Membrane progesterone receptors mediate progesterone-stimulated glycogenolysis in the bovine uterine epithelium.

In livestock, the amount of glucose needed by the endometrium and embryo increases during early pregnancy. Yet, how glucose concentrations in the endometrium are regulated remains unclear. The bovine uterine epithelium can store glucose as glycogen, and glycogen content decreases in the luteal phase. Our objective was to elucidate the role of progesterone in glycogen breakdown in immortalized bovine uterine epithelial (BUTE) cells. After 48 hours of treatment, progesterone decreased glycogen abundance in BUTE cells (P<0.001) but did not alter glycogen phosphorylase levels. RU486, a nuclear progesterone receptor (nPR) antagonist, did not block progesterone's effect, suggesting that progesterone acted through membrane progesterone receptors (mPRs). RT-PCR confirmed that BUTE cells express all 5 mPRs, and immunohistochemistry showed that the bovine uterine epithelium expresses mPRs in vivo. A mPRα agonist (Org OD 02-0) reduced glycogen abundance in BUTE cells (P<0.001). Progesterone nor Org OD 02-0 affected cAMP concentrations. Progesterone increased phosphorylated AMP-activated protein kinase (pAMPK) levels (P<0.001), indicating that progesterone increases intracellular AMP concentrations. However, AMPK did not mediate the effect of progesterone. AMP allosterically activates glycogen phosphorylase, and D942 (which increases intracellular AMP concentrations) decreased glycogen abundance in BUTE cells. A glycogen phosphorylase inhibitor partially blocked the effect of progesterone (P<0.05). Progesterone and Org OD 02-0 had similar effects in Ishikawa cells (P<0.01), a human cell line that lacks nPRs. In conclusion, progesterone stimulates glycogen breakdown in the uterine epithelium via mPR/AMP signaling. Glucose released from glycogen could support embryonic development or be metabolized by the uterine epithelium.

The glycogenolytic enzyme acid α-glucosidase is expressed in the bovine uterine endometrium.

Progesterone has been shown to stimulate glycogen catabolism in uterine epithelial cells. Acid α-glucosidase (GAA) is an enzyme that breaks down glycogen within lysosomes. We hypothesized that progesterone may stimulate glycogenolysis in the uterine epithelium via GAA. We found that GAA was more highly expressed in the stroma on Day 1 than on Day 11. However, GAA did not appear to differ in the epithelium on Days 1 and 11. Progesterone (0-10 µM) had no effect on the levels of the full-length inactive protein (110 kDa) or the cleaved (active) peptides present inside the lysosome (70 and 76 kDa) in immortalized bovine uterine epithelial (BUTE) cells. Furthermore, the activity of GAA did not differ between the BUTE cells treated with 10 µM progesterone or control. Overall, we confirmed that GAA is present in the cow endometrium and BUTE cells. However, progesterone did not affect protein levels or enzyme activity.

Insertion-Deletion Events Are Depleted in Protein Regions with Predicted Secondary Structure.

A fundamental goal in evolutionary biology and population genetics is to understand how selection shapes the fate of new mutations. Here, we test the null hypothesis that insertion-deletion (indel) events in protein-coding regions occur randomly with respect to secondary structures. We identified indels across 11,444 sequence alignments in mouse, rat, human, chimp, and dog genomes and then quantified their overlap with four different types of secondary structure-alpha helices, beta strands, protein bends, and protein turns-predicted by deep-learning methods of AlphaFold2. Indels overlapped secondary structures 54% as much as expected and were especially underrepresented over beta strands, which tend to form internal, stable regions of proteins. In contrast, indels were enriched by 155% over regions without any predicted secondary structures. These skews were stronger in the rodent lineages compared to the primate lineages, consistent with population genetic theory predicting that natural selection will be more efficient in species with larger effective population sizes. Nonsynonymous substitutions were also less common in regions of protein secondary structure, although not as strongly reduced as in indels. In a complementary analysis of thousands of human genomes, we showed that indels overlapping secondary structure segregated at significantly lower frequency than indels outside of secondary structure. Taken together, our study shows that indels are selected against if they overlap secondary structure, presumably because they disrupt the tertiary structure and function of a protein.

Relationship between donor ejection fraction, left ventricular wall thickness and mortality in heart transplants recipients.

This study explored the impact of donor left ventricular ejection fraction (EF) and left ventricular wall thickness (LVWT) on mortality among heart transplant (HTx) recipients. Utilizing data from the United Network for Organ Sharing (UNOS) registry, adult HTx recipients between 2006-2022 were analyzed. Patients were categorized into four groups based on donor EF(>50 % or ≤50 %) and LVWT(<1.4 cm or ≥1.4 cm). 21,012 patients were included. There were significant differences in baseline characteristics among the groups. Unadjusted mortality was 6.3 %, 6.0 %, 6.0 %, and 2.4 %(p=0.86) at 30-days; 16.2 %, 13.5 %, 16.8 %, and 7.3 %(p=0.08) at 1-year; and 32.2 %, 29.2 %, 35.4 %, and 29.0 %(p=0.18) at 5-years, respectively. In addition, adjusted mortality did not differ across the groups. There were no significant differences in recipient mortality in groups based on donor EF and LVWT. Expanding the donor selection criteria would allow for increase in the donor pool and assist in decreasing the mortality, while on the waitlist for HTx.

Endometrial Glucose Metabolism During Early Pregnancy.

Approximately 50% of human pregnancies humans fail, most before or during implantation. One factor contributing to pregnancy loss is abnormal glucose metabolism in the endometrium. Glucose contributes to preimplantation embryo development, uterine receptivity, and attachment of the embryo. Across multiple species, the epithelium stores glucose as the macromolecule glycogen at estrus. This reserve is mobilized during the preimplantation period. Glucose from circulation or glycogenolysis can be secreted into the uterine lumen for use by the embryo or metabolized via glycolysis, producing ATP for the cell. The resulting pyruvate could be converted to lactate, another important nutrient for the embryo. Fructose is an important nutrient for early embryos, and the epithelium and placenta can convert glucose to fructose via the polyol pathway. The epithelium also uses glucose to glycosylate proteins, which regulates embryo attachment. In some species, decidualization of the stroma is critical to successful implantation. Formation of the decidua requires increased glucose metabolism via the pentose phosphate pathway and glycolysis. After decidualization, the cells switch to aerobic glycolysis to produce ATP. Paradoxically, the decidua also stores large amounts of glucose as glycogen. Too little glucose or an inability to take up glucose impairs embryo development and decidualization. Conversely, too much glucose inhibits these same processes. This likely contributes to the reduced pregnancy rates associated with conditions like obesity and diabetes. Collectively, precise control of glucose metabolism is important for several endometrial processes required to establish a successful pregnancy. The factors regulating these metabolic processes remain poorly understood.

Shared E-Scooter Trajectory Analysis During the COVID-19 Pandemic in Austin, Texas.

By March of 2020, most cities worldwide had enacted stay-at-home public health orders to slow the spread of COVID-19. Restrictions on nonessential travel had extensive impacts across the transportation sector in the short term. This study explores the effects of COVID-19 on shared e-scooters by analyzing route trajectory data in the pre- and during-pandemic periods in Austin, TX, from a single provider. Although total shared e-scooter trips decreased during the pandemic, partially owing to vendors pulling out of the market, this study found average trip length increased, and temporal patterns of this mode did not meaningfully change. A count model of average daily trips by road segment found more trips on segments with sidewalks and bus stops during the pandemic than beforehand. More trips were observed on roads with lower vehicle miles traveled and fewer lanes, which might suggest more cautious travel behavior since there were fewer trips in residential neighborhoods. Stay-at-home orders and vendor e-scooter rebalancing operations inherently influence and can limit trip demand, but the unique trajectory data set and analysis provide cities with information on the road design preferences of vulnerable road users.

Cardiac and noncardiac biomarkers in patients undergoing anthracycline chemotherapy - a prospective analysis.

BACKGROUND: Biomarkers represent a potential tool to identify individuals at risk for anthracycline-induced cardiotoxicity (AICT) prior to symptom onset or left ventricular dysfunction. METHODS: This study examined the levels of cardiac and noncardiac biomarkers before, after the last dose of, and 3-6 months after completion of doxorubicin chemotherapy. Cardiac biomarkers included 5th generation high-sensitivity cardiac troponin T (cTnT), N-terminal pro-brain natriuretic peptide, growth/differentiation factor-15 (GDF-15), and soluble suppression of tumorigenesis-2 (sST2). Noncardiac biomarkers included activated caspase-1 (CASP-1), activated caspase-3, C-reactive protein, tumor necrosis factor-α, myeloperoxidase (MPO), galectin-3, and 8-hydroxy-2'-deoxyguanosine. Echocardiographic data (LVEF and LVGLS) were obtained at pre- and post-chemotherapy. Subanalysis examined interval changes in biomarkers among high (cumulative doxorubicin dose ≥ 250 mg/m2) and low exposure groups. RESULTS: The cardiac biomarkers cTnT, GDF-15, and sST2 and the noncardiac biomarkers CASP-1 and MPO demonstrated significant changes over time. cTnT and GDF-15 levels increased after anthracycline exposure, while CASP-1 and MPO decreased significantly. Subanalysis by cumulative dose did not demonstrate a larger increase in any biomarker in the high-dose group. CONCLUSIONS: The results identify biomarkers with significant interval changes in response to anthracycline therapy. Further research is needed to understand the clinical utility of these novel biomarkers.

Safe streets for some: A review of local active transportation responses across the U.S. during the COVID-19 pandemic.

Introduction & research objectives: The COVID-19 pandemic significantly disrupted daily travel. This paper contrasts 51 US cities' responses, namely street reallocation criteria and messaging related to physical activity (PA) and active transportation (AT) during the early months of the pandemic. This study can be utilized by cities for aiding in the creation of locally responsive policies that acknowledge and remedy a lack of safe active transportation. Methods: A content analysis review was conducted of city orders and documents related to PA or AT for the largest city by population in all 50 US states and the District of Columbia. Authoritative documents issued from each city's public health declaration (ca. March 2020) to September 2020 were reviewed. The study obtained documents from two crowdsourced datasets and municipal websites. Descriptive statistics were used to compare policies and strategies, with a focus on reallocation of street space. Results: A total of 631 documents were coded. Considerable variation existed in city responses to COVID-19 that impacted PA and AT. Most cities' stay-at-home orders explicitly permitted outdoor PA (63%) and many encouraged PA (47%). As the pandemic continued, 23 cities (45%) had pilot programs that reallocated street space for non-motorized road users to recreate and travel. Most cities explicitly mentioned a rationale for the programs (e.g., to provide space for exercise (96%) and to alleviate crowding or provide safe AT routes (57%)). Cities used public feedback to guide placement decisions (35%) and several welcomed public input to adjust initial actions. Geographic equity was a criterion in 35% of programs and 57% considered inadequately sized infrastructure in decision-making. Conclusions: If cities want to emphasize AT and the health of their citizens, safe access to dedicated infrastructure needs to be prioritized. More than half of study cities did not instate new programs within the first 6 months of the pandemic. Cities should study peer responses and innovations to inform and create locally responsive policies that can acknowledge and remedy a lack of safe AT.

Retrospective analysis of the impact of pre-transplant implantable cardioverter-defibrillator status on long-term prognosis in heart transplant patients.

BACKGROUND: Sudden cardiac death (SCD) post-heart transplantation affects 8%-35% of patients; however, the risk profile remains to be completely elucidated. While pre-transplant ICDs are typically removed during transplantation, no information exists to suggest if this pre-transplant risk stratification is also associated with post-transplant outcomes. The objective of this study was to assess the impact of pre-transplant ICD status on long-term prognosis post-heart transplant. METHODS: The United Network for Organ Sharing registry was queried for all adult heart transplant recipients from 2010 to 2018. Patients were categorized as with versus without ICD prior to heart transplantation. Survival was compared using Kaplan-Meier analysis. Proportional hazards regression analysis assessed the impact of ICDs adjusting for clinical and demographic covariates. RESULTS: Of 19 026 patients included, 78.6% (n = 14 960) had received an ICD at time of registration. Patients with an ICD were older [54.9 (±11.6) years vs. 48.6 (±15.3) years, p < .001], less likely to be female [25.7% (n = 3842) vs. 31.2% (n = 1269), p < .001], and more commonly diabetic [29.3% (n = 4376) vs 23.5% (n = 954), p < .001]. Kaplan-Meier analysis showed no difference in unadjusted survival trajectory by ICD status (chi-square = .48, p = .49). Survival was unrelated to ICD status in the multivariable model (HR = .98; 95% CI .90-1.07). CONCLUSIONS: Patients receiving an ICD pre-transplant had a higher prevalence of risk factors for SCD than non-ICD patients, yet ICD status prior to heart transplantation was not associated with a change in long-term prognosis post-heart transplantation.

How (some) mammals lost their hair.

An approach that allows scientists to identify regions of the genome that evolved faster in hairless mammals reveals candidate genetic mechanisms that gave rise to hairlessness.

Cholinergic basal forebrain degeneration due to sleep-disordered breathing exacerbates pathology in a mouse model of Alzheimer's disease.

Although epidemiological studies indicate that sleep-disordered breathing (SDB) such as obstructive sleep apnea is a strong risk factor for the development of Alzheimer's disease (AD), the mechanisms of the risk remain unclear. Here we developed a method of modeling SDB in mice that replicates key features of the human condition: altered breathing during sleep, sleep disruption, moderate hypoxemia, and cognitive impairment. When we induced SDB in a familial AD model, the mice displayed exacerbation of cognitive impairment and the pathological features of AD, including increased levels of amyloid-beta and inflammatory markers, as well as selective degeneration of cholinergic basal forebrain neurons. These pathological features were not induced by chronic hypoxia or sleep disruption alone. Our results also revealed that the cholinergic neurodegeneration was mediated by the accumulation of nuclear hypoxia inducible factor 1 alpha. Furthermore, restoring blood oxygen levels during sleep to prevent hypoxia prevented the pathological changes induced by the SDB. These findings suggest a signaling mechanism whereby SDB induces cholinergic basal forebrain degeneration.

Complex genetics cause and constrain fungal persistence in different parts of the mammalian body.

Determining how genetic polymorphisms enable certain fungi to persist in mammalian hosts can improve understanding of opportunistic fungal pathogenesis, a source of substantial human morbidity and mortality. We examined the genetic basis of fungal persistence in mice using a cross between a clinical isolate and the lab reference strain of the budding yeast Saccharomyces cerevisiae. Employing chromosomally encoded DNA barcodes, we tracked the relative abundances of 822 genotyped, haploid segregants in multiple organs over time and performed linkage mapping of their persistence in hosts. Detected loci showed a mix of general and antagonistically pleiotropic effects across organs. General loci showed similar effects across all organs, while antagonistically pleiotropic loci showed contrasting effects in the brain vs the kidneys, liver, and spleen. Persistence in an organ required both generally beneficial alleles and organ-appropriate pleiotropic alleles. This genetic architecture resulted in many segregants persisting in the brain or in nonbrain organs, but few segregants persisting in all organs. These results show complex combinations of genetic polymorphisms collectively cause and constrain fungal persistence in different parts of the mammalian body.

Effect of estradiol and IGF1 on glycogen synthesis in bovine uterine epithelial cells.

In brief: Glucose is an important nutrient for the endometrium and embryo during pregnancy. This study shows that estradiol (E2)/IGF1 signaling stimulates glycogen synthesis in the uterine epithelium of cows, which could provide glucose when needed. Abstract: Glycogen storage in the uterine epithelium peaks near estrus and is a potential source of glucose for the endometrium and embryos. However, the hormonal regulation of glycogen synthesis in the uterine epithelium is poorly understood. Our objective was to evaluate the effect of E2 and insulin-like growth factor 1 (IGF1) on glycogenesis in immortalized bovine uterine epithelial (BUTE) cells. Treatment of BUTE cells with E2 (0.1-10 nM) did not increase glycogen levels. However, treatment of BUTE cells with IGF1 (50 or 100 ng/mL) resulted in a >2-fold increase in glycogen. To determine if the uterine stroma produced IGF1 in response to E2, bovine uterine fibroblasts were treated with E2, which increased IGF1 levels. Immunohistochemistry showed higher levels of IGF1 in the stroma on day 1 than on day 11, which coincides with higher glycogen levels in the uterine epithelium. Western blots revealed that IGF1 treatment increased the levels of phospho-AKT, phospho-GSKß, hexokinase 1, and glycogen synthase in BUTE cells. Metabolomic (GC-MS) analysis showed that IGF1 increased 3-phosphoglycerate and lactate, potentially indicative of increased flux through glycolysis. We also found higher levels of N-acetyl-glucosamine and protein glycosylation after IGF1 treatment, indicating increased hexosamine biosynthetic pathway activity. In conclusion, IGF1 is produced by uterine fibroblasts due to E2, and IGF1 increases glucose metabolism and glycogenesis in uterine epithelial cells. Glycogen stored in the uterine epithelium due to E2/IGF1 signaling at estrus could provide glucose to the endometrium or be secreted into the uterine lumen as a component of histotroph.

Endometrial glycogen metabolism during early pregnancy in mice.

Glucose is critical during early pregnancy. The uterus can store glucose as glycogen but uterine glycogen metabolism is poorly understood. This study analyzed glycogen storage and localization of glycogen metabolizing enzymes from proestrus until implantation in the murine uterus. Quantification of diastase-labile periodic acid-Schiff (PAS) staining showed glycogen in the glandular epithelium decreased 71.4% at 1.5 days postcoitum (DPC) and 62.13% at DPC 3.5 compared to proestrus. In the luminal epithelium, glycogen was the highest at proestrus, decreased 46.2% at DPC 1.5 and 63.2% at DPC 3.5. Immunostaining showed that before implantation, glycogen metabolizing enzymes were primarily localized to the glandular and luminal epithelium. Stromal glycogen was low from proestrus to DPC 3.5. However, at the DPC 5.5 implantation sites, stromal glycogen levels increased sevenfold. Similarly, artificial decidualization resulted in a fivefold increase in glycogen levels. In both models, decidualization increased expression of glycogen synthase as determine by immunohistochemistry and western blot. In conclusion, glycogen levels decreased in the uterine epithelium before implantation, indicating that it could be used to support preimplantation embryos. Decidualization resulted in a dramatic increase in stromal glycogen levels, suggesting it may have an important, but yet undefined, role in pregnancy.

Versican secreted by the ovary links ovulation and migration in fallopian tube derived serous cancer.

High grade serous ovarian cancers (HGSOC) predominantly arise in the fallopian tube epithelium (FTE) and colonize the ovary first, before further metastasis to the peritoneum. Ovarian cancer risk is directly related to the number of ovulations, suggesting that the ovary may secrete specific factors that act as chemoattractants for fallopian tube derived tumor cells during ovulation. We found that 3D ovarian organ culture produced a secreted factor that enhanced the migration of FTE non-tumorigenic cells as well as cells harboring specific pathway modifications commonly found in high grade serous cancers. Through size fractionation and a small molecule inhibitors screen, the secreted protein was determined to be 50-100kDa in size and acted through the Epidermal Growth Factor Receptor (EGFR). To correlate the candidates with ovulation, the PREDICT organ-on-chip system was optimized to support ovulation in a perfused microfluidic platform. Versican was found in the correct molecular weight range, contained EGF-like domains, and correlated with ovulation in the PREDICT system. Exogenous versican increased migration, invasion, and enhanced adhesion of both murine and human FTE cells to the ovary in an EGFR-dependent manner. The identification of a protein secreted during ovulation that impacts the ability of FTE cells to colonize the ovary provides new insights into the development of strategies for limiting primary ovarian metastasis.

Gonococcal endocarditis with aortic root abscess and severe aortic insufficiency.

Gonorrhea caused by Neisseria gonorrhoeae is a very common sexually transmitted infection in the United States and World-wide (Kirkcaldy et al., 2019) [1]. The population with highest prevalence is young sexually active adults aged 15-24 years. In a majority of cases, the presentation is asymptomatic however can range from vaginitis to pelvic inflammatory disease in women. Symptoms in men can include urethritis, epididymitis and proctitis. Disseminated disease is characterized by arthritis and tenosynovitis. Gonococcal endocarditis is a rare but life-threatening complication of disseminated gonococcal infection (DGI) occurring in 1-2% of patients with DGI (Shetty et al., 2004) [3]. Gonococcal endocarditis has a predisposition for aortic valve involvement with formation of large vegetation and valvular ring abscess. Only a handful of cases have been reported in the literature since the advent of penicillin. This case highlights the importance of timely diagnosis and appropriate surgical evaluation to prevent mortality in young patients that may present with this rare complication.

Ex Vivo Ovarian Culture to Model the Initial Metastasis in Ovarian Cancer.

Being able to accurately model metastasis is an important tool in cancer research. Several in vitro and ex vivo models have been developed to model metastasis from the ovary to the omentum, the most frequent metastatic site after leaving the ovary. However, the recent discovery that high-grade serous ovarian cancer (HGSOC) can originate in the fallopian tube and then metastasize to the ovary has necessitated the development of assays that can quantify the adhesion of tumor cells to the ovary. Here we describe a protocol for accessing the adhesion of fluorescent cells to mouse ovaries. This assay can be used to investigate the role of ovarian function, hormones, and adhesion molecules in metastasis of cancer cells originating in the fallopian tube to the ovary, an important step in the progression of HGSOC.

Endometrial glycogen metabolism on days 1 and 11 of the reproductive cycle in dairy cows.

Embryos need glucose or fructose to remain viable; however, it is not well understood how secretion of these carbohydrates is regulated. This study was conducted to evaluate endometrial glycogen and associated enzymes on Days 1 and 11 of the estrous cycle (Day 0 = behavioral estrus) in cattle. Diastase-liable periodic acid-Schiff (PAS) staining of luminal epithelia decreased 81 % between Days 1 and 11. Similarly, glycogen content of glandular epithelia was 66 % less on Day 11 than Day 1. There was dense PAS staining in the lumen of some glands, and this staining was removed when there was pretreatment with diastase. Based on western blot results, there was no difference in glycogen metabolizing enzymes between Days 1 and 11. Results from conducting immunohistochemistry procedures indicated hexokinase 1 was more abundant in the epithelial cells than stroma, but immunostaining was not different between Day 1 and 11. In contrast, phospho-glycogen synthase was undetectable on Day 1 but was present in glandular epithelia on Day 11. Glycogen synthase was localized to the epithelia, and was in larger abundance on Day 1. The abundance of glycogen phosphorylase was greater in the epithelium than stroma and on Day 11 than 1. Furthermore, glucose-6-phosphatase 3 was more abundant in the epithelium on both Days 1 and 11. In conclusion, in the uterus of cattle glycogen is stored in a reproductive cycle-dependent manner. Glucose released from endometrial glycogen stores could potentially be utilized by the endometrium or secreted into the uterine lumen.