Louisiana conditions. Oocytes were collected from 6 crossbred, non-lactating cows once a month from May to July. Temperature and humidity indexes for May and July were 72.48 and 78.06, respectively. An index above 75 indicates that cows are under heat stress. All cows underwent dominant follicle removal, and then 7 days later, ovum pickup was performed to aspirate germinal vesicle (GV) oocytes. Half of the oocytes were processed for RNA-seq as GV, and half were matured in vitro to metaphase II (MII). Smart-seq protocol was followed to prepare RNA sequencing libraries from a pool of 4 oocytes (GV n=6; MII n=6). Sequencing reads were pre-filtered and aligned to the bovine genome, and gene expression values were calculated as transcripts per million. Genes were deemed differentially expressed between different conditions if they showed a false discovery rate P-value

Since the late 1970’s, extensive testing has been conducted at Gramercy to remove high molecular weight organic compounds (humates) from Bayer process liquors. Early research work tested the removal of humates using the Giulini process (Dolime) , and by the use of organic polymeric additives.

Zona pellucida hardening is a natural process that occurs after oocyte fertilization to prevent polyspermic fertilization and to protect embryonic development. Pre-fertilization hardening of the zona pellucida, however, decreases fertilization rates. Cryoprotectants have also been shown to negatively affect fertilization rates, one possible mechanism of which being through zona hardening. This experiment was conducted to determine the effect of different cryoprotectants on hardening of the zona pellucida of mature bovine oocytes. Oocytes were collected by ovum pick-up (OPU) by transvaginal ultrasound guided aspiration (TUGA) from mixed-breed cows. After collection, oocytes were randomly assigned to 3 cryoprotectant treatment groups: dimethyl sulfoxide (DMSO), glycerol, or PBS (control). Drops (50 µL) of each vitrification solution were placed under mineral oil. Vitrification solution 1 (VS1) contained 10% ethylene glycol (EG), either 10% DMSO or glycerol, and 0.5 M sucrose. Vitrification solution 2 (VS2) contained 20% EG, 20% DMSO or glycerol, and 0.5 M sucrose. All oocytes were held in VS1 for 5 min before being transferred to VS2 for 45 s. All oocytes were washed in a common dilution solution (80% PBS, 20% calf serum, 0.025 M sucrose) for 5 min. Next, oocytes were moved to 50-µL drops of protease solution (0.1% protease) under mineral oil. Control oocytes were held in PBS for ~10 min before entering the protease solution to represent the same period as the vitrification procedure. The oocytes were observed until the zonae pellucidae were completely digested and times were recorded for each oocyte. This experiment included 4 replicates with a total of 88 oocytes used, 32 each in DMSO and glycerol and 24 in PBS. The data were analysed using ANOVA. The DMSO group had the lower mean zona digestion time out of the 2 cryoprotectants at 15.75 min and glycerol had the highest mean digestion time at 19.3 min. The control group (PBS) had the lowest mean of the 3 treatments at 12.7 min. The differences between DMSO and glycerol, and between DMSO and PBS were not significant (P = 0.0654 and 0.1073, respectively). However, both glycerol versus PBS and the average of DMSO and glycerol versus PBS were significantly different (P-value = 0.0053 and 0.0119, respectively). These results suggest that glycerol hardens the zona pellucida more than DMSO or PBS; however, there is not enough evidence to determine whether DMSO hardens the zona pellucida compared with PBS. This would suggest that, in relation to zona hardening and ensuring proper fertilization, glycerol-based cryoprotectants may be a better option than DMSO-based ones. Further, these results may be important in embryo vitrification as zona hardening may prevent blastocyst hatching, suggesting that glycerol-based cryoprotectants should be investigated as the optimal cryoprotectant here also.

Anti-mullerian hormone (AMH) has become an important tool for the selection of donors with high antral follicle count and high superovulatory response for in vivo or in vitro embryo production programs. The serum concentration of AMH is becoming a suitable selection marker because it presents minimal variation during the oestrous cycle and is repeatable during multiple cycles. It is known that heat stress results in decreased production of reproductive steroid and protein hormones in stressed animals. Importantly, the effect of heat stress on the serum concentration of AMH is still unknown. The objective of this experiment was to evaluate the antral follicle count and AMH serum concentration levels during the spring to summer transition to assess the effect of heat stress in these parameters. Ten Bos taurus crossbred non-lactating beef cows (body condition 4–8 on 9-pt scale; mean = 6.7) were used in the experiment. Dominant follicle removal was performed 5 days before sample collection and antral follicle count. Blood was collected through caudal tail venipuncture in serum collection tubes. Antral follicle count was performed through trans-vaginal ultrasound imaging of ovaries where follicles ≥2 to 3 mm were counted. Serum AMH concentration was measured utilising the Bovine AMH ELISA kit AL-114. Blood sampling and antral follicle count was performed at midmonth during April, May, June, and July. Serum AMH and antral follicle count was analysed by ANOVA. Days with average temperature-humidity index (THI) >75 (mild stress) per month were 0/30, 6/31, 26/30, and 31/31 for April, May, June, and July, respectively. Days with average THI >79 (high stress) per month were 0/30, 0/31, 6/30, and 14/31 for April, May, June, and July, respectively. The total antral follicle count for both ovaries (mean ± SE) was 36.1 ± 4.20, 48.1 ± 7.19, 34.2 ± 4.19, and 46.5 ± 5.85 for April, May, June, and July, respectively. There was no difference (P = 0.1936) in antral follicle count between months. Serum AMH concentration (ng mL−1) was 0.554 ± 0.14, 0.857 ± 0.21, 0.513 ± 0.12, and 0.575 ± 0.13 for April, May, June, and July, respectively. There was no difference (P = 0.3851) in AMH serum concentration between months. The concentration of AMH/follicle (AMH serum concentration/antral follicle count; ng mL−1) was 0.0150 ± 0.0028, 0.0166 ± 0.0018, 0.0152 ± 0.0036, and 0.0123 ± 0.0025 for April, May, June, and July, respectively. There was no difference (P = 0.7385) in the concentration of AMH/follicle between months. Results of the experiment showed that heat stress does not affect antral follicle count, AMH serum concentration, or concentration of AMH/follicle and suggest that granulosa cell function in terms of AMH production is not affected by heat stress during the spring to summer transition.

During oocyte collection, follicular wave phase is unknown, although differences in follicle environment may have dramatic effects on oocyte quality. This project was performed to determine whether oocyte collection during different phases of the follicular wave affects oocyte competence. Oocytes were collected via transvaginal ultrasound guided oocyte aspiration from 18 cows, at 4, 8, and 12 days following dominant follicle removal, representing follicle wave emergence, peak, and atresia, respectively (160, 314, and 273 oocytes, respectively). Once recovered, oocytes were graded and assigned to either being held as immature or matured in vitro for 24 h. Oocytes were then stained in Mitotracker deep red, fixed and stained with an anti-IP3R1 primary antibody and an Alexa Fluor 488-conjugated secondary antibody, before being stained with DAPI, to identify mitochondria, inositol triphosphate receptor 1 (IP3R1), and chromatin respectively. Mitochondria were analysed based on cytoplasmic distribution and classified as peripheral (immature), diffuse, central (mature), or sparse. Expression of IP3R1 was measured as corrected total cell fluorescence in Image J (National Institutes of Health, Bethesda, MD, USA). Staining patterns were analysed using ANOVA. A subset of the matured oocytes was stained with Fluo-3 to measure cytoplasmic calcium levels. These oocytes were then parthenogenetically activated before being imaged again to view changes in calcium levels, and presumptive embryos were cultured for 4 days. Fluo staining was measured as intensity levels (none, slight, moderate, high) and differences in development and stain levels were analysed using the Kruskal-Wallis test. Although mitochondria location was unaffected by collection day, it was significantly affected by maturation status (P = 0.0036). However, oocytes showed incomplete mitochondrial maturation, with mitochondria residing in the diffuse orientation in the majority of oocytes. Expression of IP3R1 appeared to be more sensitive to treatment. Expression significantly increased as meiosis proceeded (P = 0.0081) and there was a significant difference in expression between oocyte collection days (P = 0.0026). The interaction between collection day and maturation status also had a significant effect (P = 0.048), with mature oocytes showing an increase in IP3R1 expression, most notable in those collected on Day 4. Oocyte quality had a notable effect on the ability of oocytes to progress through meiosis (P = 0.054) and on mitochondrial location (P = 0.053), with AB oocytes showing better maturation parameters in both respects. Although the day of collection did not affect embryo development, Fluo stain intensity was an indicator of embryo developmental potential (P = 0.053), with oocytes having decreased potential to develop if the initial calcium levels were moderate to high. Results suggest that oocyte collection during wave emergence yields a slight advantage in oocyte quality. Although IP3R1, necessary for Ca2+ spikes during fertilization, indicates competence, high levels of cytoplasmic Ca2+ at the time of activation appear to be detrimental to embryo development.

Purpose ± Powder bed fusion additive manufacturing (PB F AM) of metal components has attracted much attention, but the inability to quickly and easily ensure quality has limited its industrial use. Since the technology is currently being investigated for critical engineered components and is largely considered uns uitable for high volume production , traditional statistical quality control methods cannot be readily applied . An alternative strategy for quality control is to monitor the build in real time with a variety of sensing methods and, when possible, to correct any defects as they occur. This article reviews the cause of common defects in powder bed additive manufactur ing , briefly surveys process monitoring strategies in the literature, and summarizes recently -developed strategies to monitor part quality during the build process. Design/methodology/approach ± Factors that affect part quality in powder bed additive manuf acturing are categorized as those influenced by machine variables and those affected by other build attributes. Within each category, multiple process monitoring methods are presented. Findings ± A multitude of factors contribute to the overall quality of a part built using PB F AM . Rather than limiting processing to a pre -defined build recipe and assuming complete repeatability , part quality will be ensured by monitoring the process as it occurs and , when possible, altering the process conditions or build plan in real -time . Recent work shows promise in this area and brings us closer to the goal of wide -spread adoption of additive manufacturing technology. Originality/value - This work serves to introduce and define the possible sources of defects and errors in metal -based PB F AM, and surveys sensing and control methods which have recently been investigated to increase overall part quality . E mphasis has been placed on novel developments in the field and their contribution to the understanding of the add itive manufacturing process. Keywords ± additive manufacturing, sensing and control, process quality, defects Article Classification - General review

Fetal disorders of mitochondrial fatty acid oxidation have recently been associated with obstetric complications including pre-eclampsia, Hemolysis, Elevated Liver enzymes, Low Platelets (HELLP) syndrome, placental floor infarct, and Acute Fatty Liver of Pregnancy (AFLP). These diseases occur in about a third of the mothers who are heterozygous for a defect in long chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) enzyme and who bear a fetus homozygous for the defect. The mechanism of this association is not clearly understood. In this study, we provide evidence that the placenta may be the site of production of toxic intermediates of fatty acid metabolism, which accumulate to cause liver damage in the mother. We show that two critical enzymes of long chain fatty acid metabolism, long chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) and short chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD), are active in the normal human placenta. There is an inverse correlation between the enzyme activity of both the enzymes and maternal gestational age during the second and third trimesters. We believe that the demonstration of fatty acid oxidation enzyme activity by the placenta is the first step towards assessing a possible role for fetal/placental fatty acid oxidation defects in the pathogenesis of a subset of pregnancy complications.

While vitrification has become a valuable system used in oocyte and embryo preservation, there is still much to be learned in optimizing this protocol. Both mature and immature oocytes can be vitrified but each presents challenging aspects. Mature oocytes have microfilaments that are not yet developed in immature oocytes, which are fragile and may be disrupted by ice crystal formation during freezing. Further, currently many different cryoprotectants are used in different concentrations, most being combinations of dimethyl sulfoxide (DMSO), glycerol, and ethylene glycol. This study aimed to determine if vitrification solutions composed of ethylene glycol and either dimethyl sulfoxide or glycerol resulted in more-competent post-thaw oocytes, and to determine if maturation stage affected optimal vitrification solution. As validation of the IVF protocol, fresh mature oocytes from a commercial source were fertilized and proportion, with pronuclei formation 48 h post-IVF was recorded. Two experiments evaluated 2 cryoprotectant solutions by analysing post-vitrification and thaw competence of in vitro-fertilized oocytes to form pronuclei. Oocytes in both studies were exposed to 2 sequential vitrification solutions containing 10% DMSO or glycerol, 10% ethylene glycol and 0.5 M sucrose, and then 20% DMSO/glycerol and ethylene glycol and 0.5 M sucrose, before vitrification on cryolocks. In the first study, immature bovine oocytes (n = 200) were vitrified. Following thawing and IVM, they were analysed for pronuclei formation, with 8.49% and 0% fertilization following vitrification in DMSO and glycerol, respectively (P

Although vitrification is becoming increasingly common for oocyte preservation, there has been recent evidence that some cryoprotectants may alter DNA methylation and so result in decreased oocyte developmental competence and abnormal embryonic development post-warming. The objective of this study was to determine the effect of dimethyl sulfoxide (DMSO)- or glycerol-based vitrification protocols on DNA methylation in bovine cumulus-oocyte complexes (COC). The vitrification protocols evaluated included a combination of ethylene glycol (EG) with either DMSO or glycerol (GLY). Cumulus-oocyte complexes were first exposed to equilibration solution (ES) consisting of 7.5% DMSO or GLY and 7.5% EG for 9 min at room temperature (RT) before being transferred to vitrification solution (VS) containing 15% DMSO or GLY, 15% EG, and 0.5 M sucrose. While in VS, 3 to 4 COC were loaded into an open system vitrification device (Cryolock®, BioTech Inc.) and plunged into LN within 1 min. For warming, COC were exposed to dilution solution 1 consisting of 0.5 M sucrose (37°C) for 2.5 min and to dilution solution 2 consisting of 0.25 M sucrose for 2.5 min (RT). Base media for all solution was PBS supplemented with 20% FBS. Cumulus-oocyte complexes (n = 137) were collected from crossbred cows by ovum pick-up on 3 separate days (repetitions), with half of the oocytes each day being matured to metaphase II (MII) stage before vitrification and the other half vitrified as germinal vesicle (GV) COC. The resulting treatments were DMSO GV v. Glycerol GV and DMSO MII v. Glycerol MII. Fresh COC (GV and MII stage, respectively) were utilised as controls. DNA methylation analysis of oocytes was performed through fluorescent staining using a primary antibody anti-5 mC (1:1000) and secondary antibody Alexa Fluor 488 (1:500). Fluorescence intensity of samples was measured using ImageJ software (NIH, Bethesda, MD, USA). Data were analysed through ANOVA with post hoc Tukey’s test. No differences between the vitrification groups or controls were found when analysing DNA methylation of GV stage COC (P = 0.1825). Metaphase II stage COC showed statistical difference between groups (P

Progesterone is a key hormone in reproduction and plays a crucial regulatory role during the oestrous cycle and pregnancy. The effect of progesterone on the developmental competence of bovine oocytes is yet to be fully understood. Molecular tools might provide further information on the role of progesterone on expression of developmentally important genes. The objective of this study was to evaluate the effect of the prolonged exposure of beef cows (days = 32) to different progesterone levels on the expression of genes associated with oocyte developmental competence. Fifteen cows were used in the experiment and were divided in 3 treatment groups (group n = 5). Treatment 1 consisted of no progesterone supplementation, treatment 2 consisted of supplementation by insertion of one controlled internal drug release device (CIDR) per animal, and treatment 3 consisted of supplementation with the insertion of 2 CIDR per animal. All cows were subjected to follicle ablation 4 days before initiation of the experiment. At Day 0, prostaglandin F2α (PGF2α, 25 mg of dinoprost tromethamine, i.m., Lutalyze®) was administered and CIDR devices were inserted in cows designated to treatments 2 and 3. Cumulus-oocyte complexes were collected through ovum pickup where follicles >2 mm were aspirated. Samples evaluated were obtained at Day 0, 8, 16, 24, and 32. The oocyte developmental competence-associated genes METAP2, FOXM1, HMGA1 were analysed in the study. Polyadenylate polymerase was used as normalization gene. Relative gene expression was calculated utilising a comparative CT method (2−ΔΔCT) where data were calculated as fold increase (>1) or decrease ( 0.05). Similarly, no significant differences between treatments was detected in relative expression of METAP2, FOXM1, or HMGA1 across time (P > 0.05). Prolonged exposure of progesterone at different levels does not affect the expression of METAP2, FOXM1, and HMGA1 genes in bovine cumulus-oocyte complexes as indicated by the results of this experiment.