bluuebunny/crossref_metadata_2025 huggingface dataset:
Spatial organization of signaling in bacterial chemotaxis
Authors: Sourjik Victor
Published: June 2021

Spatial organization of signaling is not an exclusive property of eukaryotic cells. Despite the fact that bacterial signaling pathways are generally simpler than those in eukaryotes, there are several well‐documented examples of higher‐order intracellular signaling structures in bacteria. One of the most prominent and best characterized structures is formed by proteins that control bacterial chemotaxis. Signals in chemotaxis are processed by ordered arrays, or clusters, of receptors and associated proteins, which amplify and integrate chemotactic stimuli in a highly cooperative fashion. Receptor clusters further serve to scaffold protein interactions, enhancing the efficiency and specificity of the pathway reactions and preventing formation of signaling gradients through the cell body. Assembly of receptor clusters appears to be a stochastic process, but bacteria evolved mechanisms to ensure optimal cluster distribution along the cell body for partitioning to daughter cells at division. Finally, recent findings demonstrate that receptor clusters can adapt their structure, function and dynamics of protein exchange to ensure optimal signal processing under particular environmental conditions.
Metabolic engineering of Saccharomyces cerevisiae for alkaloid production
Authors: Smolke Christina Dawn
Published: June 2021

Natural products or natural‐product inspired molecules comprise most successful drugs. However, these high‐value molecules are difficult to synthesize chemically and are often isolated through inefficient extractions from natural sources. As such, they are increasingly targeted for production by biosynthesis from engineered microorganisms. The budding yeast species Saccharomyces cerevisiae has proven to be a powerful microorganism for heterologous expression of biosynthetic pathways. Yet many challenges are currently faced in engineering yeast strains for the biosynthesis of complex secondary metabolite production. Synthetic biology is advancing the development of new tools for constructing, controlling, and optimizing complex metabolic pathways in yeast.

I will describe recent work in engineering yeast as a production host for diverse alkaloid compounds, and in particular the benzylisoquinoline alkaloids (BIAs). Yeast strains engineered to express enzymes from diverse organisms, including plants, bacteria, and humans, have been demonstrated to synthesize numerous intermediates and end‐products in this biosynthetic pathway. In addition, I will describe the development of synthetic biology platforms that are providing new approaches to optimizing complex biosynthetic pathways.
Pyrroloquinoline Quinone (PQQ) and Indicators of Antioxidant Potential, Inflammation and Metabolism in Humans
Authors: Rucker Robert B , Harris Calliandra , Chowanadisai Winyoo , Slupsky Carolyn
Published: June 2021

Pyrroloquinoline quinone (PQQ) improves parameters related to energy metabolism in animals, which involve interactions with cell signaling important to mitochondrial function. However, little is known about responses to PQQ in humans. Using a crossover study design, 10 subjects ingested PQQ and were examined in two studies at 2 to 3 daily intervals. Plasma antioxidant potential (TBARS and TRAP measurements), inflammation (plasma C‐reactive protein and IL‐6), and standard clinical indices were measured, including urinary metabolites. In addition, PQQ clearance was investigated. Data suggested that PQQ is absorbed in the upper intestine with a relatively slow clearance from the plasma pool. Measurement of plasma antioxidant activity was highly correlated with plasma PQQ concentrations. Decreased levels of plasma C‐reactive protein and IL‐6 were also observed (~ 20 mg PQQ/day X 3 d; p <0.01). No changes in clinical indices were observed. However, exposure to PQQ (10–20 mg PQQ/day X 3 d) altered the distribution of mitochondrial‐related urinary metabolites in urine using as assessed by the NMR. It was concluded that the effects of PQQ in humans could be observed with a short‐term exposure at levels easily obtained in commercial supplements. Supported by Mitsubishi Gas and Chemical Co. and the Center for Health Related Research at UCD.
Purified diets are obesigenic, promote insulin resistance and confer sensitivity to chemical colitogens
Authors: Aitken Jesse D , Vijay-Kumar Matam , Srinivasan Gayathri , Gewirtz Andrew
Published: June 2021

The paradigm that an obesigenic metabolic profile results from excess caloric intake has come under question as a growing number of caveats, especially that predisposition to insulin resistance and obesity is heritable, through epigenetic and microbiotal mechanisms, and self‐perpetuating, as adipose tissue is both a source and sink for inflammatory actors, emerge. As obesity penetrance increases in developing countries on adoption of Western diet and food processing regimes, it seems logical to investigate the obesigenic potential of highly processed, although nominally chow‐equivalent, diets in mice. Naturally sourced chow and purified 10% saturated fat diets were given to 4 week old mice whose weights were followed for 12 weeks at which time metabolic and inflammatory parameters were assessed. Further, chow‐ and purified diet‐fed mice were subjected to chronic and acute models of DSS colitis. We report that purified diets induce weight gain and insulin resistance relative to isocaloric naturally‐sourced chow. Purified diet also induced mild splenomegaly and wasting of intestinal tissues accompanied by increases in neutrophil infiltration, proinflammatory cytokines and acute phase proteins, perhaps reflecting diet‐mediated degradation of barrier function. Purified diet‐fed mice were sensitive to DSS‐induced mortality and weight loss immediately following co‐administration of purified diets.

Grant Funding Source : NIH
In vivo mouse model for examining contribution of inflammation to development of obesity and diabetes
Authors: Komarnytsky Slavko , Esposito Debora , Raskin Ilya
Published: June 2021

An adipose tissue‐mediated shift to production of proinflammatory cytokines in obesity likely contributes to the low‐level systemic inflammation that is seen in metabolic syndrome‐associated chronic pathologies such as diabetes. The mechanism explaining this relationship remains unknown. In this study, we present specific details on using pharmacological supplementation with triptolide, a diterpene triepoxide with anti‐inflammatory and immunosuppressive properties, to diet‐induced obese (DIO) C57BL/6J male mice. Animals received 50 ug/kg of triptolide in corn oil (vehicle) via oral gavage every 3 d for 9 wk and underwent oral glucose tolerance test on wk 1 (baseline), wk 7 and wk 9. Significant increase in glucose utilization and improved insulin sensitivity were associated with triptolide in the absence of the direct effects on obesity measures of food intake, body weight, or composition. Macrophage recruitment and cytokine signature of the adipose tissue were significantly reduced by triptolide with minimal or no effect on the adipose humoral factors leptin and adiponectin. These data demonstrate that DIO‐triptolide mouse is a tractable animal model for the study of obesity‐related metabolic dysfunction and chronic inflammation. The study was supported in part by the 5P50AT002776‐05 grant from NCCAM.
Adipocytes from obese subjects increase NF‐kB activation in invasive breast cancer cells
Authors: Siriwardhana Nalin , Karwandyar Ayub , Wimalasena Jay , Moustaid-Moussa Naima
Published: June 2021

Obesity in post‐menopausal women doubles the breast cancer risk and metastasis. Concomitantly, nuclear factor kappa B (NF‐κB) is known to play a significant role in breast cancer cell metastasis. We used a novel three dimensional (3D) co‐culture system with breast cells (non‐cancerous, MCF10A, cancerous, MCF7 and invasive, MDA231) and adipocytes from pre‐ and post‐menopausal women in lean and obese groups. Comparison of cytokine profiles of co‐cultures revealed two‐way interaction of adipocytes and breast cells. Highly significant cytokine changes were reported for post‐menopausal adipocytes and MDA MB 231 co‐culture. Therefore, we assayed the NF‐κB activation effects of lean and obese post‐menopausal adipocytes on MDA MB 231 cells. Media transfer from lean/obese post‐menopausal adipocyte cultures in to growing MDA MB 231 breast cells resulted significant changes in NF‐κB activation pathway genes. This further verified our previous data that obese conditioned media increased the levels of NF‐κB family members (P65, P50, P52, Rel B and Rel C) compared to the lean counterpart. Therefore, our data indicates that adipocytes from post‐menopausal obese women can increase the NF‐κB activation in MDA MB 231 cells. In conclusion, this study suggests that increased breast cancer risk in obese post‐menopausal women may be at least in part due to obesity mediated NF‐κB activation. Support: UTORC, AgResearch and UTGSM.

Grant Funding Source : Internal
Comparative Proteomic Profiling of Human Breast Cell Lines
Authors: Turbyville Tommy J , Blonder Josip , Das Sudipto , Ye Xiaoying , Prieto DaRue A , Veenstra Timothy D , Milner John A , Romagnolo Donato F
Published: June 2021

Although epidemiological studies have suggested that the isoflavone genistein found in soy may be protective against breast cancer, its effects on molecular processes involved in cell proliferation have not been clearly elucidated. Using comparative shotgun proteomics, we examined the proteome of non‐cancerous breast MCF10A cells, estrogen receptor‐positive breast cancer T47D cells, and Hras transfected MCF10A cells after treatment with 3, 10, and 30 μM genistein for 18 h. While genistein induced G1‐phase arrest in MCF10A‐Hras and G2/M‐phase arrest in T47D cells, it did not alter the cell cycle distribution in MCF10A cells. The arrest in G1‐phase of MCF10A‐Hras cells coincided with downregulation of the proliferation marker Ki67, epidermal growth factor receptor, various components of the mitogen‐activated protein kinase family of enzymes and low molecular weight phosphotyrosine protein phosphatase (ACP1), The arrest in G2/M‐phase of T47D cells was accompanied by upregulation of caspase‐8 and mSH2, suggesting that genistein may induce a G2/M checkpoint/proapoptotic response involving the mismatch repair system. Taken together, these findings suggest a differential response to genistein in cancerous cells and non‐malignant epithelial cells, and demonstrate the utility of comparative shotgun proteomics for investigating the cancer preventative effects of bioactive components.
Effect of (−)‐epigallocatechin gallate on cyclin D1 down‐regulation at the post‐translational level
Authors: Zhang Xiaobo , Baek Seung Joon
Published: June 2021

The potential anti‐carcinogenic effect of green tea catechins has been well documented, especially for one of the major components, (−)‐epigallocatechin gallate (EGCG). Cyclin D1, an oncogenic protein, has been shown to be up‐regulated in various cancer cells, including colorectal cancer. It has been reported that EGCG inhibited cyclin D1 at the transcriptional and translational levels; however, the detailed mechanisms by which EGCG suppresses cyclin D1 expression have not been elucidated. Our objective was to elucidate the molecular mechanism(s) and biological consequence of EGCG‐induced cyclin D1 down‐regulation in colorectal cancer cells. We examined the effect of EGCG on the expression of cell growth related proteins in colorectal cancer cells. As a result, we found that EGCG inhibited cell growth and induced cell cycle arrest and apoptosis. The expression of cylin D1 and D3, but not cyclin E was down‐regulated, while p21 was up‐regulated in the presence of EGCG. Moreover, the cyclin D1 down‐regulation by EGCG was mediated by the 26S proteasome pathway dependent of glycogen synthase kinase‐3 beta (GSK‐3β). Collectively, EGCG facilitates the proteasomal degradation of cyclin D1, which could, at least in part, be responsible for the anti‐cancer effect of EGCG in colorectal cancer.

Supported by: NIH R01CA108975 and The University of Tennessee Center of Excellence in Livestock Diseases and Human Health grant

Grant Funding Source: NIH R01CA108975
Release 2 of the U.S. Dietary Supplement Ingredient Database (DSID): research protocols and ingredient estimates for children's and adult multivitamins
Authors: Andrews K W , Roseland J M , Holden J M , Middleton A M , Solomon A M , Douglass L , Dwyer J T , Bailey R , Saldanha L G , Daniel M G
Published: June 2021

The Dietary Supplement Ingredient Database (DSID) is a federal initiative to provide analytically‐derived estimates of ingredients in dietary supplements because the actual content may differ from the labeled amount. Release 2 provides data for children's multivitamins (MVMs), updates adult MVM estimates and is available at http://dsid.usda.nih.gov. Representative children's MVMs were identified and purchased from multiple market channels. These supplements were analyzed for their vitamin and mineral content with certified reference materials. Mean percent differences from label were calculated for each nutrient and compared to labeled levels using regression analysis. The relationship between label and analytical content for each nutrient was identified as either linear (n=12) or quadratic (n=5). Mean % differences from label averaged 1 to <10% above label for eight nutrients (zinc, phosphorus, iron, magnesium, copper, vitamin B‐6, niacin, riboflavin); 10 to <20% above label for 5 nutrients (thiamin, folic acid, vitamin B‐12, manganese, and calcium); 20 to <30% above label for 3 nutrients (iodine and vitamins A and E) and >30% above label for vitamin D. Data estimates were linked to children's MVM products reported in the National Health and Examination Survey and can be used by researchers to more accurately quantify nutrient intake from dietary supplements. Funding: ARS/USDA & ODS/NIH Y1CN501006
Change in eating behaviors in a 6 month pilot worksite weight loss intervention
Authors: Batra Payal J , Salinardi Taylor , Robinson Lisa , Saltzman Edward , Roberts Susan , Das Sai K
Published: June 2021

The eating behavior constructs restraint, hunger and disinhibition may play an important role in successful weight control. However, the extent to which these constructs can be influenced to sustain weight loss during an intervention remains uncertain. The objective of this pilot study was to determine whether a weight loss intervention at worksites would cause significant changes in eating behaviors. A 6‐mo randomized controlled trial was conducted at 4 worksites [2 intervention (I), 2 control(C)] in the Boston metropolitan area. Measures included fasting body weight and eating behavior assessed using the Eating Inventory at baseline and 6‐months. Results were analyzed using an ANCOVA model with baseline scores as covariates. There were significant increases in restraint (I=5.43Â±4.25, C=0.29Â±3.80,p<0.001), decreases in disinhibition (I=−2.5Â±3.63, C=0.66Â±1.85, p<0.001) and hunger scores (I=−2.79Â±3.13, C=0.56Â±2.63, p<0.001) in the I compared to the C group. Within the I group, decrease in hunger is the strongest predictor of weight loss, the model contributing to 32% of the variance (p<0.001) after controlling for gender and site. This is in contrast to previous studies that have shown change in restraint to be the strongest predictor of weight loss. These findings identify reduction in hunger as an important predictor of successful weight loss.

Grant Funding Source: HNRCA
High‐dose (4000 IU) vitamin D supplementation improves insulin resistance in obese adolescents
Authors: Belenchia Anthony M. , Tosh Aneesh K , Hillman Laura S , Peterson Catherine A.
Published: June 2021

Obese adolescents with vitamin D (vit D) deficiency have been observed to have greater body weights and exhibit early signs of insulin sensitivity and higher risk of developing type 2 diabetes. Our objective was to determine the effect of high‐dose vit D supplementation on insulin resistance in this population. Forty‐four male and female patients (age=13.9±2.5 years; BMI Z‐score= 2.52±0.25) were recruited from the University of Missouri‐ Adolescent Diabetes and Obesity clinic and were randomized to receive either placebo or vit D3 (4000 IU/day) as part of their standard treatment for 6 months. At the start, all but three patients were vit D deficient (<20ng/ml) or insufficient (20–29 ng/ml). Within 3 months of supplementation, none of the subjects taking vit D were “deficient”, and by 6 months, 93% had 25‐ hydroxyvitaminD levels considered “sufficient”. Likewise, adolescents receiving vit D had significant improvements in both homeostasis model assessment‐estimated insulin resistance (HOMA‐IR, −1.86 vs +0.22 for placebo; P=0.037); and quantitative Insulin Sensitivity Check Index (QUICKI, +0.017 vs −0.003; P=.036). Correcting the poor vit D status associated with obesity may be an effective and inexpensive adjuvant to treatment of obesity‐related metabolic complications.

Grant Funding Source : Support: J.R. Albert Foundation, Inc.
Vitamin D Status in Relation to Metabolic Risk Factors in Older Puerto Ricans
Authors: Jamal-Allial Aziza , Homsi Yamen , Tucker Katherine L
Published: June 2021

Vitamin D deficiency is a global problem. Several reports suggest an association between plasma 25‐hydroxy vitamin D concentration (25(OH)D) and metabolic risk factors. We examined, corss‐sectionally, the relation between serum 25(OH)D concentration and adiposity and glucose metabolism among 1505 Puerto Rican adults, 484 men and 1058 women aged 45–75 years. Among all participants, body mass index (BMI), glycosylated hemoglobin (HbA1c), insulin resistance (HOMA‐IR), waist circumference (WC) and waist‐to‐hip‐ratio (WHR) showed significant inverse association with plasma 25(OH)D (P<0.05); but diabetes status and insulin concentration did not. After stratification by sex, 25(OH)D was significantly associated with BMI, HbA1c, HOMA‐IR, WC, and insulin concentration (P<0.05) in women. Among men, only HbA1c and WHR showed constant inverse association with plasma 25(OH)D (P<0.05). Women with BMI ≥ 35 or BMI 30–35, respectively, had lower serum 25(OH)D concentrations than women with BMI <25 (P<0.01 and P<0.001 respectively and p‐trend=0.001). In conclusion, lower serum 25(OH)D concentration was significantly associated with adiposity measurements and metabolic risk factors in this population of older Puerto Rican adults living in the Boston area. Longitudinal studies are needed to clarify causal pathways.

This study was supported by the National Institutes of Health grants P01 AG023394 and P50 HL105185.

Grant Funding Source: NIH
Iron biomarkers identify similar risk factors for iron deficiency but provide different estimates of the national prevalence of iron deficiency and iron‐deficiency anemia in Cameroon
Authors: Engle-Stone Reina , Nankap Martin , Ndjebayi Alex O. , Erhardt Juergen G. , Brown Kenneth H.
Published: June 2021

Iron status indicators reflect different aspects of iron metabolism. We compared the prevalence and distribution of iron deficiency (ID) and iron‐deficiency anemia (IDA) in a national survey of Cameroonian women and children using plasma ferritin (F), soluble transferrin receptor (sTfR), body iron stores (BIS), and hemoglobin (Hb), with and without adjustments for inflammation. We randomly selected 30 clusters in each of 3 zones (North, South, and Large Cities) and 10 households (HH) per cluster, each with a woman 15–49 y and a child 12–59 mo (n=904 HH). Plasma F and BIS were adjusted for inflammation using C‐reactive protein and α1‐acid glycoprotein. Inflammation was present in 21% of women and 48% of children; 39% of women and 58% of children were anemic. The prevalence of ID ranged from 12–32% among women and 14–68% among children, depending on the iron status indicator, and IDA ranged from 9–19% among women and 12–47% among children. The proportion of anemia associated with ID ranged from 26–50% among women and 21–82% among children. However, the indicators identified similar groups at highest risk of ID and IDA: HH in the North and rural areas and younger children and pregnant women. SES was less consistently related to ID. Research is needed to clarify relationships among iron indicators, particularly during infection, to harmonize global data on ID and IDA.

Grant Funding Source : Michael and Susan Dell Foundation, UNICEF
Paraoxonase 1 (PON1) as a marker of short term death in breast cancer recurrence
Authors: Bard Jean-Marie , Jaffré Isabelle , Joalland Marie-Pierre , Classe Jean-Marc , Campone Mario , Bobin-Dubigeon Christine
Published: June 2021

Objective

To relate paraoxonase (PON1) activity to survival time and short term death in breast cancer recurrence.

Design and methods

PON1 activity was measured by its rate of hydrolysis of two different substrates, paraoxon (PON) and phenylacetate (ARE) in 50 patients who experienced disease recurrence for breast cancer. Results were compared between patients surviving more than one year after the analysis (n=22) and those who died within one year (n=28).

Results

In a logistic regression analysis, ARE was significantly and negatively associated with early death (OR = 0.10 [0.02–0.58], p=0.0109). PON did not reach significance (OR = 0.43 [0.17–1.11], p=0.0826). As assessed by the Kaplan‐Meier method, the survival distributions across quartiles of ARE was significantly different (Log rang test, p=0.0393). The Cox proportional hazards model indicated that quartile 1 of ARE was significantly different from quartile 4 (HR= 5.48 [1.48–20.36], p=0.0110. In a multiple logistic regression analysis model, ARE was independently associated with early death (OR = 0.12 [0.02–0.98], p=0.0476), besides interval time between diagnosis and recurrence (OR = 0.54 [0.27–1.07], p=0.0781) and undernutrition (OR = 3.95 [0.81–19.19], p=0.0883).

Conclusion

Paraoxonase activity may predict survival in patients with breast cancer recurrence.
Formative Research Supports a Change Agent Approach to Prevent Obesity in American Indian Households
Authors: Gadhoke Preety , Christiansen Karina , Rowan Megan , Anliker Jean , Frick Kevin , Gittelsohn Joel
Published: June 2021

High obesity rates among Americans Indians are associated with environmental factors that influence food and physical activity‐related behaviors. Formative research in Michigan consisted of 98 interviews (workshops, in‐depth, focus groups, and household groups). The AI household is a key influential environment upon health behaviors. Children as change agents can impact household diets and physical activity by “sharing” their learned knowledge from school, and by “encouraging,” “reminding,” and “asking” for healthier foods and to be physically active with them. Adults “listen” to and are “inspired” by children's “energy.” Intervention trials for AI communities will use a novel child as change agent approach for improving health behaviors among adult household members. USDA/NIFA/NRI, Grant #2010‐85215‐20666.

Grant Funding Source : USDA/NIFA/NRI, Grant #2010‐85215‐20666
Is fruit and vegetable intake of preschool children associated with parental feeding practices?
Authors: Shim Jae Eun , Kim Juhee ,   The STRONG Kids Research Team
Published: June 2021

This study was performed to examine the association between parental feeding practices and preschooler's fruit and vegetable intake. Cross‐sectional analyses of baseline data from 230 mothers who enrolled in the cohort of STRONG Kids program were conducted. Ten aspects of parental feeding practices were measured using a Comprehensive Feeding Practices Questionnaire and frequency of child's fruit and vegetable consumption was estimated by mother. The children in the study consumed fruits 1.42±0.77 times a day and vegetables 1.73±0.90 times a day. The results showed that children's fruit consumption was evenly associated with “encourage balance and variety,” “modeling,” “monitoring,” and “environment.” Vegetable consumption was most correlated with “environment” followed by “encourage balance and variety” and “modeling” indicating that associated feeding practices are different from those of fruit consumption. “Emotion regulation,” “restriction for health,” “food as reward” and “pressure” had no associations with child's fruit and vegetable consumption. The results suggested the importance of warm emotional interaction during feeding events and building an environment favorable to fruit and vegetable consumption in preschool aged children. Longitudinal studies are warranted to understand the causal relation between parental feeding practices and child's eating behaviors.
Lactobacillus colonization induces ROS‐dependent intestinal development
Authors: Jones Rheinallt M. , Luo Liping , Lin Patricia W. , Neish Andrew S.
Published: June 2021

Commensal bacteria exhibit positive influences on intestinal homeostasis. However, the molecular mechanism(s) that mediate these beneficial responses remain uncharacterized. Recently, multiple lines of investigation have shown the rapid cellular generation of physiological levels of reactive oxygen species (ROS) in response to Lactobacillus contact with cultured epithelial cells. Here, we recapitulate these observations in vivo using newly emerging germ‐free Drosophila first instar larvae, newly colonized 2‐day‐old mice, or adult germ‐free mice. L. plantarum or L. rhamnosus GG colonization of germ‐free flies or mice induced the rapid generation of cellular ROS, and ROS‐dependent cellular proliferation. Mechanistically, we genetically modulated ROS levels within specific cells in the intestinal stem cell (ISC) microenvironment of Drosophila. ROS generation within ISC progenitors markedly enhanced proliferation and differentiation, whereas suppressing ROS inhibited development. Furthermore, suppressing ROS significantly increased the numbers of undifferentiated cells within the Drosophila ISC niche. Together, these data show that the signaling pathways that control ISC proliferation and differentiation are sensitive to Lactobacillus‐induced ROS generation, and highlight a molecular mechanism whereby commensal bacteria influences intestinal development.
Antisense oligonucleotide therapy: combating aberrant β‐catenin in hepatocellular carcinoma using peptide nucleic acids without transfecting agents
Authors: Delgado Evan R , Bahal Raman , Ly Danith , Monga Satdarshan Pal
Published: June 2021

Patients in the United States with Hepatocellular carcinoma (HCC) have roughly a 13% chance of survival. HCC is the 3rd leading cause of death by cancer worldwide and lack of efficacious treatment remains a major concern. Wnt/β‐catenin signaling has been shown to be increased in around 30% of all cases of HCC due to stabilizing mutations in the β‐catenin gene. Here we report development of a novel, cell permeable, guanidine‐based, peptide nucleic acid (PNA) antisense oligonucleotide system designed against either the transcription start site (T1) or the translation start site (T2) of the human β‐catenin gene. Cell uptake efficiency was monitored by a fluorescent TAMRA tag on the PNA by live cell imaging. Both T1 and T2 passed comparably into HCC cells without aid of any transfection reagents. T1 and T2 inhibited β‐catenin activity in HepG2 cells that harbor mutant β‐catenin as measured by TOPflash luciferase reporter, but T1 dominated over T2. T1 at nano‐molar concentration decreased β‐catenin activity in HepG2 cells. Real Time‐PCR confirmed a 55% reduction in β‐catenin mRNA. Lysates from T1 treated HepG2 cells showed a significant decrease in total β‐catenin protein as well as downstream targets associated with β‐catenin activity such as Cyclin‐D1, Glutamine Synthetase and others. Decreases in HCC cell proliferation and viability was also evident following T1 treatment. Thus we report a novel modality that targets β‐catenin signaling in HCC.
Reduced ABC efflux transporter function in activated microglia: Implications in neurodegeneration
Authors: Gibson Christopher , Richardson Jason R. , Aleksunes Lauren M.
Published: June 2021

Efflux transporters in the brain limit exposure to chemicals and potentially toxic endogenous products, which may be altered during neurodegeneration and contribute to further injury. The purpose of this study was to characterize ABC efflux transporter function in unstimulated mouse BV2 microglia and in microglia activated with lipopolysaccharide (LPS, E.coli). Fluorescent substrates and known inhibitors were used to characterize the function of efflux transporters Resting microglia actively transported rhodamine 123, calcein AM, and Hoechst, which were blocked by inhibitors of MDR1 (PSC833), MRP (MK571), and BCRP (KO143), respectively. Activation of microglia with LPS significantly reduced transporter function, indicated by up to a 90% increase in cellular retention of rhodamine 123, Calcein AM, and Hoescht. Decreased efflux transporter function in activated microglia correlates with down‐regulation of transporter mRNA and protein levels. Collectively, these results suggest that microglia activation during neuroinflammation may impair normal regulatory functions of microglia including the efflux of potentially toxic compounds. NIH ES‐0205022, ES‐005022, ES‐015991, DK‐ 080774, ES‐007148.
Thymoquinone inhibition of proinflammatory cytokines and nitric oxide production in LPS‐activated BV2 microglia cells
Authors: Taka Equar , Mazzio Elizabeth A , Goodman Carl B. , Soliman Karam F.A.
Published: June 2021

The immune responses elicited by proinflammatory cytokines and subsequent nitric oxide (NO) generation in the brain have been implicated in several neurodegenerative diseases such as Alzheimer disease and Parkinson's disease. To investigate the neuroprotectant action of thymoquinone (TQ) in activated microglia, we measured cytokines profile and nitric oxide (NO) production in LPS‐stimulated microglia cells in the presence or absence of TQ. TQ is the main active constituent of oil extracted from the Nigella sativa plant seeds. We hypothesized that the anti‐inflammatory properties of TQ are mediated in part by inhibiting the formation of pro‐inflammatory cytokines, chemokines and other inflammatory mediators. In this study, the microglia BV2 cells were stimulated with LPS in the presence or absence of TQ. To profile the effect of TQ on pro‐inflammatory molecules, RayBio cytokine antibody array was used. The results of this study showed that TQ totally inhibited IL‐6 production, resulted in 34% inhibition of IL‐12p40/70, inhibited growth colony stimulating factor (GCSF), and resulted in complete inhibition of the pro‐inflammatory chemokine MCP1 and the complete inhibition of NO production in LPS‐stimulated BV2 cells. These findings suggest that TQ may be useful in delaying the onset of progression of neurodegenerative disorders involving activated microglia cells. (Supported by a NIH grant RR 03020)
MMP‐9 overexpression in macrophages regulates the post‐myocardial infarction inflammatory response through SCYE1
Authors: Zamilpa Rogelio , Ramirez Trevi A , Dai Qiuxia , Dayah Tariq , Nguyen Nguyen , Zhang Jianhua , Ahuja Seema S , D'Armiento Jeanine , Jin Yu-Fang , Lindsey Merry L
Published: June 2021

Post‐myocardial infarction (MI), proinflammatory macrophages secrete matrix metalloproteinase (MMP)‐9 to regulate the cardiac wound healing response. We hypothesized that overexpression of MMP‐9 in macrophages would amplify the inflammatory response to exacerbate left ventricular dysfunction post‐MI. Accordingly, we assessed function of the left ventricle (LV) in wild‐type mice (WT, n=16) and MMP‐9 macrophage‐specific transgenic mice (TG, n=28) at day 5 post‐MI. In response to similar infarct sizes of 49±2% for both WT and TG (p=0.9), LV ejection fraction was improved in the TG (25±2%) compared to the WT mice (18±2%; p<0.05). We also evaluated macrophage numbers by Mac‐3 and galectin‐3 levels and inflammatory cytokine gene expression. Despite a similar number of macrophages in both WT and TG, the expression of 23 inflammatory genes decreased in the infarct from the TG LV compared to WT LV (all p<0.05). Interestingly, SCYE1, a multifunctional cytokine involved in the regulation of inflammatory responses, was uniquely upregulated at baseline in TG (0.92±0.13 2−ΔCT units) compared to WT (0.52±0.07 units; p<0.05). SCYE1 levels decreased post‐MI in the TG (0.52±0.08 units) while WT levels doubled (1.02±0.16 units; p<0.05). The decrease in the TG levels of SCYE1 suggest that SCYE1 is a key MMP‐9 responsive regulator of the inflammatory response that follows MI.
miR150‐knockout prevents Fas‐induced liver injury via Akt pathway
Authors: Chen Weina , Han Chang , Wu Tong
Published: June 2021

MicroRNA 150 has been reported to involve in a diversity of cellular process such as hematopoiesis, cell differentiation, proliferation, and apoptosis. However, its potential implication in liver injury has not been investigated. In this study, we use miR150 knockout (KO) and wild type (WT) mouse models to investigate the effect of miR150 in Fas‐induced hepatocyte apoptosis and liver injury in vivo. The wild type (WT) mice and miR150 knockout (KO) mice were treated with anti‐Fas antibody Jo2 (0.5μg/g of body weight) for 2 to 4 hours, and the extent of liver injury was assessed by histopathology, serum aminotransferases, TUNEL staining, and caspase activation. The miR150 KO mice showed resistant to Fas‐induced liver injury in comparison to WT mice; this was reflected by the lower alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, less liver damage, and less hepatocyte apoptosis (P<0.01). After Jo2 treatment, the protein and phosphorylation levels of AKT (the bona fide predict target of miR150) significantly decreased in WT mice but not in miR150 KO mice. Pretreatment with Akt inhibitor V (an Akt inhibitor) exacerbated Jo2‐mediated liver injury in miR150 KO mice. Furthermore, at lower Fas agonist dose (0.35μg/g of body weight), miR150 KO mice developed mild liver injury and all the treated mice survived, whereas WT mice showed severe liver injury and died within 10 hours. Taken together, these findings demonstrate that miR150 deficient prevents Fas‐induced hepatocyte apoptosis and liver injury and this protection is partly mediated by Akt‐induced signaling transduction pathway.
LINEAR GROWTH IN VERY PREMATURE INFANTS IS SIGNIFICANTLY RELATED TO PROTEIN INTAKE
Authors: Berseth Carol Lynn , Harris Cheryl , Mitmesser Susan Hazels
Published: June 2021

Background

We have previously shown that preterm infants fed human milk fortified with a liquid human milk fortifier (LHMF), which provides approximately 20% more protein than current powder fortifier (Control), have significantly greater linear growth. Previous studies have shown that preterm growth is a function of parenteral protein intake; however, this relationship for enteral protein has not been confirmed.

Objective

To assess linear growth as a function of protein intake.

Methods

Fifty‐six preterm infants (BW ≤1,250 g) fed expressed and/or donor breast milk were randomized to receive Control or LHMF for 28 days. Length and total protein intake (parenteral and enteral) were recorded.

Results

On study day 28, LHMF fed infants were significantly longer than Control (42.4±0.28 cm vs. 41.2±0.24 cm, p=0.003). There was no difference between groups for total calorie intake (116.0±1.5 kcal/kg/d vs. 119.3±1.3 kcal/kg/d, p=0.113). The change in length in all infants was significantly related to total protein intake (p=0.0077) (see Figure); a similar relationship was not present for nonprotein calorie intake (p=0.12). This relationship was also dependent upon the fortifier consumed (p=0.0327).

Conclusion

Linear growth in preterm infants reflects protein intake regardless of route of delivery. The better length growth seen in LHMF infants appears to be due to the higher enteral protein it provides.
Exposure to Simulated Microgravity Affects Development of the Skull in Danio rerio
Authors: Franz-Odendaal Tamara Anne , Edsall Sara
Published: June 2021

To determine how exposure to simulated microgravity (SMG) affects development of the skull, zebrafish embryos were exposed to SMG starting at one of three developmental stages: 10, 12 or 14 hours post‐fertilization. Juvenile and adult fish were analyzed after exposure to SMG using whole‐mount staining and geometric morphometrics. Fish were examined for changes in melanophore surface area and number, in addition to changes in skull morphology. Analyses reveal an overall decrease in the surface area of melanophores and an increase in melanophore number over the period of a week in experimental and control groups; with significant differences in the number of melanophores on the first day after exposure. These effects are short‐term and do not persist in adults. Gross morphological analyses of whole‐mount stained adults, however, identified buckling in several (but not all) craniofacial bones. In conclusion, exposure to SMG affects both skeletal tissues and pigmentation in the head. These results will be discussed in context of germ layer origin. This is the first study examining the long‐term effects of exposure to SMG on zebrafish and provides valuable insight into phenotypic variation.

Grant Funding Source : NSERC, Canada; and CSA, Canada
Create an engineered conduction tissue by seeding cardiac progenitor cells on gelatin foam
Authors: Zhang Chuansen , Zhang Xi , Yang Xiangqun
Published: June 2021

CompleteAV conduction block is typically treated by implanting an electronicpacemaker device; however, long‐term pacing therapy in pediatricpatients has significant complications. In this study, an engineered conduction tissue seeded with cardiac progenitor cells were implanted in rat hearts to create an alternative AV conduction pathway. The engineered conduction tissue was created by seeding cardiac progenitor cells on gelatin foam and exhibited sustained electrical coupling through persistent expression and function of gap junction proteins and pacemaker genes. Cardiac progenitor cells in the constructs were shown to survive in the hearts and the coupling of grafted cells with host cardiomyocytes was shown. Our experiments suggest the possibility that engineered tissue constructs can function as an electrical conduit and, ultimately, may offer a substitute treatment to conventional pacing therapy.
The Trapezius Complex in Mammalia
Authors: Voegele Georgina McKusick
Published: June 2021

Within Mammalia, the five portions of the trapezius complex ‐ sternomastoideus, cleidomastoideus, clavotrapezius, acromiotrapezius, and spinotrapezius ‐ have been adapted for specializations relating to arboreal, aquatic, fossorial, cursorial, and flying locomotor patterns. Yet despite the diversity of forms, features persist which may give us clues about evolutionary history. This is especially important for the controversial clade Afrotheria, which is based on genetic data but has no unifying skeletal or dental characters and a scanty fossil record. My research indicates that there are several characters of the trapezius complex which unite some members of the Afrotheria. For example, the sternomastoideus originates from the mandible, the clavotrapezius inserts on the sternum rather than the clavicle and has a slip to the pinna, and the acromiotrapezius fuses with the scapular head of deltoideus. There are also several characters of the trapezius complex which some members of Afrotheria share with other mammals. For example, some members of Eulipotyphla also have a bifid acromiotrapezius and a lumbar origin of the spinotrapezius, and the aclavicular Afrotherians share with Artiodactyla the fusion of clavotrapezius and cleidomastoideus with the clavicular head of deltoideus.

Grant Funding Source: none
Texas Tech System (TTS) for student peer teaching in the anatomy laboratory: subjective and objective assessment
Authors: Lee Vaughan , Vidic Branislav , Schneider Brandt , Hewetson Anthony
Published: June 2021

In a continuing effort to preserve viability of laboratory dissection and, at the same time, accommodate a reduced number of faculty instructors, the TTS has been modified and expanded. The purpose of this report is to disclose how the principals of the TTS have evolved to motivate students for independent group learning through peer teaching in the laboratory. A majority of dissection sessions (53%) were assigned to paired students (designated instructors, DIs) per group of five to direct the technical (dissection) and conceptual (comprehension and application) processes of the assignment. Each DI was assigned through alternating rotation an equal load of instruction throughout the course. DIs were provided: TTUHSC dissection manual with integrated videos, prosections, digital images, and faculty previews. Institutional examination results utilizing TTS sessions were comparable to the results with traditional faculty assisted teaching. Assessment of peer teaching was expanded from subjective measures to include objective practical quizzes. The new assessment enhanced team work and increased student satisfaction. The TTS reduced faculty number and time requirements while maintaining comprehension of laboratory material through self‐learning, differential reasoning, and team work. This project was funded by the Department of Cell Biology and Biochemistry, TTUHSC.
O‐GlcNAc cycling and Epigenetics
Authors: Hanover John A , Wang Peng , Ghosh Salil , Keembiyehetty Chithra , Comly Marcy , Bond Michelle , Krause Michael , Love Dona
Published: June 2021

The nutrient‐sensing hexosamine signaling pathway terminating in O‐GlcNAc cycling is emerging as a key epigenetic regulator of gene expression in mammals. Mounting evidence suggests that OGlcNAc cycling sits atop a robust regulatory network maintaining higher‐order chromatin structure and epigenetic memory. OGlcNAc cycling occurs on many cellular targets associated with signaling, morphogenesis and transcriptional regulation. In addition, it has now been added to the multifaceted ‘histone‐code’. In model systems ranging from C. elegans to man, O‐GlcNAc cycling has been shown play an important role in modulating developmental plasticity. We have focused on the influence of O‐GlcNAc cycling upon the molecular modifications of the RNA Polymerase II carboxyl‐terminal tail. In C. elegans, O‐GlcNAc marks the promoters of over 800 developmental, metabolic, and stress‐related genes. In response to starvation or feeding, levels of O‐GlcNAc at promoters remain nearly constant due to dynamic cycling mediated by the transferase OGT‐1 and the O‐GlcNAcase OGA‐1. Blocked O‐GlcNAc cycling leads to a striking nutrient dependent accumulation of O‐GlcNAc on RNA Pol II. O‐GlcNAc cycling mutants also show an exaggerated, nutrient‐responsive redistribution of promoter‐proximal RNA Pol II isoforms and extensive transcriptional deregulation. These studies in the nematode have been extended to the fly and vertebrate RNA Pol II CTD. Our findings suggest a complex interplay between the O‐GlcNAc modification at promoters and the kinase dependent ‘CTD‐code’ regulating RNA Pol II dynamics. Nutrient‐responsive O‐GlcNAc cycling may buffer the transcriptional apparatus from dramatic swings in nutrient availability by sustaining a 5′ pool of RNA Pol II poised for rapid reactivation to meet metabolic and developmental fate decisions. These findings have important implications for the inheritance of susceptibility to the diseases of aging including diabetes, cancer and neurodegeneration.
Regulation of the calpain and ubiquitin proteasome system in a canine model of muscular dystrophy with myostatin inhibition
Authors: Cotten Steven W. , Wadosky Kristine M. , Bogan Dan , Kornegay Joe N. , Willis Monte S.
Published: June 2021

Recent studies have suggested that inhibiting the protein myostatin, a negative regulator of skeletal muscle mass, may improve outcomes in patients with Duchenne Muscular Dystrophy (DMD) by enhancing muscle mass. We tested this hypothesis by crossing the golden retriever muscular dystrophy (GRMD) dog with the whippet dog having a homozygous mutation for the myostatin‐null allele (Mstn −/−, the so‐called bully whippet). Surprisingly, individuals with affected muscular dystrophy and decreased myostatin demonstrated an accelerated physical decline compared to related GRMD affected dogs with full myostatin. To examine mechanisms responsible for this accelerated decline, we measured novel ubiquitin ligases, proteasome subunits, and calpain 1/2 by real time PCR in the cranial sartorius and vastus lateralis muscles to determine the effects on the ubiquitin proteasome in wild type, GRMD, and GRMD/Myostatin +/− dogs (N=10 total). While affected GRMD dogs had decreases in the ubiquitin ligases MuRF1, Atrogin‐1, RNF25, RNF11, and CHIP, affected GRMD dogs with decreased myostatin exhibited enhanced levels of MuRF1 and Atrogin‐1. These studies suggest a role of myostatin in the disease progression of GRMD, which may involve key components of the ubiquitin proteasome system involved in the turnover of sarcomere proteins in the skeletal muscle.
In Vivo moduation of murine polcystic kidney (PKD) pathology by a single mutated inv gene
Authors: Gattone Vincent H. , Carr Alexander J. , Bacallao Robert L.
Published: June 2021

Significant variability between progression of PKD in two affected by the same mutation suggests modifier gene influence. PKD murine modifier gene loci were associated with chromosomal locations that harbor other PKD genes. Polymorphisms of a single mutant gene for an autosomal recessive(AR) disease could influence severity of pathology caused by a different locus. Interactions between gene products causing nephronophthisis (NPHP) include inv(nphp2) with nphp3 and nphp9. The inv locus plays a role in normal ciliogenesis, critical for PKD. We hypothesized that a mutant copy of inv would be enough to alter nphp3‐induced PKD, in vivo. FVB‐inv mutant mice were crossed with CD1‐pcy(nphp3) mice and pcy cystic pathology compared between those with inv/+ or +/+ genotype. Cystic pcy/pcy renal pathology was accelerated by the presence of the inv/+ genotype. The cpk gene is also known to interact with nphp genes and a modifier loci for cpk severity is on chromosome 4 roughly where inv gene resides. Unlike the crossing results with pcy, inv does not appear to influence the severity of cpk/cpk ARPKD (neither in kidney nor biliary pathology). In vivo studies are required to assess modifying gene capability of inv and other critical PKD associated genes.
Linking tumor associated macrophages, inflammation, and intestinal tumorigenesis: Role of MCP‐1
Authors: Murphy E Angela , Davis J Mark , McClellan Jamie L , Steiner Jennifer L , Jung David , Carmichael Martin D , Carson James A , Pena Maria M , Berger Franklin G
Published: June 2021

Monocyte chemoattractant protein 1 (MCP‐1) is thought to be the most important chemokine for the recruitment of macrophages (MΦs) to the tumor microenvironment. We examined the role of MCP‐1 on tumor associated MΦs, inflammation, and intestinal tumorigenesis in a genetic mouse model of colon cancer. Male ApcMin/+, ApcMin/+/MCP‐1−/− or wild type mice (n=10/group) were sacrificed at 18 wks of age and intestines were analyzed for polyp burden, apoptosis, β‐catenin expression, MΦ number & phenotype, and inflammation. MCP‐1−/− decreased overall polyp number by 25% and large polyp number by 45% (P<0.05). This was consistent with an increase in apoptotic cells (P<0.05). Similarly, the number of β‐catenin positive polyps was reduced in ApcMin/+/MCP‐1−/− mice (P<0.05). MCP‐1 deficient mice showed decreased F4/80 positive cells in the polyp tissue (P=0.06) and surrounding intestinal tissue (P<0.05), and there was a decrease in CD206 mRNA expression (P<0.05). Further, MCP‐1−/− offset the increased mRNA expression of IL‐1β & IL‐6 in intestinal tissue and IL‐1β & TNF‐α in polyp tissue (P<0.05) and prevented the decrease in SOCS1 expression (P<0.05). Circulating IL‐6 levels were also decreased in MCP‐1−/− mice (P<0.05). We demonstrate that MCP‐1 is an important mediator of tumor growth and cancer‐induced inflammation and may serve as an important biomarker and/or therapeutic target in colon cancer.