bluuebunny/crossref_metadata_2025 huggingface dataset:
Zinc‐induced upregulation of metallothionein (MT)‐2A is predicted by gene expression of zinc transporters in healthy adults
Authors: Chu Anna , Foster Meika , Ward Sarah , Zaman Kamrul , Hancock Dale , Petocz Peter , Samman Samir
Published: November 2023

Marginal zinc deficiency has been suggested to influence multiple physiological systems due to the extensive involvement of zinc in numerous structural and biochemical functions of the body. Recent advancements in molecular techniques have enabled the use of novel approaches in the study of zinc metabolism and homeostasis, especially within the cell. The usefulness of zinc transporter and metallothionein (MT) gene expressions to detect changes in zinc intake remains unclear. This study aims to determine the effects of zinc supplementation on zinc transporters and metallothioneins (MT) and their relationships with physical activity level. Healthy adults (n = 39) were randomised to a zinc group (20 mg Zn/day; n = 19) or no treatment (n = 20). Blood samples were collected on Days 0, 2, 7, 14 and 21. The IPAQ‐short form was used to assess physical activity level at baseline. RNA was extracted from peripheral blood mononuclear cells and gene expression of zinc transporters and MT were analysed using real‐time PCR. Using repeated measures ANOVA, MT‐2A gene expression was higher in the zinc group (P < 0.05) with a significant increase at Day 2 (40 ± 18%, mean ± SE, P < 0.05). In a multiple regression model exploring the changes in all gene expressions between Days 0 and 21, the change in MT‐2A gene expression was correlated with changes in zinc transporter expressions (r2 = 0.54, P = 0.029) with ZIP1 expression emerging as a univariate predictor (P = 0.003). Total physical activity level was positively correlated with ZIP7 gene expression at baseline (r = 0.36, P = 0.029). The influence of physical activity and zinc supplementation on the expression of ZIP7 and MT‐2A respectively demonstrates the effects of external stimuli and mediators of cellular zinc homeostasis, with potential implications for signalling pathways and related cellular functions.

Support or Funding Information

R&D Human Nutrition Grant, Meat and Livestock Australia
CHRONIC ZINC DEFICIENCY ALTERS CHICK (Gallus gallus) GUT MICROBIOTA STRUCTURE AND FUNCTION
Authors: Tako Elad , Koren Omry
Published: November 2023

INTODUCTION

Zinc (Zn)‐deficiency is a highly prevalent micronutrient insufficiency primarily caused by a lack of adequate dietary‐Zn and/or poorly bioavailable dietary‐Zn. Although the gut is a vital organ for Zn‐utilization, and therapeutic Zn‐administration positively influences gut health, the significance of the gut microbial ecology to the host during Zn‐deficiency has yet to be studied.

Objectives

Characterization of distinct bacterial shifts induced by chronic dietary Zn‐depletion.

METHODS/DESIGN

Cecal samples from Zn‐replete (42μg/g‐dietary‐Zn) and Zn‐deficient (2.5μg/g‐dietary‐Zn) treatment groups (Gallus‐gallus, n=14, 4‐weeks) were harvested for bacterial DNA‐extraction and sequenced using bar‐coding of the 16S‐rRNA gene V4 hypervariable region. The diversity of microbiota was assessed through measures of β‐diversity, α‐diversity, and overall species‐richness. Chao1‐index and observed species‐richness were used to assess α‐diversity. We utilized weighted‐UniFrac distances as a measure of β‐diversity to assess the effect of chronic Zn‐deficiency on between‐individual variation in bacterial community composition.

RESULTS

Zn‐depletion induces significant taxonomic shifts and decreases overall species‐richness and diversity, establishing a microbial profile resembling that of various other pathological states. Metagenomic‐analysis showed that predicted KEGG‐pathways responsible for macro‐and micro‐nutrient uptake are significantly depleted in Zn‐deficiency, and, along with decreases in beneficial short‐chain‐fatty‐acids, such depletions induce notable functional and metabolic alterations which may further preclude an optimal host Zn‐status. We identified four bacterial species; Enterococcus‐sp., Ruminococcuslactaris, Clostridium‐lactatifermentans, and Clostridium‐clostridioforme –positively correlated with zinc‐adequacy, and two bacterial species; Clostridium‐indolis and Unclassified‐S24‐7 – positively correlated with Zn‐insufficiency as candidate microbes which may play a significant role in the utilization of dietary Zn during a prolonged deficiency.

CONCLUSIONS

Our results characterize a unique cecal‐microbiota during Zn‐deficiency, and provide evidence for these perturbations as influencers of the Zn‐deficient phenotype. With additional research, such microbial alterations could be used to designate a Zn‐deficiency profile, which could be used as an additional physiological biomarker to establish and quantify deficiency stage and risk.
Serum Zinc and Hemoglobin Concentrations in Persons with Human Immunodeficiency Virus Infection: The Positive Living with HIV (POLH) Study
Authors: Poudel‐Tandukar Kalpana
Published: November 2023

Background

Human Immunodeficiency Virus (HIV) infection has been frequently associated with zinc deficiency and anemia. Zinc deficiency can cause significant impairment in both adaptive and innate immune responses that may cause systemic inflammation and leading to anemia. However, research on the role of zinc in anemia among HIV‐infected persons is limited. We assessed the association between serum zinc and hemoglobin concentrations in a cohort of HIV‐infected persons.

Methods

A cross‐sectional survey was conducted among 311 HIV‐positive people (177 men and 134 women) aged 20 to 60 years residing in Kathmandu, Nepal. Anthropometric and blood pressure examinations were performed and information on medical history, food habits, physical exercise, alcohol, smoking, and types/duration of anti‐retroviral therapy were collected. Fasting blood samples were taken for immunological, lipids, glucose, inflammation, and micronutrient markers evaluations. Serum zinc concentrations were measured by the atomic absorption method. The relationship between serum zinc and hemoglobin concentrations were assessed using multiple linear regression analysis with adjustment for potential cardiovascular, life‐style, and HIV‐related clinical confounding factors.

Results

The means of serum zinc concentrations in men and women were 75.62 (16.27) μg/dL and 72.63 (10.05) μg/dL, respectively and of hemoglobin concentrations were 12.92 g/dL (2.24) and 11.71 g/dL (2.21), respectively. Serum zinc concentrations were positively associated with hemoglobin concentrations (β = 0.02; P = 0.016) in total participants. The relationship remained significant only in men (β = 0.02; P = 0.006) but not in women (β = −0.007; P = 0.719).

Conclusion

Serum zinc concentrations were positively associated with hemoglobin concentrations in HIV‐infected persons. Further prospective study to confirm the role of zinc in anemia among HIV‐infected persons is warranted.

Support or Funding Information

This study was partially supported by the Grant‐in‐Aid for Young Scientists (B) (22790581), Japan Society for the Promotion of Science, The Ministry of Education, Culture, Sports, Science and Technology, Japan; Waseda University Grants for Special Research Projects, General Grant/Ippan Josei, Japan, (2012A‐101); and by the Grant for Research on Global Health and Medicine (No. 21A‐2) from the National Center for Global Health and Medicine, Japan. The funding agency had no role in the study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication.
Selenium (Se) Homeostasis in Chicken Central Nervous System Involves Selenoprotein Expression from Insufficient to Excess Se
Authors: Li Jinlong , Jiang Xiuqing , Cao Changyu , Zhang Cong , Li Xuenan , Li Wei
Published: November 2023

Selenium (Se) incorporated in selenoproteins as selenocysteine and supports various important cellular and organismal functions. When Se deficient, brain present the first priority grade to get it. It becomes a homeostasis of Se content. We recently reported that O‐Phosphoseryl‐tRNA: selenocysteinyl‐tRNA synthase (SepSecS) plays an important role in Se homeostasis regulation in the avian brain (PloS One 7:e35761 (2012)). To better understand the relationship of Se homeostasis and selenoprotein expression in chicken central nervous system (CNS), day‐old HyLine White chickens were fed a low Se diet (Se‐L, 0.028 mg/g) supplemented with 4 levels of dietary Se (0 to 5.0 mg Se/kg) as Na2SeO3 for 8 wk. Then chickens were dissected for getting the CNS, which included cerebral contex, cerebellum, thalamus, bulbus cinereus and marrow. Se content in CNS was measured. The expression of selenoproteome which have 24 selenoproteins were detected by the qRT‐PCR array. Se homeostasis was observed during changed the content of Se in chicken CNS from insufficient to excess Se, especially in cerebral contex and bulbus cinereus. In cerebellum, the expression of SePP1 had augmented when Se content was abundant. It implied that SePP1 was the main Se transporters. In bulbus cinereus, the selenoprotein had increased obviously as the content of Se was 3.0 mg/kg. Furthermore, at the 5.0 mg/kg Se, there was a special increase about the selenoprotein expression in the marrow. The distribution of Se in chicken CNS is very notable. The expression of selenoproteins has a hierarch while changing Se content, and Selt, Selk, Selw, Selu, Gpx3, Gpx4, Sepp1, Selo, Sel15, Seln, Gpx2 and Sepp2 take more necessary function in the chicken CNS. In summary, hierarchy of regulated selenoprotein expression makes an important role of CNS Se metabolism and transport in avian.

Support or Funding Information

China New Century Excellent Talents in University (NECT‐1207‐02), China Program for New Century Excellent Talents In Heilongjiang Provincial University (1252‐NCET‐009), Doctor Initial Funding of Northeast Agricultural University (2012RBC52), Research Special funds for Scientific Innovation Talents of Haerbin (2012RFQXN006) and Science Foundation of the Education Department of Heilongjiang Province (12511031).
High Household Food Insecurity Impacts Dietary Patterns in Rural and Urban American Indian Families with Young Children
Authors: Tomayko Emily , Mosso Kathryn , Cronin Kate , Prince Ronald , Adams Alexandra
Published: November 2023

Objectives

Food insecurity (FI) disproportionately affects American Indian (AI) families, but the relationship between FI and diet in these households is unclear. We sought to characterize FI in rural and urban AI households in 5 diverse communities and its association with dietary intake in both adults and young children.

Methods

In total, 438 adult‐child (2–5 years) dyads in 5 AI communities nationwide participated as part of the Healthy Children, Strong Families obesity prevention trial. Demographic factors were collected, FI was determined using 2 validated items from the United States Department of Agriculture 18‐item Household Food Security Survey, and dietary intake was assessed using food frequency questionnaires. Prevalence of FI was determined, and child and adult diets were compared among food secure and insecure participants using t‐tests. Predictors of FI were examined using logistic regression.

Results

A very high prevalence of FI was determined (61%) and different significantly between urban and rural communities (79.5 and 45%, respectively). FI adults had lower intake of vegetables (P=0.031) and higher intakes of fruit juice (P=0.001), other sugar‐sweetened beverages (P=0.015), and fried potatoes (P<0.001) than food secure adults. FI children had higher intakes of fried potatoes (P=0.033), soda (P=0.010), and sports drinks (P=0.049). Significant predictors of FI included ethnicity (higher for AI), lower educational level, single adult households, WIC participation, and urban settings. Predictors for rural households (increased for single adult household, low educational attainment, working outside the home) differed from urban households (increased with shorter distance traveled to grocery shop, higher number of children in the home) when separate models were run.

Conclusions

This is the first study to examine FI and diet in AI adults and young children concurrently. We identified an alarmingly high overall prevalence of FI and demonstrated dietary intake of adults and children was less optimal in FI households. Interventions for underserved families and communities are urgently needed, with particular attention given to barriers that may be unique to urban families (e.g., absence of a close‐knit community, lack of food sharing practices). Development of local/tribal programs and strong policies to address community FI, food sovereignty, and food access are recommended to supplement and support federal‐level strategies.

Support or Funding Information

This project was funded by the National Institutes of Health, National Heart, Lung, and Blood Institute [grant number 1RO1HL114912] to AA. ET was supported through a National Institutes of Health T32 training grant to the University of Wisconsin, Department of Nutritional Sciences [5T32DK007665]. KM was supported through the Shapiro Summer Research Program through the University of Wisconsin School of Medicine and Public Health.
Individual and Structural Environmental Influences on Utilization of Iron and Folic Acid Supplementation among Pregnant Women in Harare, Zimbabwe
Authors: Tinago Chiwoneso B. , Ingram Lucy Annang , Blake Christine E. , Frongillo Edward A.
Published: November 2023

Objective

The Ministry of Health and Child Care of Zimbabwe established a national policy for the prevention of maternal micronutrient deficiencies that centers on pregnant women receiving daily iron and folic acid (IFA) supplements at their first antenatal care visit and throughout pregnancy. Despite these efforts, utilization of IFA supplementation during pregnancy in Zimbabwe is low and no qualitative data existed to explain these low utilization rates. This study aimed to bridge this knowledge gap by investigating the experiences with and knowledge about IFA supplementation of pregnant women and healthcare workers in Harare, Zimbabwe, the health‐service and social environment in which IFA supplementation occurs, and how these influence utilization of IFA supplementation.

Methods

Semi‐structured, in‐depth, qualitative interviews were conducted in both Shona and English with pregnant women (n=24) and healthcare workers (n=14) providing direct antenatal care services to pregnant women in two clinics located in two low‐income high‐density suburbs in Harare. Data were analyzed thematically using NVivo 10 software.

Results

Influences on utilization from the perspectives of pregnant women and healthcare workers were at both the individual and structural environmental levels. Reasons for low utilization of IFA supplementation were side effects, misconceptions about IFA supplementation, limited access to nutrition information, delayed entry or non‐uptake of antenatal care services, and social norms of pregnant women for micronutrient supplementation. Utilization was enhanced by knowledge of risks and benefits of supplementation, fear of negative health complications with non‐utilization, family support, and healthcare‐provider recommendation for micronutrient supplementation.

Conclusions

Future approaches to improve maternal IFA supplementation utilization in Zimbabwe should address individual and structural environmental factors that begin before pregnancy, continue throughout pregnancy, and reach pregnant women who are currently not seeking antenatal care services. Study findings will be used to inform approaches to strengthen micronutrient supplementation utilization in order to improve the micronutrient status of pregnant women, decrease the micronutrient risk factors associated with maternal mortality, and improve overall maternal and child health in Zimbabwe.

Support or Funding Information

This study was supported by a SPARC Graduate Fellowship from the Office of the Vice President for Research at the University of South Carolina.
Physical Activity and Work Productivity in Response to Iron and Iodine Double‐Fortified Salt Trial in Indian Tea Estate Workers
Authors: Blakstad Mia M. , Nevins Julie E.H. , Venkatramanan Sudha , Przybyszewski Eric M. , Haas Jere D.
Published: November 2023

Introduction

Iron deficiency is the most common micronutrient deficiency in the world, with prevalence estimates for women in low‐income countries ranging between 15 and 50%. Iron is essential for oxygen transport through hemoglobin (Hb), and iron deficiency can lead to decreased physical work capacity. The relationship between iron and work capacity suggests that iron deficient individuals could experience reduced work productivity. We used accelerometry data to determine whether improvements in iron status from an intervention of salt double‐fortified with iron and iodine (DFS) increased the physical activity (PA) and efficiency of work performed by female tea pluckers.

Population

Participants were 212 experienced tea pluckers between 18 and 55 years from a large tea estate in Darjeeling District of northern West Bengal, India.

Methods

Two groups were randomly assigned to receive either salt fortified with potassium iodate, or iodized salt that was additionally fortified with microencapsulated ferrous fumarate (3.3 mg/g salt). Over the 10‐month intervention, women received salt only from the study. All measurements were taken at both baseline and end line. Participants self‐reported ethnicity as Adivasi or Nepali. Weight, height, mid‐upper arm circumference (MUAC) and age were measured by trained research staff. Venous blood was analyzed for Hb, serum ferritin (sFt), and soluble transferrin receptor. Physical activity levels were assessed with ActiGraph GT3x tri‐axial accelerometers worn on the hip daily for six days, except when sleeping or bathing. Energy expenditure during the work day was estimated from heart rate, and productivity was measured as the amount of tea plucked (kg) from morning and afternoon weighing.

Results

Mean (SD) change in minutes from baseline to endline spent at sedentary (<1 MET), light (1–3 MET), and moderate‐to‐rigorous (>3 MET) physical activity during plucking were 32.7 (160.8), 5.6 (75.5), and 6.7 (44.2) respectively. Neither the intervention, change in iron status, energy expenditure, energetic efficiency, nor amount of tea plucked were consistent predictors of PA, even after controlling for potential confounders.

Conclusion

We found that, contradictory to previous findings, improvements in iron status did not improve physical performance or worker productivity in these Indian tea estate workers. The results should be interpreted in light of the very different tea plucking conditions observed during the study period. Our results may highlight the need for measures of work productivity that are more sensitive to changes in iron status.

Support or Funding Information

Funded by the Micronutrient Initiative (MI) of Canada
Early Weight Gain, Linear Growth, and Mid‐childhood Blood Pressure
Authors: Perng Wei , Rifas‐Shiman Sheryl L , Kramer Michael S , Haugaard Line K , Oken Emily , Gillman Matthew W. , Belfort Mandy B.
Published: November 2023

Objective

To investigate relations of change in body mass index (BMI) and length/height during 4 early life age intervals (birth to 6mo, 6mo to 1y, 1 to 2y, and 2 to 3y) with mid‐childhood systolic blood pressure (SBP), and to evaluate for effect modification by birth size (birthweight‐for‐gestational age z‐score).

Methods

We studied 957 participants in Project Viva, an ongoing U.S. pre‐birth cohort. Using multivariable linear regression, we examined the associations of weight gain (change in body mass index [BMI] z‐score) change and linear growth (change in length/height z‐score) during each time period with mid‐childhood SBP, adjusting for maternal education, age, pre‐pregnancy BMI, and smoking; and child's age, sex, race/ethnicity, continuous birthweight‐for‐gestational age z‐score, and growth in preceding periods. We examined interaction of these growth measures with birth size according to small (<10th percentile of sex‐ and gestational‐age specific birthweight), appropriate (10th to <90th percentile), and large‐for‐gestational age (≥90th percentile).

Results

Children were 7.9 ± 0.8 years at time of SBP assessment; 50.3% were girls and most were white (65%). Mean ± SD SBP was 94.4 ± 8.8 mmHg and DBP was 54.3 ± 5.7 mmHg. Each additional z‐score gain in BMI from birth to 6mo and 2 to 3y was associated with 0.81 (95% CI: 0.15, 1.46) and 1.61 (0.33, 2.89) mmHg higher systolic SBP, respectively. Gain in length from 1 to 2y was related to higher SBP among boys (2.24 [0.89, 3.60] mmHg per z‐score) but not girls (−0.18 [−1.60, 1.25] mmHg per z‐score). There was no evidence of effect modification by birth size.

Conclusions

Faster gain in BMI during the first 6 postnatal months and in the preschool years may lead to higher SBP in mid‐childhood, regardless of size at birth. Strategies to reduce accrual of excess adiposity during early life may bring long‐term benefits for cardiovascular health

Support or Funding Information

This work was supported by the US National Institutes of Health (K24 HD069408, R37 HD 034568, P30 DK092924).
JAM‐C deficiency primes endothelial cells for a pro‐inflammatory state
Authors: Reglero Natalia , Beal Robert , Cabrera Claudia , Pickworth Catherine , Golding Matthew , Whiteford James , Nightingale Tom , Imhof Beat , Barnes Michael R , Nourshargh Sussan
Published: November 2023

Endothelial cells (ECs) line the inner surface of all vascular walls and as such form a vital barrier between blood and interstitial tissues. Key to this function is the ability of ECs to form cell‐cell junctions that support the formation of a confluent EC monolayer, providing an effective gate‐keeper for the regulated vascular crossing of blood borne proteins and circulating immune cells. Junctional adhesion molecule‐C (JAM‐C) is a component of EC tight junctions and plays a key role in controlling numerous EC vascular and immune functions. Of note, previous work from our laboratory has shown that certain inflammatory conditions (e.g. ischemia‐reperfusion injury) can elicit reduced expression of EC JAM‐C, a phenomenon that can lead to disrupted neutrophil transendothelial cell migration (TEM) dynamics such as neutrophil reverse TEM (rTEM) (Woodfin et al., Nature Immunol., 2011; Colom et al., Immunity, 2015). To gain a better understanding of the role of JAM‐C in EC phenotype and functions, we conducted an unbiased gene expression analysis of ECs subjected to control or JAM‐C siRNA. Briefly, HUVECs were transfected with control or JAM‐C specific siRNA by nucleofection, with the latter achieving a consistently high (~90%) knockdown (KD) efficiency. Of note, JAM‐C (JAM3) KD did not affect the expression of other key EC junctional markers such as VE‐Cadherin or JAM‐A. The study revealed that JAM‐C KD HUVECs exhibit a pro‐inflammatory gene expression profile under basal conditions. Several genes implicated in leukocyte migration, such as the chemokine IL8 (p=6×10−5 fold change (FC) = 5.1) the adhesion molecules VCAM‐1 (p=9.4×10−6 FC= 16.7) and ICAM‐1 (p=0.0014; FC= 3.7), were significantly upregulated in JAM‐C KD cells. Furthermore, pathway analysis of the gene array data by Ingenuity (IPA) software indicated upregulation of numerous gene clusters associated with cell functions such as cellular homeostasis, leukocyte trafficking and several inflammatory signalling pathways. Regulation of key genes (e.g. IL8, VCAM‐1) was further validated by qPCR and at protein level. On‐going works are addressing the functional implications of these findings. Collectively, the present results have identified the crucial role of JAM‐C in maintaining ECs in a quiescent state, indicating that loss of EC JAM‐C could prime ECs for an enhanced and/or potentially aberrant inflammatory response. These findings provide novel insights into the consequences of reduced expression of JAM‐C under pathological conditions, such as that noted in response to ischemia‐reperfusion injury.

Support or Funding Information

This work was supported by generous funds from the The Wellcome Trust and Marie Curie Actions.
Cell‐type specific mechanisms regulate rhythmic leukocyte migration to tissues
Authors: He Wenyan , Kraus Kerstin , Druzd David , de Juan Alba , Ince Louise , Chen Chien‐Sin , Scheiermann Christoph
Published: November 2023

Leukocyte recruitment from blood to tissues is a critical element of the immune system. However, the dynamics of leukocyte migration over the course of the day are unclear. Specifically, which leukocyte subtypes are being recruited to which specific tissue at what time during the daily cycle is unknown. Here, we screened the main leukocyte subpopulations in murine blood (neutrophils, B cells, CD4 T cells, CD8 T cells, natural killer (NK) cells, NKT cells, eosinophils, inflammatory as well non‐inflammatory monocytes) for oscillations in their migration behaviour. We examined surface protein expression levels of 28 adhesion molecules and chemokine receptors on these leukocyte subtypes in addition to 8 adhesion molecules expressed by endothelial cells in vascular beds of multiple organs using quantitative flow cytometry and imaging‐based approaches. Our data indicate the existence of a rhythmic leukocyte‐and tissue‐specific molecular signature in the expression of pro‐migratory factors. To test the relevance of these oscillations, we performed homing assays to investigate the recruitment capability of individual leukocyte subsets to tissues at specific times. These data revealed strong, >2‐fold time‐of‐day differences in the capacity of cells to emigrate out from blood. In addition, it exposed leukocyte‐distinct rhythms in their homing patterns to tissues, with the microenvironment and leukocyte‐autonomous oscillations co‐contributing in the recruitment process. Specifically, using B‐cell and T‐cell‐specific knockouts of the circadian gene Bmal1 demonstrated altered expression levels of specific oscillatory molecules and leukocyte numbers in tissues, which suggests that rhythmicity in these factors may be directly regulated by the circadian clock. Taken together, our data show discrete tissue and leukocyte mechanisms that control a broad and robustly rhythmic leukocyte recruitment process to organs.

Support or Funding Information

Funded by the European Research Council (CIRCODE) and the German Research Foundation(EmmyNoether SCHE1645/2‐1 and SFB914 project B09).
OxLDL Exerts A Biphasic Effect on Endothelial Function: A Role for Oxidised Lipids in Angiogenesis and Inflammation
Authors: Olding Michael , Ardern‐Jones Michael R , Healy Eugene , Millar Timothy M
Published: November 2023

Oxidised lipids are known to promote inflammation and are implicated in the pathophysiology of psoriasis, promoting cardiovascular disease. The deleterious effects of oxidised low‐density lipoprotein (oxLDL) are potentially mediated through vascular cell activation.

We therefore tested the hypothesis that serum lipids, LDL and its oxidation products can modify endothelial cell function. The effects of serum lipids and oxLDL on primary human endothelial cells were assessed in assays for migration, angiogenesis, leukocyte recruitment and vascular cell surface receptor expression.

Our results showed that at relatively high concentrations oxLDL inhibited angiogenesis and endothelial migration. However, lower concentrations of oxLDL stimulated endothelial migration. In contrast, oxLDL caused increased leukocyte recruitment to the endothelium under shear stress only at higher concentration. Native LDL was shown to have no effect at equivalent concentrations in these assays. Serum lipids were also shown to regulate endothelial function as their removal from cultures inhibited endothelial migration and angiogenesis. However, removal of serum lipids did not impair the ability of cytokine‐stimulated endothelial cells to recruit flowing leukocytes. Analysis of known oxLDL receptor‐1 (LOX‐1) and scavenger receptor B (CD36) showed minimal expression in these cells. A potential mechanism through which oxLDL mediates its activity involves the stimulation of sphingosine metabolism. Initial studies have revealed sphingosine‐1‐phosphate receptor (S1PR1) expression in endothelial cells and is currently under investigation for oxLDL mediated effects.

In conclusion, these data suggest an important role for circulating lipids and lipoproteins in the regulation of endothelial function, which may have implications for psoriatic pathology and cardiovascular disease.

Support or Funding Information

Funded by The Psoriasis Association, UK
Dysfunctional Mechanosensing of Extracellular Matrix in Thoracic Aortic Disease
Authors: Humphrey Jay D
Published: November 2023

The ultimate fate of a compromised thoracic aorta – namely, progressive enlargement and acute dissection or rupture – depends upon the hemodynamic loads and mechanical properties of the wall. We have only limited understanding, however, of the mechanobiological processes that render the thoracic aorta vulnerable to such mechanical failures. Homeostasis requires the intramural cells to sense their local chemo‐mechanical environment and to establish, maintain, remodel, or repair the extracellular matrix to provide suitable compliance and yet sufficient strength. This sensing necessitates a mechanically competent extracellular matrix that can also sequester or present chemo‐ligands to the cells and functional membrane receptors that connect to intact actomyosin filaments of the cytoskeleton and regulate intracellular signaling. Diverse mutant genes associated with these thoracic aortic diseases affect extracellular matrix constituents, membrane receptors, or contractile proteins and associated signaling molecules and poorly controlled hypertension is the major risk factor for the disease. This constellation of mutant genes coupled with the fact that increased biomechanical forces drive the disease suggest that these thoracic aortic conditions result, in part, from a dysfunctional mechanosensing by the intramural cells, which in turn compromises the mechanoregulation of extracellular matrix that is needed to ensure sufficient structural integrity of the wall.

Support or Funding Information

NIH U01 HL116323
Systems Mechanobiology and Multi‐Scale Modeling of Ventricular Hypertrophy and Failure
Authors: McCulloch Andrew D , Buchholz Kyle , Tan Philip , Dewan Sukriti , Saucerman Jeffrey , Omens Jeff H , Nigam Vishal
Published: November 2023

Hypertrophic heart disease is a multi‐factorial, multi‐scale problem in which altered whole body hemodynamics and neurohormonal factors lead to a complex growth and remodeling response in the heart. The cells and extracellular matrix in the myocardium initially compensate to the abnormal environmental factors with altered gene expression, changes in contractile and mechanoenergetic function, cell and tissue structural remodeling, in turn further affecting circulatory dynamics and often leading to decompensation and failure.

We have developed organ scale models of biomechanics growth and remodeling that suggest that a growth law in which myocytes respond differentially to transverse and longitudinal strains can explain many of the differences between ventricular hypertrophy associated with normal postnatal growth, aortic stenosis, mitral valve regurgitation and asymmetric electrical activation [1–3]. In‐vitro studies in micropatterned neonatal mouse ventricular myocytes subjected to anisotropic biaxial stretch show around differences in transcriptional responses when the major axis of stretch is longitudinal compared with transverse to the myofilament axis. Using transcriptomes measured by RNA‐seq in these preparations, we modified a network model of hypertrophic cell signaling [4] to include transverse and longitudinal stretch and transcriptional regulation of gene expression. The results revealed specific differences in cell signaling activated by transverse vs. longitudinal stretch with longitudinal stretch activating more pathways and regulators than transverse strain.

Support or Funding Information

Supported by NIH grants 8 P41 GM103426‐19 (Amaro), P50 GM094503‐01A1 (Beard), 1R01HL105242‐01 (McCulloch/Bers) and 1R01HL121754‐01 (McCulloch/Omens)
Integrating Light‐Sheet and Hemodynamic Forces to Elucidate Cardiac Development, Injury and Repair
Authors: Hsiai Tzung
Published: November 2023

Hemodynamic forces such as shear stress are intimately linked with cardiac morphogenesis. Mutations in Notch signaling components result in congenital heart defects in humans and mice. Hemodynamic shear stress induces vascular endothelial Notch signaling, and Notch pathway mediates differentiation and proliferation of trabecular myocytes. During heart development, the myocardium differentiates into two layers, an outer compact zone and an inner trabeculated zone. The trabeculae form a network of branching outgrowths from the ventricular wall, and both trabeculation and compaction are essential for normal cardiac contractile function. However, the mechanotransduction mechanisms underlying endocardial shear stress (ESS) and the initiation of trabeculation remain elusive. Our multi‐disciplinary approach has demonstrated that peristaltic contraction of the embryonic heart tube produces time‐varying shear stress (¶t/¶t) and pressure gradients (ÑP) across the atrioventricular (AV) canal in a zebrafish model of cardiac development. The advent of zebrafish genetic system has enabled the application of fli1 promoter to drive expression of enhanced green fluorescent protein (EGFP) in all vasculature throughout embryogenesis (Tg(fli1a:EGFP)y1); thereby, allowing for 3‐D visualization of the moving boundary conditions (2D + time) for computational fluid dynamics (CFD) simulation. However, 4‐D (3‐D + time) imaging to recapitulate the endocardium throughout the cardiac cycle requires fast tissue scanning and deep axial penetration. For these reasons, we have developed fluorescent Super‐Resolution Light‐Sheet Microscopy (SRLSM) to capture the 4‐D endocardial trabecular network in response to genetic manipulation of blood viscosity and Notch signaling pathway. Our multi‐disciplinary team aims to establish a fundamental direction at the interface of hemodynamic forces and cardiac development by introduction of a novel 4‐D SRLSM technique to elucidate ESS‐mediated Notch signaling with pathophysiological significance to cardiac development, injury, and repair.

Support or Funding Information

NIH R01HL118650 NIH R01HL083015 NIH R01HL111437 NIH R01HL129727
No Longer a Novelty&[hellip]. Formalizing Ultrasound Based Activities in Gross Anatomy through Objective Structural Practical Examination
Authors: Blankers Tony Robert , Lachman Nirusha , Bhagra Anjali , Pawlina Wojciech
Published: November 2023

Use of ultrasound as a diagnostic tool is fast becoming an expectation in clinical practice. With formal outcomes now being included as competencies in many residency programs, earlier exposure to use of ultrasound is being achieved through informal activities embedded within medical school curricula. Using ultrasound to learn anatomy is now standard practice for many courses in the United States. However, very few if any offer formal opportunities to assess student skill and anatomical knowledge at the same time. As part of the Mini Ultrasound Skills Institute (MUSI) in Anatomy, a component of the first year anatomy course, students were expected to complete a team‐based, hands‐on ultrasound skills assessment. Throughout the course, students received both didactic briefing sessions and hands‐on training on ultrasound basics, image interpretation, and scanning techniques. The ultrasound content was presented in parallel to anatomy and radiology curriculum. In the past, ultrasound knowledge was assessed via static images; however, due to the contextual nature of ultrasound interpretation, a live, hands‐on method was devised to assess student learning. At regular intervals during the course, students completed an ultrasound skills assessment. In their dissection teams of four, students completed four tasks: setup the machine, obtained, oriented, and labeled a desired image using a live model. Each student was given a unique role within this exercise. Students were graded on a rubric scale for each of the four components and given an overall team score. At the conclusion of the course, students completed a graded ultrasound objective structural practical examination (OSPE). Rotating through four stations, students completed the four tasks on four different structures. Students rotated through roles within their team as they moved between stations. OSPE grade was incorporated into their final laboratory practical grade. Preliminary data from formative OSPE assessments ‐‐‐‐showed an average score of 72% (±21%) with a minimum of 50%, maximum of 100%. Within the skill specific breakdown, students were very comfortable at labeling obtained images (100%) and had the most challenging time with either selecting the correct probe or setting up the scanner 40% (±51%). Overall, team‐based OSPE is an effective method to promote and evaluate ultrasound skills learned in an anatomy course.
Microscopic Anatomy and Pathology Laboratories: Design of an E‐Learning Atlas as an Evolving Response to Interdisciplinary Medical Curricular Needs
Authors: Jurjus Rosalyn A. , Wade Jenna , Moktan Varun , Davis Michelle S. , Mills Alexandra , Butera Gisela , Krum Janette , Latham Patricia
Published: November 2023

Several institutions have integrated histology with other disciplines in team based laboratory approaches, but integration of histology and pathology instruction tools has been done to a lesser extent. Our aim was to develop an e‐learning atlas that integrates microanatomy and pathology laboratories for an interdisciplinary integrated medical curriculum. A multidisciplinary team is developing the online Microanatomy and Pathology Atlas (MAPA) (microanatomyatlas.com) that includes a library of histology and pathology images. Traditional laboratory manual instructions and study objectives are linked to specific labeled images that were made interactive to allow for self‐testing. Clinical case studies involving disease entities are being incorporated. Data is being collected on the number and frequency of medical students in the preclinical years using the atlas and a modified Likert‐style survey (1‐strongly agree to 4‐strongly disagree) is being used to assess student satisfaction and perceived learning. Preliminary data shows that 92%of students (score 1–2) would like to use the tool in the future, 62% strongly in favor (score 1). 92% of students (score 1–2) also stated they would advise their peers to use the MAPA. Data shows that the MAPA is positively correlated with material from lectures and is well suited for the level of student proficiency. In conclusion, MAPA is becoming an essential learning tool for students in our integrated curriculum at GWU‐SMHS.

Support or Funding Information

Supported by AEIUCM
Cyclophilin A is Crucial for Listeria and Salmonella Invasion and Cell‐to‐Cell Spreading
Authors: Dhanda Arandeep , Warren Kiera , Guttman Julian
Published: November 2023

The actin cytoskeleton is a common target for a variety of pathogens. Listeria monocytogenes (Listeria) bacteria control the actin polymerization machinery within eukaryotic cells for their entry, intracellular motility and subsequent spreading from one cell to another. These microbes use actin‐rich structures called comet tails to generate protrusions at the plasma membrane (referred to as listeriopods) for cell‐to‐cell spreading. Unfortunately, the precise molecular mechanisms governing this process remain elusive. Another actin‐hijacking bacterial pathogen is Salmonella typhimurium (Salmonella). These bacteria exploit actin‐based membrane ruffling in order to enter cells. Although actin regulators have been linked to this process, a thorough understanding of the molecular constituents and their functions at these structures has not yet been investigated. We have previously shown that the peptidyl prolyl cis‐trans isomerase (PPIase), cyclophilin A (CypA), a cytosolic protein involved in protein folding, is recruited to actin‐rich structures generated by Listeria. Here we expanded on our previous discovery and tested the hypothesis that CypA is important for the pathogenesis of Listeria. Infections of murine fibroblasts depleted of CypA revealed that listeriopods were structurally collapsed, suggesting a unique role of cellular PPIases during cell‐to‐cell spreading of Listeria. Importantly, when CypA was reexpressed in these cells, listeriopods appeared normal. Curiously, cytoplasmic comet tails remained morphologically unchanged when CypA was absent. We also found that CypA is recruited to actin‐rich membrane ruffles generated by Salmonella during its entry into cells. Surprisingly, invasion assays of murine fibroblasts showed that invasion by Listeria and Salmonella increases significantly in cells knocked‐out for CypA. These results suggest a novel membrane‐associated mechanism whereby CypA regulates the efficiency of invasion and cell‐to‐cell transmission of bacteria.

Support or Funding Information

Grant Funding Source: NSERC and departmental funds
Fos‐Immunoreactive Neurons In Rat Intrinsic Cardiac Ganglia After Pericardial Capsaicin Injection
Authors: Wang Ting , Miller Kenneth E.
Published: November 2023

Intrinsic cardiac ganglia (ICG) are composed of autonomic efferent neurons, local circuit neurons, interneurons, and sensory neurons. Some sensory neurons in the ICG express transient receptor potential vanilloid type1 (TRPV1). In the current study, capsaicin, a potent TRPV1 channel agonist, was applied pericardially to stimulate the afferent nerve terminals of sensory neurons of the rat ICG. Activated, post‐synaptic ICG neurons were evaluated by Fos‐immunoreactivity. Activated, glutamatergic ICG neurons were evaluated by immunostaining for Fos and for vesicular glutamate transporter 1 (VGLUT1) or glutaminase (GLS). The results of current study demonstrated that: 1) Pericardial capsaicin is able to induce Fos‐immunoreactivity in neurons of the ICG; 2) GLS and VGLUT1‐immunoreactivity is present within some of the activated ICG neurons. 3) Fos‐immunoreactive ICG neurons, with or without GLS and VGLUT1‐ immunoreactivity, occurred in two cell size populations, small (<300μm2) and large (>300μm2). These results provide further evidence that ICG are composed of local neuronal circuitry that participate in cardiac regulation.
Phantageusia: The Presence of Taste Distortions Inhibited by Repetitive Transcranial Magnetic Stimulation (rTMS)
Authors: Henkin Robert I , Abdelmeguid Mona , Knoppel Alexandra B , Potolicchio Samuel J
Published: November 2023

Phantageusia is an unpleasant, usually persistent obnoxious taste in the oral cavity which occurs in the absence of any oral stimulus and which affects many patients in the United States. It has been considered a type of sensory hallucination. It usually occurs after a pathological event such as an influenza‐like illness, head injury or Parkinson disease which causes loss of taste acuity. Magnetic resonance spectroscopy (MRS) of brain in patients with this symptom revealed a lower than normal level of brain gamma‐aminobutyric acid (GABA). We have studied 30 patients with phantageusia and treated them with rTMS with 20 stimuli at intervals of 1–5 sec applied repeatedly first with sham stimulation to shoulder, neck and head at an intensity of 0.2 T and then with full stimulation to the head only at an intensity of 1.1 T. Quantitative changes in phantageusia presence was determined after both sham and active rTMS on a 0–100 scale with 100 indicating complete phantageusia inhibition and 0 indicating no change in distortion. Sham stimulation elicited no change in phantageusia or its intensity. rTMS inhibited phantageusia in 24 of the 30 patients with a mean intensity decrease of 49%, complete inhibition of phantageusia in four patients and 90% inhibition in two others. Repeat MRS in patients treated successfully demonstrated significant increases in brain GABA over the untreated state. Phantageusia inhibition commonly lasted months‐years without recurrence. In some patients repeat rTMS was necessary to inhibit phantageusia more effectively. rTMS is an effective method to inhibit phantageusia related to increases in brain GABA.

Support or Funding Information

None
Let There Be Force: Modeling Cell‐ECM Interactions During Cancer Invasion
Authors: Jiang Yi
Published: November 2023

Extracellular matrix (ECM), the fibrous material that forms a network embedding cells in a tissue, significantly affects many aspects of cellular behavior, including cell movement and proliferation. Transgenic mouse tumor studies indicate that excess collagen, a major component of ECM, enhances tumor formation and invasiveness. Moreover, cell interactions with the collagen matrix result in aligned fibers that facilitate cell invasion. However, the underlying mechanisms are unclear, partially because the properties of ECM are complex, with diverse topographies and mechanical properties depending on various biophysical parameters. We have developed a three‐dimensional elastic computational fiber network model, and parameterized it with in vitro collagen experiments. The model can fill the gap between microscopic single collagen fiber studies and macroscopic collagen gel studies. We further developed a multiscale biomechanical cell migration model that interacts with the collagen network, and studied plasticity of cell migration in 3D ECM. This model is the first step toward a fully biomechanical cell‐matrix interaction model for changes in matrix organization during cell migration and tumor invasion.

Support or Funding Information

This work has been supported by the National Institute of Health 5U01CA143069.
A New Look at the Alveolar Macrophage
Authors: Bhattacharya Jahar
Published: November 2023

Severe lung inflammation can lead to life‐threatening conditions such as acute lung injury (ALI) and the Acute Respiratory Distress Syndrome (ARDS). Alveolar macrophages are critical to these mechanisms. Not only do they clear debris, but as immune sentinels they direct the lung's pro‐ and anti‐inflammatory responses to optimize gas exchange. Despite the importance of macrophages in lung biology, a major difficulty is that the bulk of our understanding of lung macrophage function comes from studies on cells isolated from lung tissue or the bronchoalveolar lavage (BAL). Since cell isolation may change cell function, studies of macrophages in intact alveoli are required.

Live‐lung imaging studies provide a means for direct studies of alveolar macrophages in situ. The studies reveal that resident alveolar macrophages adhere to the alveolar epithelium (AE) and resist washout. These cells are called sessile alveolar macrophages (SAMs) to distinguish them from macrophages that can be washed out and recovered in the bronchoalveolar lavage (BAL), as well as from lung macrophages that are derived from adult bone marrow‐derived monocytes. About half of the SAMs form stable gap‐junctional channels (GJCs) with the AE. Remarkably, SAMs signal through these GJCs to suppress the lung's exuberant inflammatory response to lethal endotoxin challenge.

Pathogen detection mechanisms by SAMs remain unclear. One possibility is that pathogen detection involves the alveolar wall liquid (AWL). The alveolar distribution of SAMs is approximately 1 per 3 alveoli. Moreover, SAMs are non‐migratory. Hence it is possible that pathogen detection occurs when AWL flow convectively delivers the pathogens to SAMs. Chloride secretion by the alveolar cystic fibrosis transmembrane conductance regulator (CFTR) causes AWL flow. The flow is outward from alveoli towards the alveolar duct. In mice bred to specifically delete CFTR in alveolar type 1 (AT1) epithelial cells, AWL secretion stops and alveolar clearance of bacteria is decreased. These data support the hypothesis that pathogen delivery through AWL flow generated by AT1 cells induces proinflammatory responses in SAMs. Thus, alveolar CFTR may determine SAM‐induced pro‐ or anti‐inflammatory mechanisms in ALI.

Support or Funding Information

Supported by NIH grant HL57556.