Design, synthesis, and biological evaluation of an allosteric inhibitor of HSET that targets cancer cells with supernumerary centrosomes

Centrosomes associate with spindle poles; thus, the presence of two centrosomes promotes bipolar spindle assembly in normal cells. Cancer cells often contain supernumerary centrosomes, and to avoid multipolar mitosis and cell death, these are clustered into two poles by the microtubule motor protein HSET. We report the discovery of an allosteric inhibitor of HSET, CW069, which we designed using a methodology on an interface of chemistry and biology. Using this approach, we explored millions of compounds in silico and utilized convergent syntheses. Only compound CW069 showed marked activity against HSET in vitro. The inhibitor induced multipolar mitoses only in cells containing supernumerary centrosomes. CW069 therefore constitutes a valuable tool for probing HSET function and, by reducing the growth of cells containing supernumerary centrosomes, paves the way for new cancer therapeutics

Cross-Omics Comparison of Stress Responses in Mesothelial Cells Exposed to Heat- versus Filter-Sterilized Peritoneal Dialysis Fluids

Recent research suggests that cytoprotective responses, such as expression of heat-shock proteins, might be inadequately induced in mesothelial cells by heat-sterilized peritoneal dialysis (PD) fluids. This study compares transcriptome data and multiple protein expression profiles for providing new insight into regulatory mechanisms. Two-dimensional difference gel electrophoresis (2D-DIGE) based proteomics and topic defined gene expression microarray-based transcriptomics techniques were used to evaluate stress responses in human omental peritoneal mesothelial cells in response to heat- or filter-sterilized PD fluids. Data from selected heat-shock proteins were validated by 2D western-blot analysis. Comparison of proteomics and transcriptomics data discriminated differentially regulated protein abundance into groups depending on correlating or noncorrelating transcripts. Inadequate abundance of several heat-shock proteins following exposure to heat-sterilized PD fluids is not reflected on the mRNA level indicating interference beyond transcriptional regulation. For the first time, this study describes evidence for posttranscriptional inadequacy of heat-shock protein expression by heat- sterilized PD fluids as a novel cytotoxic property. Cross-omics technologies introduce a novel way of understanding PDF bioincompatibility and searching for new interventions to reestablish adequate cytoprotective responses

Exited Prostitution Survivor Policy Platform

Survivors of prostitution propose a policy reform platform including three main pillars of priority: criminal justice reforms, fair employment, and standards of care. The sexual exploitation of prostituted individuals has lasting effects which can carry over into many aspects of life. In order to remedy these effects and give survivors the opportunity to live a full and free life, we must use a survivor-centered approach to each of these pillars to create change. First, reform is necessary in the criminal justice system to recognize survivors as victims of crime and not perpetrators, while holding those who exploited them fully responsible. Second, reform is necessary to assist survivors in finding fair employment by offering vocational training, financial counseling, and educational scholarships, as well as offering employment opportunities that utilize survivors’ vast array of skills and interests. Finally, standards of care for survivors exiting prostitution should focus on supporting survivors in our journeys and support short- and long-term resources that empower us. These systemic changes are necessary to recognize survivors as the valuable human beings we are and to support survivors in fulfilling our vast potential

Differences in Maternal and Infant Cord Blood Vitamin D Between Racial/Ethnic Groups

Background: Vitamin D deficiency associated with lower 25-hydroxyvitamin D (25(OH)D) concentration is common among individuals with more melanin pigmentation. Low 25(OH)D levels in pregnant women may be related to increased risk of low birth weight and preterm delivery. Still, few studies have assessed how serum levels of 25(OH)D vary between maternal and infant race/ethnicity. Objective: This study aimed to investigate the relationship between 25(OH)D levels in maternal blood and infant cord blood within certain ethnic groups, prematurity status, and low birth weight. Experimental Design:An IRB-approved study enrolled 86 mother-infant pairs. Maternal blood samples and infant cord blood samples were analyzed for 25(OH)D serum levels. Descriptive statistics and Kruskal-Willis tests comparisons were conducted with the use of IBM SPSS Statistics 28 software to assess the relationship between maternal and cord blood 25(OH)D levels in other race/ethnicity groups, birthweight, and preterm birth. Prematurity was categorized into two groups: premature (weeks) and term (≥37 weeks). Birth weight was categorized into two groups: low birth weight (\u3c 2500 g weeks) and not low birth weight (≥2500 g weeks). A p-value of Results:Median levels of 25(OH)D serum were lower in infant’s cord blood (22.52 ng/mL) than maternal blood (38.06 ng/mL). White participants had significantly higher 25(OH)D levels than African American participants in both maternal blood (40.76 ng/mL vs 27.79, p = Conclusion: Our findings suggest a possible association with lower serum 25-hydroxyvitamin D concentration in darker skin pigmentation, even in a small sample size. These results suggest that prematurity and birth weight should be replicated in larger sample sizes of different Race/Ethnic groups, limiting this finding. Further studies should focus on examining differences with larger and more diverse sample sizes. Such research should include measuring Vitamin D intake in pregnancy and clinical outcomes.https://digitalcommons.unmc.edu/surp2021/1008/thumbnail.jp

Assessing The Impact Of Socioeconomic Status On Maternal And Cord Serum Omega-3 Polyunsaturated Fatty Acid Levels

Background: Omega-3 (n-3) polyunsaturated fatty acids (PUFAs) modulate inflammation throughout the lifespan and are essential in fetal growth and development. Previous studies have demonstrated that individuals with lower socioeconomic status (SES) may be at risk for low intake of n-3 PUFAs; however, no research has compared the concentrations of these nutrients present in maternal and cord serum between markers of SES. Significance of Problem: Individuals with low serum levels of n-3 PUFAs may suffer from unfavorable birth and pregnancy outcomes. Therefore, it is important to identify populations who may have decreased serum levels of these nutrients in order to provide nutritional recommendations to optimize prenatal care. Objective: The purpose of this study is to assess the relationship between markers of SES and levels of n-3 PUFAs in maternal and cord serum in a group of patients delivering at a Midwest Academic Medical Center. Methods: An IRB-approved study enrolled mother-infant pairs (n=55) at the time of delivery for collection of maternal and cord serum samples. n-3 PUFA levels quantified included Eicosapentaenoic acid (EPA), Docosahexaenoic acid (DHA), and total n-3 PUFAs. Markers of SES include private vs public insurance, income ≤150% of the poverty line vs \u3e150%, and college degree earners vs no college degree. Descriptive statistics were run for all variables. The Mann-Whitney U test was used to assess differences in n-3 PUFA levels between SES groups. A p\u3c0.05 was considered statistically significant. Results: Median gestational age at delivery was 39.3 weeks in this cohort. Significantly higher nutrient levels were present in college-educated mothers vs less than college-educated mothers for maternal EPA (9.44 µg/mL vs 5.13 µg/mL, p=0.010), cord EPA (1.88 µg/mL vs 1.40 µg/mL, p=0.011), cord DHA (37.96 µg/mL vs 32.80 µg/mL, p=0.014), and total cord n-3 PUFAs (44.23 µg/mL vs 39.34 µg/mL, p=0.024). Median cord EPA levels were significantly higher in those with private insurance compared to public (1.79 µg/mL, 1.18 µg/mL, p=0.022). Additionally, median cord EPA levels were significantly higher in those \u3e150% the poverty line (1.79 µg/mL, 1.10 µg/mL, p=0.030). No other significant differences were observed between SES groups and n-3 PUFA levels. Conclusion: Our findings suggest that individuals with lower SES may be at risk for lower serum levels of n-3 PUFAs in pregnancy, which could predispose them to adverse birth and pregnancy outcomes. Future studies should focus on replicating these results in a larger, more heterogeneous sample and should consider analyzing additional markers of SES.https://digitalcommons.unmc.edu/chri_forum/1051/thumbnail.jp

Tocopherol Content of Maternal Breast Milk and Impact on Neonatal Growth

Background: Vitamin E is a fat-soluble nutrient consisting of α-, β-, δ-, and γ-tocopherol isoforms that has established effects on neonatal growth in utero. Higher maternal plasma tocopherol concentrations in pregnancy are associated with increased neonatal weight, length, and head circumference percentiles at birth. However, less is known about the impacts of tocopherols on post-natal growth. Since many neonates consume maternal breast milk as their post-natal nutrient source, it is important to understand the tocopherol content of breast milk and associations with neonatal growth. Significance of Problem: Although the tocopherols demonstrate positive associations with neonatal growth in utero, the role of these breast-milk-derived nutrients on post-natal growth is unclear. Enhanced understanding of these relationships can help clinicians and mothers ensure ideal nutrition and growth in their neonates. Hypothesis: We hypothesize there will be positive correlations between maternal breast milk tocopherol concentrations and post-natal growth parameters in neonates. Experimental Design: Breast milk samples were collected from postpartum mothers (N=24) whose neonates were admitted to the neonatal intensive care unit (NICU) and analyzed for α-, δ-, and γ-tocopherol concentrations using high performance liquid chromatography (HPLC). Neonatal anthropometric percentiles at 36 weeks corrected gestational age (CGA) and discharge were collected from the medical record. The Fenton growth chart was used for neonates born \u3c37 weeks CGA and the WHO growth chart for neonates born \u3e37 weeks CGA. Spearman correlations assessed the relationships between breast milk tocopherol concentrations and neonatal growth percentiles. A p-value \u3c0.05 was statistically significant. Results: 70% of neonates were born preterm (median CGA=35.9 weeks). At 36 weeks CGA, median growth percentiles were 59.9 for weight, 68.4 for length, and 60.9 for head circumference. At discharge, median growth percentiles dropped to 33.6, 41.6, and 48.3, respectively. Breast milk concentrations of α- and γ-tocopherol were significantly correlated to increased neonatal length percentile at 36 weeks CGA (both R=0.70, both p=0.016), with γ-tocopherol concentration also correlating with increased weight percentiles at 36 weeks CGA (R=0.62, p=0.033). There were no significant associations between breast milk tocopherol content and discharge growth percentiles. Conclusions: This study demonstrates the potential role of increased maternal breast milk tocopherol content on post-natal growth at 36 weeks CGA for preterm neonates. Limitations of this study include a small sample size and variability among neonates in total breast milk consumption prior to reported growth measures. Future research should assess total breast milk intake in neonates and evaluate neonatal plasma tocopherol concentrations.https://digitalcommons.unmc.edu/chri_forum/1048/thumbnail.jp

Evaluation of Tocopherol Isoforms in Maternal Breast Milk and Their Relationship with Maternal Dietary Intake

Background: Vitamin E is an essential, fat-soluble nutrient with four isoforms: α-, β-, δ-, and γ-tocopherol. These isoforms differentially modulate inflammation and show variable associations with perinatal outcomes, such as preterm delivery and Apgar scores. However, little is known about the role of these isoforms on post-natal outcomes and their presence in maternal breast milk, a neonate’s ideal nutrition source. Significance of Problem: To analyze the role of tocopherols on post-natal growth and inflammation, it is critical to first assess their presence in maternal breast milk. Relating these measures to maternal dietary intake can advance our understanding of breast milk micronutrient composition and provide an avenue for counseling lactating mothers on the importance of maternal nutrition to ensure their neonate’s health. Experimental Design: Breast milk samples were collected from postpartum mothers (N=24) whose infants were admitted to the neonatal intensive care unit (NICU) and analyzed for α-, δ-, and γ-tocopherol concentrations using high-performance liquid chromatography (HPLC). Maternal dietary intake was assessed using the Harvard Food Frequency Questionnaire. Median tocopherol concentrations and isoform proportions were generated for breast milk concentrations and intake values. Tocopherol intake adequacy was defined using the Institute of Medicine’s recommendation of 19mg of α-tocopherol daily. Mann-Whitney U-tests compared median breast milk tocopherol concentrations between intake adequate vs. deficient mothers and assessed for differences in isoform proportions between dietary intake and breast milk samples. A p-value \u3c0.05 was statistically significant. Results: 63.6% of mothers had deficient tocopherol intake (median daily intake=15.3mg α-tocopherol). Median concentrations of α-, δ-, and γ-tocopherol (ug/L) in breast milk samples were 3866.5, 768.1, and 118.6, respectively. There were no significant differences in breast milk tocopherol concentrations between intake adequate vs. deficient mothers. For both dietary intake and breast milk, α-tocopherol had the highest relative proportion (MBM=83%, intake=52%), followed by γ-tocopherol (MBM=14%, intake=39%) and δ-tocopherol (MBM=3% intake=8%). Proportions of δ- and γ-tocopherol were significantly higher in dietary intake compared to maternal breast milk (both p\u3c0.001). Conclusions: This study highlights the prevalence of overall tocopherol intake deficiency and increased proportional consumption of δ- and γ-tocopherol among lactating mothers. It also suggests a mechanism for maintaining breast milk α-tocopherol concentrations despite intake deficiency. Differences in the proportions of tocopherol isoforms between breast milk and intake measures further indicates that proportions of individual tocopherol isoforms in breast milk are influenced by factors other than dietary intake.https://digitalcommons.unmc.edu/chri_forum/1049/thumbnail.jp

Omega-3 and Omega-6 Polyunsaturated Fatty Acid Levels in Placental Tissue and Association With Maternal Dietary Intake

Background: Omega-3 (n-3) and omega-6 (n-6) polyunsaturated fatty acids (PUFAs) are precursors to lipid mediators that modulate inflammation throughout the body. Derivatives of n-3 PUFAs have anti-inflammatory properties and promote growth and neurodevelopment in the fetus. Conversely, n-6 PUFA metabolites exert pro-inflammatory effects and may contribute to adverse pregnancy and birth outcomes. Studies have shown that maternal dietary intakes of n-3 and n-6 PUFAs are associated with their levels in maternal serum, but less is known about the association between PUFA intake and placental levels of these nutrients. A fetus must obtain essential PUFAs from their mother through the placenta, therefore it is important to ascertain how maternal diet may impact placental PUFAs. Understanding this relationship may help inform dietary recommendations in pregnancy. Objective: The purpose of this study is to assess the relationship between dietary intake of n-3 and n-6 PUFAs and their levels in placental tissue. Methods: An IRB-approved study enrolled 29 mother-infant pairs who were delivering at a Midwest Academic Medical Center. Placental tissue samples were obtained at delivery and levels of n-3 and n-6 PUFAs (including Eicosapentaenoic acid (EPA), Docosahexaenoic acid (DHA), Docosapentaenoic acid (DPA), a-Linolenic acid (ALA), Linoleic acid (LA), and Arachidonic acid (AA)) were quantified in each sample. The Harvard Food Frequency Questionnaire (FFQ) was used to assess maternal dietary intake of n-3 and n-6 PUFAs. Descriptive statistics were run for all variables. Spearman correlations were used to assess the relationships between dietary intake and placental PUFA levels. A p\u3c0.05 was considered statistically significant. Results: Median gestational age at delivery was 39.7 weeks in this cohort. Significant positive correlations were observed between maternal EPA intake and levels of placental DHA (R=0.52, p\u3c0.01), total placental n-3 PUFAs (R=0.50, p\u3c0.01), and placental AA (R=0.38, p=0.046). Conversely, maternal EPA intake was negatively correlated with the ratio of placental n-6:n-3 PUFAs (R=-0.45, p=0.02). Maternal DHA intake demonstrated significant positive correlations with placental DHA (R=0.46, p=0.01) and total placental n-3 PUFAs (R=0.43, p=0.02). No significant associations were observed between n-6 PUFA intake and placental levels of n-3 or n-6 PUFAs. Conclusion: Our findings suggest that maternal dietary intakes of EPA and DHA are correlated with n-3 and n-6 PUFA levels in the placenta. This information may help guide optimal dietary guidelines for n-3 PUFAs during pregnancy. A limitation to our study is the small sample size; thus, future studies should focus on replicating these results in a larger sample.https://digitalcommons.unmc.edu/chri_forum/1050/thumbnail.jp

Impact of COVID-19 Infection During Pregnancy on Neonatal Birth Outcomes

Approximately 116 million births have been reported worldwide in the nine months following the start of the COVID-19 pandemic. Currently, the effects of COVID-19 infection during pregnancy on birth outcomes are not fully understood. An IRB-approved study enrolled 115 mothers since March 2020, 5 of whom had a confirmed history of COVID-19 infection during pregnancy. For each COVID-19-infected mother, two mothers of similar age, gestation period, and race who were not infected with COVID-19 during pregnancy were matched 2-to-1 for a case-control analysis. Descriptive statistics were generated, and the Mann-Whitney U test was used to compare continuous variables between the two groups. Fisher’s Exact test was used to evaluate categorical outcomes between the groups. Phttps://digitalcommons.unmc.edu/surp2021/1007/thumbnail.jp