bluuebunny/crossref_metadata_2025 huggingface dataset:
Urinary proteome changes were detected earlier than serum biochemical parameters and histopathology changes in a rat thioacetamide‐induced hepatic fibrosis model
Authors: Zhang Fanshuang , Ni Yanying , Yuan Yuan , Gao Youhe
Published: November 2023

Whereas plasma remain homeostatic, urine tends to accumulate and reflect early changes of the body. This property potentially makes urine a better biomarker source for diseases research. Liver fibrosis which results from depositing extracellular matrix (ECM) components is still a reversible pathological condition whereas cirrhosis as the end‐stage of liver fibrosis is irreversible. Consequently, noninvasive early biomarkers are desperately needed. In this study, thioacetamide (TAA) peritoneo‐injection rat model was used to identify differential urinary proteins in liver fibrosis process by using tandem mass tagging and two‐dimensional liquid chromatography tandem mass spectrometry. A total of 766 urinary proteins were identified, 143 and 118 of which were significantly changed in TAA 1‐week and 3‐week groups respectively, compared with the control group. Multiple reaction monitoring (MRM) targeted proteomics was used for further verification of differentially proteins. A total of 40 urinary proteins were statistically significantly changed (p<0.05) and had fold change more than 2. Those urinary protein changes were earlier than serum biochemical parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total protein and histopathology, which have potential as early biomarkers of liver fibrosis.

Support or Funding Information

This work was supported by the National Key Research and Development Program of China (grant number 2016 YFC 1306300); the National Basic Research Program of China (grant number 2013CB530805); and funds from Beijing Normal University (grant number11100704, 10300‐310421102).
Absolute Quantification of RXRG isoforms
Authors: Skotarczak Teagan , Karnath Erin , Monin Lydia , Shimanovich Yana , Bistulfi Gaia
Published: November 2023

Many genes are transcribed as multiple variants known as isoforms. Isoforms are transcripts from the same locus with either a different transcription start site or alternative splicing. Isoforms might be characterized by different functions and/or stability. Different isoforms of the same gene can vary in length from a few bases to several kilobases. In some cases, there is no region of the transcript specific to only one isoform, making quantification problematic. We compared the isoforms' sequences using bioinformatic tools freely available online. We utilized a one‐step real‐time RT‐PCR protocol, using primers specific to one or more isoforms, which included a T7 promoter and terminator sequences. After confirming amplicon identity by DNA sequencing, we built RNA standard curves. Analysis of the data shows acceptable specificity and efficiency for RXRG3 primers as well as primers picking up all three isoforms. We are currently in the process of assessing primers for RXRG1 and RXRG3.

Support or Funding Information

This work was supported by funding from D'Youville College, Buffalo NY
A Novel Transcription Factor Regulates the Oxidative Stress Response in Caulobacter crescentus
Authors: Maragos Nicholas G. , Mohieddin Nasser , Smolarek Brandon , Bryan Aubrey , Peterson Celeste
Published: November 2023

All aerobically growing organisms have developed mechanisms for coping with oxidative stress. While the oxidative stress response has been well‐studied in some bacteria such as Escherichia coli, it is not as well‐characterized in other bacterial groups such as alphaproteobacteria. Here we examine the role of transcription factors in the oxidative stress response in Caulobacter crescentus. Using viability assays, we find that a deletion in the transcription factor CoxR leads to a strain that is more sensitive to hydrogen peroxide treatment but there are no obvious changes in viability in response to DNA damage agents such mitomycin C, and heat shock. Global gene expression and qRT‐PCR analysis suggested that the CoxR deletion strain has altered gene expression of NADPH cycling enzymes. In summary, CoxR is critical for the oxidative stress response.
In Vitro Activation of Purified Aedes Aegypti Mosquito Early Trypsin (AaET) Wild‐type in Comparison to the Inactive AaET Mutant (Active Site Serine to Alanine)
Authors: Lucero Rachael M. , Rascon Alberto A.
Published: November 2023

The female Aedes aegypti mosquito requires specific midgut proteases to degrade blood meals in a biphasic manner for completion of the gonotrophic cycle. Upon acquisition of multiple blood meals, the female anthrophophilic Aedes aegypti mosquito may become infected with several viruses, such as Dengue, Yellow fever, Chikugunya, and Zika viruses. Once infected, the mosquito can transmit these pathogens to its human hosts, making the mosquito an efficient biological vector. Current vector control strategies have not shown to be efficient, therefore studying and potentially inhibiting the required proteases for blood meal digestion could provide a new vector control strategy. This study is focusing on Aedes aegypti Early Trypsin (AaET), an early phase, trypsin‐like serine protease. This enzyme is one of four most abundant midgut proteases involved in blood meal digestion. The specific activity role of AaET wild‐type (WT) in blood meal digestion has yet to be fully elucidated. The enzyme has been solubly expressed using bacteria as a host, which has led to auto‐catalytic activity. Despite this, zymogen AaET (WT) has been successfully purified and activated using a specific buffer exchange approach that removes dithiothreitol (a reducing reagent that was added in order to inhibit auto degradation during the purification process). Proteolytic activity of zymogen and the active form of AaET (WT) were determined using a chromogenic trypsin substrate, BApNA, to compare to published results. To determine if AaET (WT) is truly autocatalytic, an AaET Mutant (MU) was constructed by mutating the Serine residue in the active site to an Alanine, and was subsequently purified to compare to the wild type form.

Support or Funding Information

SC3 Research Grant from NIH
Funcionalized Gold Nanoparticles as Potential New Treatment for Acute Myeloid Leukemia
Authors: Gaiser Ann‐Kathrin , Hafner Susanne , Schmiech Michael , Syrovets Tatiana , Simmet Thomas
Published: November 2023

Surface functionalization of nanoparticles is often used to target specific cell types or cellular organelles. We have previously found that positively charged nanoparticles manufactured from biologically inert polystyrene are highly cytotoxic towards acute myeloid leukemia cell lines while primary macrophages of healthy donors are resistant.

Acute myeloid leukemia (AML) is a malignancy of the myeloid lineage of the hematopoietic system and, to date, the gold standard treatment combines two chemotherapeutic drugs ‐ cytarabine and daunorubicin. The prognosis and treatment outcome of AML strongly depends on the age of the patients. While patients younger than 65 years show a 5‐year overall survival rate of around 50%, older patients show a very poor 5‐year survival of only 4%. Hence, there is still a need for new agents with higher specificity towards AML cells and lower general cytotoxicity.

In the current study we produced and characterized new amino‐functionalized gold nanoparticles and investigated their potential to target specifically leukemia cells, in vitro and in vivo.

The amino‐functionalized gold nanoparticles (AuNP+) exhibit a positive surface charge, and, like the polystyrene particles, proved to be extremely cytotoxic towards AML cells.

The type of cell death was identified to be apoptosis. Further analysis of the underlying mechanism revealed an early loss of mitochondrial functions, including inhibition of mitochondrial respiration and mitochondrial membrane permeabilization. Consequently, this led to release of cytochrome C into the cytosol and caspase activation.

In contrast to positively charged AuNP+, non‐functionalized (AuNP) or carboxy‐functionalized gold nanoparticles with a negative surface charge (AuNP‐) did not show any cytotoxic effect. This differential toxicity of the gold nanoparticles might be due to higher uptake of positively charged AuNP+ particles by AML cells, as detected by transmission electron microscopy.

The positively charged particles also induced cell death in primary human leukemia cells and reduced their colony‐forming potential, while healthy hematopoietic cells were not affected by treatment with AuNP+.

Overall, we have found so far that our positively charged gold nanoparticles induce apoptosis and exhibit selective cytotoxicity towards AML cells. Further we aim to understand the cause of higher sensitivity of AML cells and to investigate the efficacy of AML treatment by AuNP+ particles in a mouse model engrafted with patient‐derived primary AML cells.

Support or Funding Information

VolkswagenStiftung
Values and Perceptions of the Traditional Gross Anatomy Courses at Tufts University School of Medicine
Authors: Jarrah Andrew
Published: November 2023

Human Gross Anatomy has customarily been taught as a didactic course supplemented with practical laboratory instruction. However, given strong directives from the Liaison Committee on Medical Education and other governing bodies, along with advances in educational technology tools, many medical curricula have forgone this format in favor of group‐based learning modalities such as the flipped classroom and Team‐based problem solving. While studies suggest that collaborative learning and technology assisted teaching may improve student performance, other data suggest that didactic formats are actually the preferred learning mode. Tufts University has maintained a traditional lecture and gross lab format with nearly 140 hours of instruction spread over five months of the first year curriculum. The longitudinal nature of the course has allowed for a paced, deep learning of Anatomy in stark contrast with shorter courses which emphasize broad or conceptual learnings. Our preliminary report, which draws on polling results and conversations with the first year class suggests that Gross Anatomy, and the complementing Head & Neck Anatomy course, are one of the most valued components of the curricula at Tufts, not only for their subject matter, but for the breadth and depth as well. Students found that the hands‐on nature of the lab work not only allowed for detailed learning of anatomical complexities, but it also vested a strong respect for the sensitivity of the profession. The pre‐lab lectures were deemed an appropriate primer for all the dissections throughout the course. The current format was found to allow for a mastery of the material due to the comprehensive nature of the didactics along with practical experience in the gross lab. The continued success of this course signals a place for traditional Anatomy instruction in a landscape which continues to evolve towards alternative learning approaches. While controversies still exist, the traditional didactic has proven its ability to deliver deep‐level learning in a format led by experts. Further studies will be needed to show whether technology or peer based learning tools are able to deliver similar results.

Support or Funding Information

This work was made possible by the American Academy of Anatomists and Tufts University School of Medicine.
Effects of Charged Resveratrol Derivatives on Ca2+ Homeostasis in Human Cancer Cells
Authors: Peterson Joshua Allen , Hastings Jordan Parker , Kenealey Jason Donald
Published: November 2023

Resveratrol (RES), a naturally occurring polyphenol found in grapes, peanuts, and berries, selectively facilitates apoptosis in cancer cells. The apoptotic activity of RES in tumor cells is, in part, due to a large, sustained increase in cytosolic calcium ion concentration ([Ca2+]i). Via live cell fluorescence imaging, RES was previously shown to inhibit the plasma membrane Ca2+‐ATPase (PMCA) as a potential part of the apoptotic mechanism in MDA‐MB‐231 human breast cancer cells. The RES‐mediated modulation of [Ca2+]i in cancer cells may also impact other pumps and channels associated with [Ca2+]i homeostasis. The mechanism of action by which RES induces PMCA inhibition is currently unknown. We recently synthesized several charged RES derivatives that allowed us to more thoroughly study the interaction between RES, PMCA and calcium signaling. The membrane‐impermeable, charged RES derivatives are unlikely to be metabolized into a membrane‐permeable RES derivative due to the lack of membrane‐bound esterases in the cancer cell line that we studied. Utilizing live cell microscopy, we were able to determine the extracellular effects of charged RES derivatives on calcium signaling and PMCA inhibition in cancer cells.

Support or Funding Information

This research was funded by a BYU Life Sciences Startup Grant and a BYU Office of Research and Creative Activities (ORCA) Undergraduate Research Grant.
Use of Novel Drug Carrier System to Selectively Deliver Rolipram to the Liver for the Treatment of Alcoholic Liver Disease
Authors: Rodriguez‐Alvarez Walter , Zhang JingWen , Perez‐Abadia Gustavo , McClain Craig , Barve Shirish , Maldonado Claudio , Gobejishvili Leila
Published: November 2023

Work done with phosphodiesterase (PDE) inhibitors has demonstrated their beneficial effects in experimental liver injury. Our group has shown that upregulation of hepatic PDE4 plays a pathogenic role in the development of alcoholic hepatic steatosis as well as cholestatic liver injury in animal models. Due to the potent anti‐inflammatory action of PDE4 inhibitors, two PDE4‐specific inhibitors have been recently approved by FDA for COPD and psoriatic arthritis, however both drugs have CNS associated side effects. Based on our studies demonstrating that hepatic PDE4 upregulation contributes to liver injury, we aimed to develop a liposomal‐based drug delivery system to target the PDE4 inhibitor (rolipram) to the liver, in order to achieve effective therapeutic levels while limiting its access to the CNS and preventing side effects. We demonstrated that our specially formulated and sized fusogenic lipid vesicles (FLVs) begin to selectively accumulate in the liver 10 min after administration. Moreover, when we used FLVs as carrier for rolipram (FLVs‐rolipram), endotoxin‐inducible PDE4 activity was inhibited in the liver without any effect on brain PDE4 activity. Since endotoxemia and increased PDE4 expression plays a critical role in the development of experimental alcoholic liver disease, we then tested the effect of FLVs‐rolipram in a mouse model of alcoholic hepatitis. C57Bl/6 mice were fed the Lieber‐DeCarli liquid diet containing 5% ethanol for 10 days. On day 11, they were gavaged (binged) with 31.5% (vol/vol) ethanol, which is similar to the binge drinking pattern in many alcoholic hepatitis patients. FLVs‐rolipram was administered 12 h before alcohol gavage. Control mice, on an isocaloric liquid control diet, were gavaged with maltose dextrin instead of alcohol. The mice were sacrificed 6 h after alcohol binge. Assays for liver enzymes, liver caspase 3, TUNEL, chloroacetate esterase (CAE), Oil red O staining were performed to document liver injury, steatosis and inflammation. Our results show that FLVs‐rolipram treatment markedly attenuated alcohol induced lipid accumulation in the liver and decreased plasma endotoxin levels. Moreover, hepatic apoptosis and injury was significantly lower in mice treated with FLVs‐rolipram as indicated by decreased hepatic caspase‐3 activity, TUNEL, CAE staining and AST levels. These results demonstrate that the targeted delivery of rolipram to the liver by using our novel FLV drug delivery system, achieves effective therapeutic levels in the liver, while limiting its access to the CNS and preventing side effects.

Support or Funding Information

The work was supported by following grants from NIH: R43AA021331, R44AA021331, R21AA022189, U01AA021901 and P50AA024337
Targeting the transcriptional kinases CDK12 and CDK13 in breast and ovarian cancer
Authors: Hamman Kristin , Bradley Michael , Marineau Jason , Choi Yoon , Malojcic Goran , Orlando David , Ren Yixuan , Ke Nan , Hu Shanhu , Olson Eric , Fritz Christian , Roberts Christopher
Published: November 2023

CDK12 and CDK13 regulate expression of large transcripts requiring substantial processing to produce mature mRNA. This transcriptional regulation includes coordinated phosphorylation of specific repeats within the C‐terminal domain of RNA polymerase II and association with RNA processing factors (Chila, 2016). RNAi knockdown of CDK12 in cell culture decreases expression of DNA damage response genes, including BRCA1 and ATR, while enhancing sensitivity to DNA damaging agents (Blazek, 2011; Liang, 2015). Recently THZ531, a selective covalent inhibitor of CDK12 and CDK13, was shown to decrease expression of DNA damage response genes in cell culture (Zhang, 2016). Here we present further studies with THZ531 to guide our discovery program toward molecules suitable for clinical development and to explore mechanistic rationales for combining a CDK12/13 inhibitor with PARP inhibitors or DNA damaging agents for difficult‐to‐treat cancers such as high‐grade serous ovarian cancer and triple‐negative breast cancer.

Using THZ531 as a benchmark, we developed assays capable of discriminating sub‐nM inhibitors, including quantifying time‐dependent covalent inhibition and cell‐based CDK occupancy. Since CDK7, like CDK12 and CDK13, contains a cysteine residue proximal to the kinase active site, these approaches are critical to understand covalent inhibitor selectivity. Furthermore, we performed kinome paneling studies to better understand selectivity of this scaffold in support of our ongoing efforts to optimize CDK12/13 potency and selectivity.

To pharmacologically investigate the previously reported effects of CDK12 RNAi, growth inhibition of a panel of ovarian and breast cancer cell lines was assessed following treatment with THZ531 (OVA EC50 = 50–200 nM (n=6); BRCA EC50 <50 nM (n=4)). Expression profiling revealed that THZ531 treatment resulted in different sets of genes being affected than was observed following treatment with inhibitors targeting CDK7, CDK9 or BET‐bromodomain proteins. Additionally, CDK12/13, CDK7, and CDK9 inhibitors were profiled in a broad cell line panel (n>400) to reveal relationships between inhibitor sensitivity, mutation status, gene expression, and potential oncology indications that may be addressed by these different mechanisms. Finally THZ531 was synergistic with both PARP inhibitors and DNA damaging agents in ovarian and breast cancer cell lines. These data highlight cancer indications and combinations that may be particularly amenable to treatment with CDK12/13 inhibitors.

While the pharmacokinetic properties of THZ531 preclude adequate target engagement in tumor tissue at tolerated doses in mouse model systems, our ongoing medicinal chemistry program is progressing to identify and optimize CDK12/13 inhibitors suitable for clinical evaluation.
The UDP‐glucuronosyltransferase (UGT) 2A1308Arg variant isoform: Its role in modulating wild‐type UGT2A1 activity and its association with lung cancer risk
Authors: Sutliff Aimee Kristina , Bushey Ryan , Ashmore Joseph H. , Lazarus Philip
Published: November 2023

The UDP‐glucuronosyltransferase (UGT) gene family of enzymes are important in the detoxification of a variety of endogenous and exogenous compounds including hormones, bile acids, drugs and carcinogens. UGT2A1 is expressed in a variety of aerodigestive tract tissues and exhibits glucuronidation activity against polycyclic aromatic hydrocarbons (PAHs). The polymorphic UGT2A1308Arg isoform, encoded by the UGT2A1*3 allele, was previously demonstrated to exhibit no activity against a panel of tobacco carcinogens including a variety of PAHs. In the current study, the effects of UGT2A1308Arg on wild‐type UGT2A1308Gly activity and its potential role in lung cancer risk were investigated. A significant decrease (p<0.005) in UGT2A1 activity was observed against multiple PAH substrates following the co‐expression of UGT2A1308Gly and UGT2A1308Arg in the HEK293 cell line. Co‐immunoprecipitation experiments indicated that negative regulation of wild‐type UGT2A1 activity was due to protein‐protein interactions between the UGT2A1308Gly and UGT2A1308Arg isomers. In a panel of 117 human lung microsomal specimens, the glucuronidation of 3‐hydroxybenzo[a]pyrene (3‐OH‐BaP) was significantly (p<0.05) decreased in specimens from subjects with UGT2A1 (*1/*3) heterozygous alleles versus homozygous UGT2A1*1. No 3‐OH‐BaP glucuronide formation was observed in specimens from subjects with the UGT2A1 (*3/*3) genotype. In case‐control studies, the UGT2A1308Arg variant was found to be significantly associated with increased risk for non‐small cell lung carcinoma (OR=1.4; 95% CI=1.02–1.90) when comparing UGT2A1 (*1/*1) versus [(*1/*3) + (*3/*3)] subjects, a pattern that was strongest in women (OR=1.7; 95% CI=1.06–2.53). The negative modulation of wild‐type UGT2A1 activity against PAHs by the UGT2A1308Arg variant may provide a novel explanation for the variation in enzyme activity observed between individuals and may have important implications in terms of the role of the UGT2A1308Arg variant on tobacco‐related cancer risk.

Support or Funding Information

National Institutes of Health, National Institutes of Environmental Health Sciences [Grant R01‐ES025460]

Health Sciences and Services Authority of Spokane, WA [Grant WSU002292]
Real time imaging of tri‐molecular protein interactions in live cells by Förster resonance energy transfer (FRET) microscopy
Authors: Kuo Hsiang‐Ling , Chang Nan‐Shan
Published: November 2023

Malignant cancer cells frequently secrete significant amounts of transforming growth factor beta (TGF‐beta), hyaluronan (HA) and hyaluronidases to facilitate metastasizing to target organs. In a non‐canonical signaling, TGF‐beta binds membrane hyaluronidase Hyal‐2 for recruiting tumor suppressors WWOX and Smad4, and the resulting Hyal‐2/WWOX/Smad4 complex is accumulated in the nucleus to enhance SMAD‐promoter dependent transcriptional activity. Yeast two‐hybrid analysis showed that WWOX acts as a bridge to bind both Hyal‐2 and Smad4. When WWOX‐expressing cells were stimulated with high molecular weight HA, an increased formation of endogenous Hyal‐2/WWOX/Smad4 complex occurred rapidly, followed by relocating to the nuclei in 20–40 min. In WWOX‐deficient cells, HA failed to induce Smads relocation to the nucleus. To prove the signaling event, we designed a real time tri‐molecular FRET analysis and revealed that HA induces the signaling pathway from ectopic Smad4 to WWOX and finally to p53, as well as from Smad4 to Hyal‐2 and then to WWOX. An increased binding of the Smad4/Hyal‐2/WWOX complex occurs with time in the nucleus that leads to bubbling cell death. In contrast, HA increases the binding of Smad4/WWOX/p53, which causes membrane blebbing but without cell death. In traumatic brain injury‐induced neuronal death, the Hyal‐2/WWOX complex was accumulated in the apoptotic nuclei of neurons in the rat brains in 24 hr post injury, as determined by immunoelectron microscopy. Together, HA activates the Hyal‐2/WWOX/Smad4 signaling and causes bubbling cell death when the signaling complex is overexpressed.

Support or Funding Information

Ministry of Science and Technology, Taiwan
FUT8 Promotes Breast Cancer Cell Invasiveness through Remodeling of TGF‐β Receptor Core Fucosylation
Authors: Tu Cheng‐Fen , Wu Meng‐Ying , Lin Yuh‐Charn , Yang Ruey‐Bing
Published: November 2023

Core fucosylation catalyzed by FUT8 (addition of fucose in α‐1,6‐linkage to core N‐acetyl glucosamine of N‐glycans) is critical for signaling receptors involved in many physiological and pathological processes, such as cell growth, differentiation, cell adhesion, inflammation, and tumor metastasis. TGF‐β‐induced epithelial‐mesenchymal transition (EMT) regulates the invasion and metastasis of breast tumors. However, whether receptor core fucosylation affects TGF‐β signaling during breast cancer progression remain largely unknown. In this study, microarray expression profiling analysis was used to identify that FUT8 is highly upregulated in the TGF‐β‐induced EMT, and is associated with the migratory and invasive ability of a number breast carcinoma cell lines. Consistent with these findings, FUT8 expression correlated with poor disease‐free survival and overall survival of breast cancer patients. In addition, gain‐ and loss‐of‐function studies demonstrate that FUT8 overexpression stimulates EMT process while FUT8 knockdown suppresses the invasiveness of highly aggressive MDA‐MB‐231 and 4T1 breast carcinoma cells. Furthermore, we show that TGF‐β receptor is a direct target of FUT8 and its core fucosylation markedly facilitates TGF‐β binding and enhances downstream signaling. Importantly, pharmacological inhibition by a small molecule inhibitor 2F‐peracetyl‐fucoseo or genetic knockdown of FUT8 suppresses the invasive ability of highly metastatic breast cancer cells in vitro and impair their lung metastasis in vivo, respectively. Together, our results reveal a feed‐forward mechanism of FUT8‐mediated receptor core fucosylation that promotes TGF‐β signaling and EMT, thus stimulates breast cancer cell invasion and metastasis. We will further evaluate the in vivo pathological importance of FUT8 during breast cancer progression, and investigate the translational implication by targeting FUT8 on inhibiting breast cancer metastasis.
Frequency of Antenatal Care Visit is a Key Modifiable Determinant of Low Birth Weight in Malawi
Authors: Chikakuda Aaron T , Song SuJin , Song Won O.
Published: November 2023

Background/Objectives

Low birth weight (LBW) is an important public health indicator for infant morbidity, mortality, and risk for cardiovascular diseases in the later stages of one's life. A LBW prevalence (12%) in Malawi is on a high side with higher disparities in the central region (15%). With this high prevalence rate in Malawi, little is currently known about determinants of LBW in Malawi. We aimed to examine the determinants of LBW in Malawi using the 2010 Malawi Demographic and Health Survey (MDHS) data.

Methods

The MDHS data are collected every 5 years from nationally representative samples. A total of 9,039 women (15–49 y) who had children aged up to 59 mo were included in the study. Samples with biologically implausible birth weights (<500g or >6,000g) and with no data on antenatal care and supplement use were excluded from the analysis. Data on socio‐demographics, antenatal care visit (ANC), and iron and folic acid (IFA) supplement usage of women were collected using a set of questionnaires. Hemoglobin level of women at the time of survey was measured using the HemoCue. Children's birth weights were obtained from their health records or mothers' recall. Multivariate logistic regression was used to identify determinants of LBW.

Results

Most women (82%) lived in rural areas and had primary or less education level (79%). Teenagers and older women were more likely to have LBW babies. Maternal height, education and income level, number of children, IFA supplement usage, number of ANC visits, timing of the first ANC attendance were significantly associated with LBW (p < 0.05). Frequency and timing of the first ANC visits were positively associated with the number of IFA supplements taken during the pregnancy (p<0.001). Women who had <2 ANC visits had higher odds ratio for LBW (OR=1.42, 95% CI=1.09–1.86) compared to those who visited >5 ANC visits after adjusting for maternal age at birth, region, education, wealth index, and birth order of the child.

Conclusion

The frequency of ANC visits is a key modifiable determinant of LBW in Malawi and is associated with use of the IFA during one's pregnancy. We recommend further investigations on barriers of ANC attendance and IFA supplement usage to improve the risk of LWB in this population.
Influence of Maternal Stress, Number of Children, and Marital Status on Maternal Restrictive Feeding Practices of Preschool‐Age Children
Authors: Smith Chelsea Lynn , Swyden Katheryn , Weedn Ashley , Salvatore Alicia L , Terry Robert , Sisson Susan B.
Published: November 2023

Maternal feeding practices contribute to children's development of healthy eating behaviors. Feeding practices, such as restricting certain foods, may be influenced by concern for child weight and maternal stress. While having multiple children in a household may contribute to higher maternal stress, it may be protective against unhealthy feeding practices. Dual parent households may also decrease maternal stress through shared parental demands.

OBJECTIVE

The purpose of this study was to examine the collective influence of maternal stress, number of children, and marital status on maternal restrictive feeding practices.

METHODS

This cross sectional study used online surveys of mothers of 2‐to‐5‐year‐old children to examine associations between maternal stress (Depression, Anxiety, and Stress Scale), number of children, marital status, and feeding practices (Child Feeding Questionnaire). The Child Feeding Questionnaire also evaluated the feeding factor of concern‐for‐child‐weight. Multivariate linear regression was used to assess the relationship between maternal stress, number of children, marital status, and restrictive feeding practices while adjusting for concern‐for‐child‐weight. Structural equation modeling with confirmatory factor analysis was also used to evaluate the multivariate regression model.

RESULTS

Mothers (n=278) were 73% Caucasian, 70% employed full time, 32.5+5.1 years old and the mean number of children in household was 2.0±0.9 children. In individual models, maternal stress contributed to restrictive feeding practices (β=0.017, SE=0.004, p=0.001). Number of children and marital status were not associated with restrictive feeding practices. In multivariate linear regression and structural equation models, maternal stress and number of children in household individually contributed to higher feeding restriction, while their interaction resulted in lower feeding restriction. Concern for child weight also contributed to more restrictive feeding practices in both models.

CONCLUSIONS

Mothers with more children and higher levels of stress exhibited lower feeding restriction. The number of children itself may not protect against restrictive feeding practices, but may contribute to a collective environment where restrictive feeding practices are practiced less often by the mother.
Novel Allele‐Specific Epigenetic Effects Shape Genetic Architecture in the Brain
Authors: Gregg Chris , Huang Wei‐Chao , Ferris Elliott , Cheng Tong , Stacher‐Horndli Cornelia , Gleason Kelly , Tamminga Carol , Wagner Janice , Boucher Ken , Christian Jan
Published: November 2023

Interactions between genetic and epigenetic effects shape brain function, behavior and the risk for mental illness. Random X‐inactivation and genomic imprinting are epigenetic allelic effects that influence genetic architecture and disease risk; however, less is known about other possible epigenetic allelic effects in vivo in the mouse and primate brain. Here, we devised genomics, in situ hybridization and mouse genetics strategies to uncover diverse allelic effects in the brain that are not caused by imprinting or genetic variation. We found allele‐specific expression (ASE) effects that are developmental stage and cell‐type specific, prevalent during neonatal stages of brain development, and cause mosaics of monoallelic brain cells that differentially express wild‐type and mutant alleles for heterozygous mutations. ASE effects are enriched in specific biological pathways, are significantly conserved between primates and mice, and impact risk genes for mental illness in the macaque and human brain. Our findings have potentially broad implications for mammalian brain genetics.

Support or Funding Information

NIH 1R01MH109577‐01 (C.G.), a Simons Foundation Autism Research Initiative Explorer Award (C.G.), a University of Utah Seed Grant (C.G.), a New York Stem Cell Foundation Robertson‐Neuroscience Award (C.G.), and NIH P40RR021380 (JDW).
Development of an Obesity Risk Score for Mothers of Young Children
Authors: Martin‐Biggers Jennifer , Quick Virginia , Byrd‐Bredbenner Carol
Published: November 2023

Mothers are key family food gatekeepers and role models for their young children; research indicates that maternal weights, behaviors, characteristics, and environments affect their children's health and weight. Thus, this study aimed to develop a maternal obesity risk score to help identify non‐obese mothers at risk. Score development was based on a comprehensive survey comprised of valid, reliable instruments that assessed 120 weight‐related behavioral, psychographic, demographic, and home environment characteristics of 550 mothers of preschool children (72% White, 37% with college degree). The Home Obesogenicity Measure of EnvironmentS (HOMES) survey scale scores of all independent variables were entered into a Pearson correlation matrix; the 54 maternal independent variables that were not highly intercorrelated (r2<.5) were entered into a binomial logistic regression model. Maternal weight status, the dependent variable, was dichotomized into not obese and obese. The 54 independent variables explained 60.5% of the variance of maternal obesity. The final model was carefully examined to ensure no multicollinearity by again running Pearson correlations for all independent variables included in the final regression model. The characteristics significantly associated with increased obesity risk were: African American race, lower education, higher number of children, general health rating, more weight teasing history, more concern about child's overweight risk, more conflict in the home, and more body dissatisfaction. These 8 independent variables predicted 53 percent of maternal risk for obesity. Stepwise and hierarchical linear regression were conducted to confirm results of the binomial logistic regression. Both Stepwise and hierarchical linear regression methods confirmed that independent variables significantly predictive of obesity in the binomial logistic regression were significant predictors of maternal obesity risk using other methods. Data were transformed to account for directionality of the variable in the regression model and a summative obesity risk score (possible range 0–8) was calculated for non‐obese mothers (n=384). Mothers were divided into tertiles based on their obesity risk score. Obesity risk scores ranged from 0 to 7, with an average of 5.15±1.64 SD. A comparison of BMI among obesity risk tertile using ANOVA and post‐hoc tests revealed that all tertile pairs differed significantly, with BMI increasing with risk (Low risk BMI 22.3±2.8 SD; Moderate risk 23.4±3.2 SD; High risk 24.6±3.2 SD). Additionally, mothers in the highest tertile had significantly higher intrapersonal, interpersonal, and home environment obesogenic characteristics. This score may be useful in identifying mothers at increased risk of future obesity.

Support or Funding Information

USDA NIFA #2011‐68001‐30170
Modeling how Prognosis Levels of Kidney Disease in Diabetic African American Males Relate to Factors of Fresh Food Availability and Frequency of Communication with Healthcare Providers
Authors: Hopkins‐Harrington Cameron T. , Ongeri Elimelda Moige , Harrison Scott H.
Published: November 2023

Critical factors relating to health disparities have included the degree to which patients may advocate for their own care within a healthcare setting, and the degree to which other factors promoting health are available within the nearby community. Diabetes is a disease that can lead to serious health complications, and has a strong association with diet. We sought to evaluate the progression of diabetic kidney disease in patients having type 2 diabetes, relative to estimated glomerular filtration rates (eGFRs) corresponding to standardized levels of prognosis (G1 to G5). We evaluated the level of communication of patients with their healthcare provider, and the availability of fresh, unprocessed food within their community. Our study examined aggregate data from eight different healthcare providers across the state of North Carolina to evaluate these factors in 4996 African American males ranging in age from 18 to 64 years old. We evaluated for two factors: (i) sources of fresh, unprocessed food that were within three miles of a healthcare provider; and (ii) frequency of telephone calls between patients and their healthcare facilities. These data showed that the two healthcare provider locations that were generally lacking sources of nearby fresh, unprocessed food within a three‐mile radius also had reduced levels of communication with patients having normal or only mildly decreased eGFR activity (G1 and G2; OR=1.75, p<0.05). Further data are needed to examine for how eGFR activity relates to availability of fresh food, but exploratory data analysis has indicated to us some nonlinear interaction across the two factors. We are therefore planning a national study to better evaluate for this kind of specific effect across a larger number of locations of healthcare providers. Overall, our study has indicated the value of a combined study for investigating the relevance of both geographic and communication factors to how healthcare providers are serving African American males having type 2 diabetes.

Support or Funding Information

This study was supported by the Minority Men's Health Initiative (MMHI) through NIH/NIMHD Center Award #U54MD008621 (CFDA 93.307), sub‐award #HU 140004.
Between Tradition and Health: Comparing Community and Expert Perceptions of Hispanic Caribbean Diets in New York City
Authors: Fuster Melissa , Bank Elina
Published: November 2023

Nutrition and health disparities are prevalent among Hispanics in the US. While dietitians can serve as vehicles to promote dietary improvements in this population, there is a paucity of research concerning how these experts perceive the diets of the diverse communities they serve. In addition, most of the research and interventions addressing Hispanics tend to focus on the Mexican‐American population or tackle Hispanics as a homogeneous group, failing to acknowledge the variety of dietary practices and traditions in this population. Addressing these research gaps, the present study sought to (1) evaluate dietitians' perceptions and knowledge concerning traditional Hispanic Caribbean (HC) diets, and (2) compare these perceptions with how members of the HC community evaluate the healthfulness of their diets. The study involved key informant interviews with dietitians working with the HC community in clinical and community settings (n=15), and interviews with members of the HC community (23 adults self‐identified as Puerto Rican, Cuban or Dominican) in NYC. Verbatim transcripts were analyzed using Atlas.ti. The analysis revealed areas of overlap concerning unhealthful aspects of the HC diet (incl. high fat and carbohydrate consumption, low consumption of fruits and non‐starchy vegetables, and the importance of portion control). Community respondents discussed motivations for dietary change, which were compared with dietitians' notions about barriers and best practices to address nutritional issues in this community. The study also identified potential education needs for future nutrition practitioners working with diverse communities. The results from the study contribute to future clinical and community interventions addressing health disparities affecting Hispanic communities, by providing insight on the dietitians' role and approach when working with a specific segment of the Hispanic population.

Support or Funding Information

Research support provided by the City University of New York PSC‐CUNY Award # 69195–00 47 and the NYU Post‐Doctoral Fellowship for Academic Diversity.
Differential Effect of Vitamin D Supplementation on Natural Killer Cell Activity in Normal and Obese Mice
Authors: Lee Ga Young , Cha Kyeong Sun , Park Chan Yoon , Lee Seung Eun , Pae Mun Kyong , Han Sung Nim
Published: November 2023

Vitamin D has an immunomodulatory effect on both innate and adaptive immune responses. Vitamin D has been reported to induce differentiation of monocytes into macrophages, inhibit maturation and differentiation of dendritic cells, and suppress adaptive immunity. Although there are some in vitro studies which showed that vitamin D suppressed Natural Killer (NK) cell activity and IFN‐γ secretion, little is known about the in vivo effect of vitamin D on NK cell functions. In this study, we investigated the in vivo effect of vitamin D on NK cell functions by measuring NK activity, cytokine secretion, activating receptor expression, and degranulation capacity in mice fed control or high‐fat diets containing different levels of vitamin D. Five week‐old C57BL/6 mice were divided into 6 groups by fat amount (10% or 45% kcal fat, CD or HFD) and vitamin D content (50, 1000, or 10000 IU/kg of diet, DD, DC, or DS), then fed diets for 12 weeks. NK cell activity was assessed using radioisotope 51Cr release assay against YAC‐1 target cells. Splenocyte subpopulation was measured by flow cytometry (FACS) analysis. Intracellular expression of IFN‐γ by NK cell, CD4 T cell, and CD8 T cell and proportions of NK cells and CD8 T cells expressing NKG2D and CD107a were determined by FACS analysis upon stimulation of splenocytes with PMA (50 ng/mL)/ionomycin (0.5 μM) for 4 hours. NK activity was significantly higher in NDDS group than HFDDS group at effector to target (E:T) ratios of 100:1, 50:1, and 12.5:1. NDDS group showed significantly higher NK activity compared with NDDD and NDDC groups at E:T ratios of 100:1 and 50:1, whereas there was no significant difference in NK activity among HFDDD, HFDDC, and HFDDS groups. Splenic CD11b‐ NK cell population was significantly higher in NDDS group than HFDDS group, while splenic CD11b+ NK cell population tended to be higher in NDDS group compared with HFDDS group (p=0.057). Percentage of splenic NKT cell was significantly higher in NDDS group than HFDDS group. There were significant differences in the splenic population of CD4 T cell, CD8 T cell, and B cell between ND and HFD groups (p=0.042, p=0.001, p<0.001). Intracellular expression of IFN‐γ and surface expressions of NKG2D, and CD107a in NK cells were not affected by the different levels of vitamin D. However, there were significantly higher surface expressions of CD107a by CD8 T cells in NDDD group compared with NDDS group. In conclusion, vitamin D supplementation enhanced splenic NK activity in control mice, whereas this effect of vitamin D was not observed in high‐fat diet‐induced obese mice. Alternation in splenic NK cell population might have partially contributed to increased NK activity by vitamin D supplementation.

Support or Funding Information

Supported by the grant from the National Research Foundation (NRF) of Korea (NRF‐2015R1D1A1A01059679).
Pre‐hispanic Mexican diet reduced dysbiosis and inflammation mediated by LPS in diet induced obesity
Authors: Avila‐Nava Azalia , Aguilar‐López Miriam , Sanchez‐Tood Monica , Tovar Armando R , Torres Nimbe
Published: November 2023

Obesity is considered a chronic inflammation state accompanied of dysbiosis in the gut microbiota. The purpose of the present work was to study the effect of a combination of foods based on the prehispanic Mexican diet (PMD) (corn, black beans, nopal, tomato, chia and pumpkin seeds on parameters associated with inflammation and gut microbiota. Rats were fed a high fat diet for 6 months, after that, a group of rats were fed the PMD during 3 months, and the rest continued with the high fat/sucrose diet (HFS). At the end of the protocol, feces were collected and DNA was extracted for 16S ribosomal gene sequencing using the MiSeq plataform and a blood sample was collected to determine biochemical parameters. Our results showed that PMD modified the composition of gut microbiota increasing the abundance of Lactobacillus reuteri and Prevotella copri by 30‐fold, Bifidobacterium pseudolongum by 17‐fold and Akkermansia muciniphila by 1.5‐fold with respect to HFS group. On the other hand, PMD decreased the abundance of Staphilococcus epidermis by 70 fold, Bacteroidetes eggerthii by 29 fold, Escherichia coli by 25 fold and Helicobacter apodemus by 6 fold compared to rats fed HFS. Changes in the gut microbiota with the PMD were accompanied with a reduction the inflammatory response by decreasing serum lipopolysaccharide (LPS) levels by 52%. Our results showed that the presence of different compounds such as soluble and insoluble fiber, polyphenols and w‐3 fatty acids with a prebiotic effect, and vegetable protein in the PMD are able to modify gut microbiota decreasing inflammation mediated by LPS caused by obesity.

Support or Funding Information

Financial support by INCMNSZ.
Alaskan Blueberries and Cranberries Restore Glucose Uptake in a Type 2 Diabetes Model
Authors: Collin Aline L , Dunlap Kriya L , Kuhn Thomas B
Published: November 2023

Insulin increases glucose uptake in skeletal muscle cells and adipose tissue by stimulating the recruitment of the glucose transporter 4 (Glut4) to plasma membranes from a pool of storage vesicles. Type II diabetes (T2D) is characterized by an inability to clear blood glucose in response to insulin due to an impairment of Glut4 recruitment and/or function. A persistent, systemic inflammatory and oxidative stress (IOS) is key to the impairment of insulin‐stimulated glucose uptake. Diets high in saturated fatty acids promote insulin resistance mediated by IOS and a disruption of sphingolipid metabolism. T2D incidence has reached epidemic proportions in the US population and worldwide and is strongly associated with obesity. Although the incidence of obesity in Alaska Natives is higher compared to rest of the US population, incidence of T2D is considerably less, which is largely attributed to their subsistence diet rich in Alaska berry fruits.

The objective of this study is to demonstrate whether botanical extracts of wild berries endemic to Alaska harbor the potency to protect or restore insulin‐stimulated glucose uptake utilizing a cellular model of T2D.

Acute insulin exposure of adipocytes and myotubes stimulates a rapid recruitment of Glut4 (glucose transporter 4) to plasma membranes thereby increasing glucose uptake. Cells under control conditions (0.1 mg/ml BSA) responded to increasing concentrations of insulin (100 nM and 200 nM) with an increase in glucose uptake (100 nM: 1.80 +0.43 %, n=8, *p<0.05 and 200 nM: 2.23 + 0.42 %, n=8, *p<0.01) utilizing 2‐NBD‐glucose (2‐(N‐(7‐Nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)Amino)‐2‐Deoxyglucose, a fluorescent glucose reporter). In sharp contrast, pretreatment of cells with 0.75 mM palmitate (saturated C16 fatty acid adsorbed onto BSA) for 18 h significantly abolished insulin‐stimulated glucose uptake (100 nM insulin in palmitate treated cells compared to 100 nM insulin and control p<0.05). Cells pretreated with palmitate and increasing concentrations of insulin (100 nM and 200 nM) did not show a significant difference (100 nM: 1.59 +0.12 %, n=8, and 200 nM: 2.05 + 0.18 %, n=8).

Conclusively, persistent presence of palmitate impairs insulin‐mediated glucose transport and plasma membrane recruitment of Glut4. Supplementation of adipocytes or skeletal myotubes with Alaskan berry botanicals (crude extracts, total anthocyanins, total proanthocyanidins) prior to addition of palmitate partially restored insulin‐stimulated glucose uptake.

Support or Funding Information

Research was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Numbers UL1GM118991, TL4GM118992, or RL5GM118990.

Research was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103395.