bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of LDL receptor in the uptake of beta‐carotene by murine placenta and the embryo
Authors: Shete Varsha , Costabile Brianna , Quadro Loredana
Published: June 2021

Beta‐carotene (bC) is the most abundant precursor of vitamin A in the human diet. Vitamin A derived retinoids are essential for normal embryonic development. Uptake of intact bC followed by local de novo biosynthesis of retinoids by cleavage of bC can be an important source of retinoids during embryonic development. Intact bC is circulated in association with various lipoprotein particles, especially apoE/apoB containing lipoproteins such as chylomicrons, VLDL and LDL.

Potential role of LDL receptor (LDLr) in the uptake of supplemented bC by maternal and developing tissues was investigated. LDLr knockout female mice were mated with LDLr heterozygous males and vice versa to obtain embryos of the same litter that are either heterozygous or lack LDLr altogether. All mice were maintained on a regular chow diet throughout the lifespan. At 13.5 days of gestation, dams were intraperitoneally supplemented with a physiological dose of 20–50 μg/g body weight of bC. Maternal serum, liver, placenta and embryos were collected after 24 hours and analyzed by HPLC and qPCR.

Hepatic bC levels were reduced in LDLr knockout dams. Placenta of heterozygous embryos had higher bC levels as compared to the knockout placenta, an effect observed only in case of LDLr knockout dams. Maternal and embryonic genotype did not have an effect on bC levels in embryos. Preliminary qPCR analysis of placenta revealed that both bC supplementation and LDLr genotype of the placenta had an effect on the mRNA expression of genes involved in lipid metabolism. These results suggest that LDLr mediates uptake of bC 1) in the liver and 2) in the placenta of dams lacking LDLr. In addition to this, the data suggested that a single dose of bC at mid‐gestation may alter genes involved in the lipid metabolism in the placenta and the embryo.

Grant Funding Source: NIH
Exploiting the synthetic capacity of microbes for the production of novel value‐added biochemicals
Authors: Jones Prather Kristala L.
Published: June 2021

Growing interest in a biomass‐based economy has led to new efforts to construct and improve microorganisms capable of producing both novel biofuels and value‐added compounds. The potential for biological conversion of feedstocks to bulk chemicals is enhanced by the availability of tools and techniques from the established discipline of Metabolic Engineering, which has enjoyed tremendous successes in the development of highly productive microorganisms for a variety of products of interest. We can also gain insights from Biocatalysis, where the choice of enzymes to mediate biotransformation of chemical substrates is based largely on consideration of the required functional group conversion without being limited by prior evidence of transformation of the full structure, and apply new tools developed under the umbrella of Synthetic Biology. Our group is interested in applying principles from each of these intellectual arenas towards the design and construction of novel biosynthetic pathways for specified target compounds. Using this approach we have developed a novel pathway towards 3,4‐dihydroxybutyric acid and its related lactone 3‐hydroxy‐γ‐butyrolactone, a so‐called “top value‐added” compound with no identified natural source. Results will also be presented on the flexibility of the platform pathway for the production of other, structurally distinct 3‐hydroxyalkanoic acids.
Effect of dietary mannoheptulose on whole body glucose and energy metabolism in adult neutered male Labrador Retrievers
Authors: McKnight Leslie L , Flickinger Elizabeth A , Davenport Gary , France James , Shoveller Anna K
Published: June 2021

The objective of this study was to determine the effect of dietary mannoheptulose (MH) on glucose and energy metabolism in adult male neutered Labrador Retrievers (N = 6, 5.4 y). This study was designed as a cross‐over with each dog receiving both dietary treatments, control (CON) and MH (200 mg/kg), in random order. Fasting and post‐prandial respiratory quotient (RQ) and energy expenditure (EE) were determined by indirect calorimetry (d 16). Glucose kinetics were assessed during fasting and repeated meal feeding using indirect calorimetry and a primed continuous infusion of U‐13C‐glucose (d 18). A fasting biceps femoris muscle sample was obtained (d 21/22) to determine protein content of phosphorylated and total adenosine monophosphate‐activated protein kinase (AMPK) and acetyl CoA carboxylase (ACC). Diet did not affect plasma glucose, insulin, FFA or glucose turnover during fasting or repeated meal feeding. There were no diet‐related differences in glucose oxidation or energy metabolism in the fed state. However, there were trends for MH to increase RQ (p=0.14) and glucose oxidation (p=0.16) and to lower the ratio of phosphorylated to total AMPK protein content (P=0.08) during fasting. These findings suggest daily MH feeding may affect carbohydrate utilization during fasting.

Grant Funding Source: Procter and Gamble Pet Care
Berry anthocyanin fractions repress pro‐inflammatory gene expression and secretion by inhibiting nuclear translocation of NF‐κB in RAW 264.7 macrophages
Authors: Lee Sang Gil , Kim Bohkyung , Yang Yue , Park Youngki , Koo Sung I. , Chun Ock K. , Lee Jiyoung
Published: June 2021

Antioxidant capacity of anthocyanins is known to be associated with their anti‐inflammatory properties. We examined the association between antioxidant capacity of three berry anthocyanin fractions and their anti‐inflammatory potency in lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophages. The anthocyanin fractions of three berries contain different major anthocyanin compounds: blueberry (BB, 16% malvidin‐3‐glucoside), blackberry (BK, 98% cyanidin‐3‐glucoside) and blackcurrant (BC, 44% delphinidin‐3‐rutinoside). BK showed a significantly higher total antioxidant capacity compared with BB and BC in a non‐cell based assay. At 10 and 20 μg/mL concentrations, all of the three berry anthocyanin fractions significantly decreased interleukin‐1β and tumor necrosis factor α (TNFα) mRNA levels in LPS‐treated macrophages to a similar degree. TNFα secretion to cell culture medium was also significantly decreased by the berry fractions although BK showed the least potency. Increased nuclear factor κB (NF‐κB) p65 translocation to the nucleus by LPS was markedly attenuated by treatment with BB, BK and BC. LPS did not increase cellular ROS levels. In conclusion, anti‐inflammatory functions of berry anthocyanins are primarily attributed to their inhibitory role in NF‐κB nuclear translocation with a minimal contribution from their antioxidant capacity in LPS‐stimulated RAW 264.7 macrophages.
Effects of Elderberry Juice on Citrobacter rodentium‐induced Host Responses in C57BL/6J Mice
Authors: Lu Chi‐Hua , Browning Jimmy D , Spagnoli Sean T , Lei Wei , Eichen Peggy A , Fritsche Kevin L
Published: June 2021

Introduction

C. rodentium (CR) is a murine pathogen that shares characteristics with human pathogenic E. coli. Elderberry juice (EJ) has been shown to possess anti‐inflammatory activities. The goal of this study was to evaluate EJ's impact on host immune responses to CR infection.

Experimental Design

Animals (n=40) were randomly assigned into four treatment groups: 1) Uninfected: CR (−), AIN93G diet with no EJ (0% EJ diet); 2) CR (+), 0% EJ diet; 3) CR (+), 0.4% EJ diet; 4) CR (+), 2% EJ diet. CR was given by a single oral dose (10^9 CFU) on day 0, and animals were sacrificed on day 10.

Results

Bodyweight and fecal CR load were similar among groups post‐challenge. H&E staining showed that CR increased colon mucosal height (50%), and pathology scores of inflammation (2‐fold), epithelial defects (2.5‐fold) and hyperplasia (2‐fold). Quantitative PCR showed that CR increased TNF‐alpha (3‐fold), IFN‐gamma (2‐fold) and IL‐17 (to detectable) expression in colon. However, when comparing all infected groups, no such differences were observed by EJ treatment, except for higher IL‐10 expression in colon (2‐fold, p<0.05). Systemic anti‐oxidative status was also evaluated by ORAC and TBARS in plasma and liver, and no significant differences were found among all groups.

Conclusion

Except for elevating colonic IL‐10 mRNA expression, EJ treatment did not significantly alter other inflammatory responses to GI infection by CR.

Grant Funding Source: NIH P50AT006273
Iron stores and home stimulation independently relate to infant development in a rural Indian community with high rates of infant anemia
Authors: Rao Sylvia Fernandez , Black Maureen , Hurley Kristen , Balakrishna Nagalla , Radhakrishna KV , Tilton Nicholas , Reinhart Greg , Harding Kim , Nair K. Madhaven
Published: June 2021

Objective

Examine how iron stores and home stimulation relate to infant development in rural India.

Methods

Enrolled 497 infants age 6–12 mo. Calculated weight‐age, length‐age, & weight‐length z‐scores (WAZ, HAZ, WHZ). Assessed home stimulation (HOME Inventory), motor, language & visual reception (Mullen Scales of Early Learning) & socioemotional (Brief Infant‐Toddler Socio‐Emotional Assessment). Associations of iron stores & home stimulation with development estimated by analyses of variance, adjusted by child age, WAZ, HAZ, WHZ, hemoglobin (Hb), B12, C reactive protein, maternal education & household assets.

Results

71% had adequate iron stores (ferritin>;12 μg/L), 66% were anemic (Hb<11 g/dL), 20% stunted (HAZ<‐2) & 19% underweight (WAZ<‐2). Ferritin was positively associated with fine motor (p=.039), receptive language (p=.045), & socioemotional (p=.023), but not gross motor, expressive language, or visual reception. Home stimulation was positively associated with receptive (p=.025) & expressive language (p=.002), visual reception (p<.001), & socio‐emotional (p<.001), but not gross/fine motor.

Conclusion

Iron stores & home stimulation are independently associated with infant development, supporting evaluation of an integrated micronutrient‐infant development trial.

Support: Mathile Institute & Micronutrient Initiative
Influence of dietary saturated fat content on adiposity, macrophage behavior, inflammation, and metabolism: composition matters
Authors: Enos Reilly , Davis J. Mark , Velazquez Kandy , McClellan Jamie , Carnevale Kevin , Murphy E. Angela
Published: June 2021

We examined the effects of three high‐fat diets differing in the percentage of total calories from saturated fat (SF) (6%, 12%, and 24%), but identical in total fat (40%), on body composition, macrophage behavior, inflammation, and metabolic dysfunction in mice. Diets were administered for 16 weeks. Body composition and metabolism (glucose, insulin, triglycerides, LDL‐C, HDL‐C, total cholesterol) were examined monthly. Adipose tissue (AT) expression of marker genes for M1 and M2 macrophages and inflammatory mediators (TLR‐2, TLR‐4, MCP‐1, TNF‐α, IL‐6, IL‐10, SOCS1, IFN‐γ) was measured along with activation of NFκB, JNK and p38‐MAPK. AT macrophage infiltration was examined using immunohistochemistry. Circulating MCP‐1, IL‐6, adiponectin, and leptin were also measured. SF content, independent of total fat, can profoundly affect adiposity, macrophage behavior, inflammation, and metabolic dysfunction. In general, the 12%‐SF diet, most closely mimicking the standard American diet, led to the greatest adiposity, macrophage infiltration, and insulin resistance (IR), whereas the 6%‐SF and 24%‐SF diets produced lower levels of these variables with the 24%‐SF diet resulting in the least degree of IR and the highest TC/HDL‐C ratio. Macrophage behavior, inflammation and IR following HFDs are heavily influenced by dietary SF content, however, these responses are not necessarily proportional to the SF%. This work was supported by an ASPIRE (Advanced Support Programs for Innovative Research Excellence) grant from the University of South Carolina to E.A.M.

Grant Funding Source: ASPIRE
iCook: Development of web‐based component of a 4‐H cooking intervention
Authors: Colby Sarah E , Olfert Melissa , Mathews Douglas , Kattelmann Kendra K , Franzen‐Castle Lisa , White Adrienne
Published: June 2021

A web‐based component was developed to support a cooking intervention implemented through Extension programming in 5 states. The intervention was pilot‐tested with dyads (n=61) of youth (9–10 years old) and adults. Six lessons included content on family mealtime and communication, physical activity, and goal setting. Youth received cameras and were asked to create videos on the lessons for the website which was also designed for interaction with other participants and goal setting. Based on process evaluation, 75% of youth reported making videos but only 48% reported posting videos. Barriers to posting videos included slow Internet speed at home, lack of time and forgetfulness. While 65% of youth reported intent to set goals to increase fruits, vegetables and physical activity, goal setting on the website was completed only 38 times. Youth need support built into program structure to follow through with intentions. Although Internet access has been identified as increasingly available, the barrier of slow speed Internet access, i.e., upload speed, limits posting capabilities and is important to consider when developing online interactive at‐home activities. Feedback will be used to modify online components. Funding Source: USDA/NIFA/AFRI, Integrated Proposal for Childhood Obesity Prevention Challenge Area #2012–68001‐19605 and state experiment stations.
Restricted diet benefits body composition but deteriorates bone remodeling in middle‐aged obese female rats
Authors: Shen Chwan‐Li , Dagda Raul Y , Chyu Ming‐Chien
Published: June 2021

We investigated the effects of a high‐fat diet (HFD, 40% energy from fat) and a restricted diet (RD, 35% energy deficit) on body composition and bone parameters in obese female rats. Twenty 3‐month‐old SD female rats were fed with a HFD at libitum for 4 months, and were then randomly assigned to either a HFD or a RD for another 4 months. Another 10 3‐month‐old SD female rats in a low‐fat diet (LFD, 10% energy from fat) for 8 months served as the control group. Body composition was measured by bioimpedance spectroscopy. Bone efficacy was evaluated by dual‐energy absorptiometry, histomorphometry, and 3‐point bending test. Serum leptin, adiponectin and insulin‐like growth factor‐I (IGF‐I) were determined. Relative to the HFD group, the RD group demonstrated (i) lower body weight, % fat mass, leptin, and IGF‐I, and higher % fat‐free mass, and adiponectin, and (ii) suppressed bone formation and increased bone resorption, resulting in decreased trabecular and cortical volume, bone mineral density and strength. Relative to the LFD group, the RD group demonstrated similar body composition and serum biomarker results, higher bone formation and resorption, and lower bone mineral density and strength. Our data suggest that although RD benefits body composition, it is detrimental to bone remodeling in obese female rats.
Predictors of body mass index of Colombian 5 to 18 year old children
Authors: Gonzalez‐Casanova Ines , Sarmiento Olga Lucia , Pratt Michael , Stein Aryeh
Published: June 2021

We examined the role of municipality, household, and individual factors on body mass index (BMI) among children 5 to 18 years measured in the Colombian National Nutrition Survey of 2005 (n=7,802) and 2010 (n=10,302). We used multilevel models with municipality as level‐3 (park density and GINI inequality index as predictors), household as level‐2, child as level‐1, and BMI z‐score (BMIZ) relative to the WHO reference population as outcome. Municipality explained 3% of variability in BMIZ in 2005 and 2% in 2010. Park density (0.03 per increase in park z‐score, p<0.05) and inequality (0.05 per unit increase in GINI z‐score, p<0.01) were positively associated with BMIZ in 2010. Household level explained 40% of variability in 2005 and 31% in 2010. Wealth (0.07 and 0.12 per quintile increase) and living in an urban section of the municipality (0.24 and 0.12 vs. rural, p<0.05) were positively associated with BMIZ in both 2005 and 2010. Being part of an extended family was positively associated (0.08 vs. nuclear, p<0.01) and family size was inversely associated (−0.05 per member increase, p<0.01) with BMIZ in 2005. Age was inversely associated and being a girl was positively associated with BMIZ in both surveys. The 2005 and 2010 models differed. The positive relationship with park density and inequality is counterintuitive. The association between wealth and BMIZ differs from that observed in developed country settings.
iCook: Use of an online process evaluation to facilitate quality control of a 5‐State 4‐H program.
Authors: Mathews Douglas R , Yerxa Kathryn , Franzen‐Castle Lisa , Krehbiel Michelle , Colby Sarah , Meade Randa , Kattelmann Kendra , Kabala Celine , Olfert Melissa , Flanagan Sue , White Adrienne A
Published: June 2021

An online process evaluation tool was used to enhance quality control during a 5‐state childhood obesity prevention pilot study. Family dyads (n=61; youth aged 9–10 and their primary adult food preparer) participated in 6‐biweekly lessons to promote four priority areas (culinary skills; family meals; physical activity; food and activity goal setting). After each lesson, dyads completed online surveys, which were reviewed by researchers and lesson leaders for program improvement. Based on parent reporting on priority area goals, from earlier to later in the program, increased percentages were seen for number of biweekly family meals (82% to 88%), cooking with their children (35% to 57%) and decreased percentage for being active as a family (49% to 39%). Over the program, youth identified cooking (75%) and tasting (62%) as learning experiences compared to practicing communication at mealtime (29%). Online process evaluation facilitated researchers’ monitoring and responsiveness to multistate study implementation. Researchers were able to identify the translation of lesson priorities to the home environment and make changes for program improvement during pilot testing to improve the probability of positive intervention outcomes. Funding Source: USDA/NIFA/AFRI, Integrated Proposal for Childhood Obesity Prevention Challenge Area #2012–68001‐19605 and state experiment stations.
Beneficial Effects of Green Tea Consumption in Parkinson's Disease Patients
Authors: Chen Dan , Lyons Kelly , Pahwa Rajesh , Zhou Ying , Reddy Manju
Published: June 2021

Antioxidants may offer protection by reducing oxidative stress associated with Parkinson's disease (PD). The objective of this study was to assess the beneficial effect of green tea (3 cups) consumption for 3 months in enhancing antioxidant status, reducing oxidative damage, and improving quality of life of PD patients. Fourteen subjects (51–79 y) who were within first five years of PD onset, on stable medication and not regular green tea consumers were recruited. PD rating scales, antioxidant enzymes, oxidative stress markers, and iron status were assessed at baseline and after 3 months. Among the PD rating scales, we found an increase in Mini‐Mental State Examination (p<0.05) and a decrease in Unified Parkinson's Disease Rating Scale (UPDRS) motor scores (p<0.05) after the intervention, suggesting an improvement in cognitive and PD symptoms. Compared to baseline, activity of catalase and superoxide dismutase were increased significantly (28%, p<0.05 and 37%, p=0.0025, respectively) but not glutathione peroxidase. A significant, 52% reduction in both lipid peroxidation (p<0.0001) and protein carbonyls (p=0.0004) was found after intervention. No changes in iron status were found. In conclusion, green tea consumption may be beneficial in reducing oxidative stress in PD patients and may offer protection against progression of the disease.
Improving Head Start's system of communicating children's weight status and BMI scores to caregivers
Authors: DuBois Matthew R , Hoffman Jessica , Carter Sonia , Bottino Clement , Nethersole Shari , Cox Colin , Sceppa Carmen , Wirth Cathy
Published: June 2021

Members of a community‐academic partnership collaborated to strengthen Head Start‐family communication regarding children's weight status. Head Start serves low‐income preschoolers and is mandated to measure children's Body Mass Index (BMI). Head Start staff was concerned because 40% of their children are categorized as overweight/obese and caregivers rarely respond to letters reporting BMI data. In spring 2012, academic partners collaborated with Head Start to form a working group to improve BMI communication with caregivers. As compared to baseline, working group participants rated childhood obesity as significantly more important after completing the group process. A task force was then formed to prioritize and implement group recommendations. Two recommendations were implemented in fall 2012; creating a staff training about healthy weight and revising the BMI letter. Following the training, staff felt more prepared to talk to caregivers about BMI and reported the revised BMI letter was easy to understand. Next, focus groups were held at four Head Start programs to evaluate caregiver perceptions of the revised letter. Caregivers reported that the letter was helpful and provided valuable information. Overall, the partnership improved Head Start's capacity to provide caregivers with important health information about their children. Support: Northeastern University, Boston Children's Hospital.
Aging alters global hepatic DNA hydroxymethylation in mice, as determined by a novel LC/MS‐MS method
Authors: Tammen Stephanie A. , Dolnikowski Gregory G. , Ausman Lynne M. , Friso Simonetta , Choi Sang‐Woon
Published: June 2021

Aging is associated with changes in global DNA methylation (mC), but changes in DNA hydroxymethylation (hmC) are not established due to a relative lack of methods available to measure hmC. Both modifications can affect gene expression and may be important in aging. We have developed a method to measure 5′‐methyl‐2′‐deoxycytidine and 5′‐hydroxymethyl‐2′‐deoxycytidine in DNA using liquid chromatography/tandem mass spectrometry (LC/MS‐MS) and used this method to assess global hmC and mC levels in hepatic tissue from young (9 months; n=10) and old (23 months; n=6) C57BL/6 male mice. DNA was enzymatically hydrolyzed and isotopomers [15N3]‐2′‐deoxycytidine and (methyl‐d3, ring‐6‐d1)‐5‐methyl‐2′‐deoxycytidine were added as internal standards. The mixture of DNA hydrolysates and internal standards were separated through LC, and nucleosides of interest were detected by MS‐MS in positive ion mode using multiple reaction monitoring. Global mC and hmC levels were calculated as a percentage of total deoxycytidine residues in genomic DNA. Liver tissue from old mice had significantly higher global hmC relative to young mice (0.32±0.02% vs 0.24±0.01%; p=0.02), while mC levels did not change. Using this new method we have found that aging is associated with an increase in global DNA hmC, a finding that may be useful in understanding the physiological mechanisms behind aging.

Grant Funding Source: NIH grant R01 AG025834
Low‐glycemic index diets prevent type 2 diabetes in the Nile rat
Authors: Bolsinger Julia , Pronczuk Andrzej , Hayes KC
Published: June 2021

The Nile rat (NR) is a novel model for type 2 diabetes (T2DM) and metabolic syndrome sensitive to metabolic stress from simple carbohydrates (CHO). Three experiments assessed the effects of a low‐glycemic index CHO (green lentils, GL) on T2DM. Expt 1 compared a GL diet directly with a semipurified hiCHO diet having identical %En from CHO:fat:protein (70:10:20). Expt 2 assessed reversibility of hyperglycemia by comparing a modCHO‐GL diet (45:25:30) to diabetogenic semipurified hiCHO (70:10:20) or lab chow (#5020) diets in 3 separate studies. Expt 3 examined young Nile rats “primed” from weaning for 7wks with the modCHO‐GL diet (45:25:30) before being challenged with stressful semipurified hiCHO (70:10:20) or modCHO (40:40:20) diets based on glucose and starch. 1. Compared to the hiCHO semipurified diet with the same macronutrient composition, the GL diet induced significantly less diabetes. 2. GL substantially reversed hyperglycemia, especially in young (<4mo) diabetic rats. 3.”Priming” rats with GL prior to hiCHO resulted in less severe diabetes compared to previous studies without priming. Positive influences on other metabolic parameters such as liver and kidney weight and plasma lipids also were observed. Since the NR mimics T2DM in humans, GL or components therein may represent a potentially useful adjunct for T2DM control in humans.

Funding source: Foster Biomed Res Lab and the Malaysian Palm Oil Board
Botanicals inhibit castrate‐resistant prostate cancer via mechanisms involving AMPK, inhibition of lipogenesis, mTOR‐mediated proliferation, and histone modification
Authors: Zhao Yi , Whelan Jay
Published: June 2021

Reprogramming of metabolic pathways and abnormal acetylation of DNA are two common features of cancer cells, including prostate cancer (PCa). Our results show that polyherbal mixtures of botanicals, like Zyflamend®, can modify lipogenesis in CWR22Rv1 cells (a castrate‐resistant human PCa cell line) by enhancing the activity of 5'adenosine monophosphate (AMP)‐ activated protein kinase (AMPK), a potential tumor suppressor protein that functions as a central energy sensor within the cell. By increasing the phosphorylation of AMPK, Zyflamend® decreases de novo lipid biosynthesis by decreasing the activity of acetyl‐CoA carboxylase (ACC) and the expression of FASN and SREBP1c. Also via AMPK, Zyflamend® inhibits the activity of mTORC1 complex by directly phosphorylating Raptor, decreasing the activity of P70 (S6K, the marker for protein synthesis). All of these effects can be mimicked by treatment of AICAR (activator of AMPK). Furthermore, Zyflamend® increases the acetylation of DNA via mechanisms that appear to involve ATP‐citrate lyase (ACLY). Thus, our results suggest that botanicals, via AMPK, inhibit castrate‐resistant prostate cancer via mechanisms that involve lipid metabolism that may reshuttle acetyl CoAs for histone acetylation, and inhibition of mTOR mediated cell proliferation.

Grant Funding Source: This research was supported in part by Tennessee Agricultural Experiment Station
Molecular characteristics determining entry into or exclusion from the endothelial cell (EC) lateral border recycling compartment (LBRC)
Authors: Feng Gong , Muller William A.
Published: June 2021

The LBRC is a recently discovered membrane compartment in EC. It contains molecules critical for leukocyte transmigration, PECAM, CD99, and JAM‐A, but excludes other junction components like VE‐cadherin. Since a mutation in the cytoplasmic tail (C‐tail) of PECAM prevented localization to the LBRC, we first determined whether the C‐tail of PECAM could bring a molecule not expressed on EC into the LBRC. A chimera with the extracellular domain (ECD) of Tac and C‐tail of PECAM localized to junctions, entered the LBRC, and recycled like PECAM. However, the control protein, Tac, entered the LBRC too, even though it was expressed diffusely. This suggested that entrance to the LBRC might be default and VE‐cadherin is selectively excluded. We constructed a VE‐cadherin/PECAM (CP) chimera with the ECD of VE‐cadherin and C‐tail of PECAM. We found CP went to junctions but was excluded from the LBRC, like VE‐cadherin. These findings suggested ECD excluded VE‐cadherin and CP from the LBRC. We then generated truncated VE‐cadherin and CP with deletion of the homophilic interaction domain EC1 or its specific motif RVDAE. Both VE‐cadherin and CP chimera lacking RVDAE motif or EC1 domain DID enter the LBRC. This suggests that homophilic interactions of VE‐cadherin with other VE‐cadherin molecules stabilize it at cell borders and prevent entry into the LBRC. Supported by AHA Postdoctoral Fellowship to GF and R01 HL046849 to WAM
Pressure‐Overload Hypertrophy of the Developing Heart Reveals Activation of Divergent Gene and Protein Pathways in the Left and Right Ventricular Myocardium
Authors: Friehs Ingeborg , Cowan Douglas B , Choi Yeong‐Hoon , Black Kendra M , Barnett Reanne , Del Nido Pedro J , Levitsky Sidney , McCully James D
Published: June 2021

Right (RV) and left ventricular (LV) myocardium differ in their pathophysiological response to pressure‐overload hypertrophy (POH). Herein we use microarray and proteomic analyses to identify pathways modulated in LV‐ and RV‐ POH. Newborn New Zealand White rabbits underwent banding of the descending thoracic aorta (LV‐AOB; n=6) or banding of the pulmonary artery (RV‐PAB; n=6). Controls (SC; n=6 each) were sham‐manipulated. Following 4 (LV‐AOB) and 6 weeks (RV‐PAB) recovery, the hearts were removed and matched RNA and protein samples were isolated for microarray and proteomic analysis as compared to SC. Our results demonstrate there was increased transcript expression levels for mitochondria energy pathways, actin, hypoxia, calcium and protein kinase‐A signalling and increased protein expression levels for cellular macromolecular complex assembly and oxidative phosphorylation in LV‐AOB. In RV‐PAB there was increased transcript expression levels for cardiac oxidative phosphorylation and increased protein expression levels for cardiac muscle tissue development and calcium handling. Our results demonstrate that the mitochondrion plays an important but varied role to support divergent compensatory pathways in LV‐ and RV‐POH. These data provide new insight into the biological basis of LV‐ and RV‐POH during compensation and identify novel therapeutic pathways for intervention prior to failure.
Acidic Sphingomyelinase Inhibition Limits CCl4 Induced Hepatic Fibrosis
Authors: Quillin Ralph Cutler , Wilson Gregory , Nojima Hiroyuki , Wang Jiang , Schuster Rebecca , Blanchard John , Edwards Michael , Gulbins Erich , Lentsch Alex
Published: June 2021

Existing evidence suggests that acidic sphingomyelinase (ASMase) controls hepatic stellate cell activation and may contribute to fibrosis. The objective of this study was to determine if the pharmacologic inhibition of ASMase limits CCl4 induced hepatic fibrosis. C57BL/6 mice were given intraperitoneal injections of CCl4 for 8 weeks to induce fibrosis. Some animals received amitriptyline in their drinking water to inhibit ASMase activity. Liver sections were examined and fibrosis stage (0–4) was scored. Treatment with CCl4 generated significant hepatic fibrosis after week 4 (3.0 ± 0 vs. 1.2 ± 0.3) and week 6 (3.6 ± 0.2 vs. 1.3 ± 0.2) when compared to vehicle. Hepatic cirrhosis (stage 4 fibrosis) was induced after 8 weeks of CCl4 treatment compared to vehicle (3.83 ± 0.41 vs. 0.67 ± 1.0, p<0.05). Inhibition of ASMase caused a significant decrease in hepatic collagen deposition at week 4 (2 ± 0), week 6 (2 ± 0) and week 8 (2.5 ± 0.3) when compared to CCl4 treatment (p<0.05 for each). These data suggest that inhibition of ASMase may be a therapeutic target in hepatic fibrotic disorders and may prevent the progression to cirrhosis.
Using cadaveric material to impact translational research: From anatomy lab to the operating room.
Authors: Lambert H. Wayne , Clarkson Mackenzie J. , Fox Jacob N. , Kennedy Patrick M. , Wisco Jonathan J.
Published: June 2021

The goal of translational research is to enable biomedical research discoveries to go from the “bench to the bedside” to impact health outcomes and improve patient care. Through collaborations with clinical research faculty, an anatomist can actively engage in translational research and increase their research profile within their home institution. This presentation will highlight ways to create clinical research partnerships, ongoing and recently completed translational research projects, resulting scholarly publications, and efforts to secure extramural funding to support these projects. These research collaborations utilized cadaveric material to improve orthopedic surgery outcomes, to document extra‐cardiac neural remodeling following cardiac pathology, and to study age‐related changes in blood vessels which may lead to erectile dysfunction. With the recent funding emphasis of the National Institute of Health (NIH) focused on clinical and translational research, this symposium is timely and aims to inform anatomists and researchers of the necessary steps to enter clinical research and to secure funding.
Mitophagy and Cardioprotection
Authors: Gottlieb Roberta A.
Published: June 2021

Cardiomyocytes rely on autophagy for cellular homeostasis and as a response to nutritional, oxidative, or ischemic stress. Mitophagy, an organelle‐selective form of autophagy, is invoked in the heart in response to cardioprotective interventions such as ischemic preconditioning. An essential regulator of mitophagy is Parkin. Recent work has demonstrated a requirement for Parkin‐dependent mitophagy in the cardioprotection mediated by ischemic preconditioning. Given that autophagy is suppressed in the setting of metabolic syndrome, and given that statins are widely administered to patients with metabolic syndrome, we investigated the effects of statins on autophagy, mitophagy, and cardioprotection. We found that statins induced autophagy through inhibition of mTOR signaling, and induced mitophagy through a Parkin‐dependent pathway. Statin‐mediated cardioprotection was abolished in Parkin‐null mice. Like cholesterol, coenzyme Q10 is a downstream product of HMG‐CoA reductase. Cell‐based studies showed that statin‐dependent mitophagy was abolished by supplementation with coenzyme Q10. Acute supplementation of coenzyme Q10 in mice abolished the cardioprotective effects of statins, raising concern that this widely‐used nutritional supplement could interfere with the cardioprotective benefits of statin therapy.
Principal human milk oligosaccharides utilized by human gut microbiota
Authors: Yu Zhuo‐Teng , Liu Bo , S David
Published: June 2021

Human milk oligosaccharides are poorly digested and absorbed in the small intestine, allowing their passage into the colon, where they influence colonization by the microbiota. The ability of individual major fucosylated and sialylated human milk oligosaccharides to promote growth and physiologic activity of 25 human intestinal microbes was investigated. 2′‐Fucosyllactose, 3‐ Fucosyllactose, and Lactodifucotetraose stimulated most Bifidobacteria spp. and Bacteroides spp. to grow and produce abundant lactate or short chain fatty acids. 3′‐sialyllactose and 6′‐ sialyllactose promoted moderate growth of B. longum JCM7007, 7009, 7010, 7011, 1272, 11347, ATCC15708, B. thetaiotaomicron ATCC29148, and C. leptum ATCC29065; these bacteria produced lactate or short chain fatty acids or both. In contrast, L. rhamnosus ATCC53103, E. facalis ATCC29200, S. epidermidis ATCC12228 and E. coli K12 did not consume the milk oligosaccharides. In summary, specific Bifidobacteria and Bacteroides differentially digest specific individual human milk oligosaccharides, especially the major fucosylated milk oligosaccharides, and these may prove useful for treating dysbiosis of the microbiota and associated inflammatory disorders.

Grant Funding Source: HD013021, AI075563, and Abbott Nutrition
Epigenetic mechanism: silent nucleosomal structures and non‐coding RNAs.
Authors: Kingston Robert E. , Armache Karim‐Jean , Simon Matthew D. , West Jason , Grau Daniel , Garlick Joseph , Davis Chris
Published: June 2021

A paradigm for epigenetic regulation is the stable silencing of genes that are involved in cell fate decisions. This maintained silencing is essential for regulation of genes such as HOX genes in flies and mammals and mating type genes in yeast. Nucleosomes are known to be involved in these silencing mechanisms, both by forming compacted structures and by virtue of their covalent modification. We are studying how nucleosomes might contribute to silenced structures by examining nucleosome occupancy in cells, by using functional biochemical assays, by using electron microscopy, and by using X‐ray crystallography. I will discuss the proteins that can lead to compacted structures, how those structures might function, and our work toward understanding the biological relevance of compacted structures to gene expression. Covalent modification of histones can help to either promote or inhibit formation of these structures and our data address how modifications interact with silencing proteins. Silencing proteins must be targeted in order to function appropriately, and one proposed mechanism for targeting involves non‐coding RNAs. To understand these mechanisms, we have developed a method to map binding sites for ncRNAs in the genome. We are applying these methods to ncRNAs that are involved in epigenetic processes.
Mechanism and function of DHHC S‐Acyltransferases
Authors: Linder Maurine E. , Jennings Benjamin , Lai Jianbin
Published: June 2021

Protein S‐palmitoylation is a reversible posttranslational modification of proteins with fatty acids. In the last five years, improved proteomic methods have increased the number of proteins identified as substrates for palmitoylation from tens to hundreds. Palmitoylation regulates protein membrane interactions, activity, trafficking, and stability and can be constitutive or regulated by signaling inputs. A family of protein acyltransferases or PATs is responsible for modifying proteins with palmitate or other long‐chain fatty acids on the cytoplasmic face of cellular membranes. The signature feature is a DHHC (Asp‐His‐His‐Cys)‐ cysteine rich domain that is the catalytic center of the enzyme. My presentation will focus on what we have learned about enzyme mechanism, regulation, and the determinants of substrate specificity. There are twenty‐three members of the DHHC family in humans. Their biomedical importance is underscored by associations with intellectual disability, Huntington's Disease, and cancer in humans, raising the possibility of DHHC proteins as targets for therapeutic intervention.
Just what the doctor ordered: novel medical student nutritional education through community‐based cooking classes
Authors: Birkhead Andrew Gartner , Loyd Jacob , Leong Benjamin , Joo Esther , Monlezun Dominique , Kay Danielle , Abu‐Shamat Layla , Sarris Leah , Harlan Timothy
Published: June 2021

Adams et al. (2010) demonstrated in a survey of 126 US medical schools that most medical nutrition curriculum hours are taught in a passive lecture format rather than within an active clinical context. This may explain Spencer et. al. (2010) who reported that only 19% of graduating US medical students felt prepared to use nutrition counseling in practice. Until this year Tulane University's medical nutrition curriculum was limited to this model. Tulane's newly created Center for Culinary Medicine combined with the medical student service learning program in an effort to improve nutrition education. Students are required to log 50 hours of community service, which they can now spend learning and teaching principles of cooking and nutrition to patients in ambulatory settings in addition to the traditional curriculum. To date approximately 25% of the Tulane medical student body has participated in nutrition service learning. Students who opted to participate will be compared to students who only had the lecture component by examining perceived competence to counsel patients on a wide range of nutrition topics. Research support: Tulane University Center for Culinary Medicine.

Grant Funding Source: Tulane University Center for Culinary Medicine
Characterization of temporomandibular joint disc in human by histological and biochemical analyses
Authors: Lerma Tanya , Taylor Victor W. , Guttmann Geoffrey D.
Published: June 2021

The temporomandibular joint (TMJ) is characterized by mandibular condyles, capsule, lateral pterygoid, articular surface of the temporal bone, ligaments, and articular disc. For this study the focus is complex articular disc attachments. The main component of the disc is fibrocartilage while its extracellular matrix (ECM) is predominantly composed of collagen type II, sulfated glycosaminoglycan (GAG) and proteoglycans. The purpose is to use histological techniques to investigate components of TMJ disc attachments and quantify constituents of the disc using biochemical assays. Microscopic investigation of attachments provides information to understanding the mechanics of the TMJ disc and an understanding of the relationship between disc and attachments. Six (6) fresh un‐embalmed TMJ disc‐attachment complexes were dissected from willed body donors. TMJ discs were further dissected into five regions and disc attachments were dissected into six regions to emulate a previous study by Willard et al (2011), which used porcine TMJ. Histology and biochemical assays were performed on each region to quantify ECM components. Histological analysis provided information on distribution of components in the complex.

Grant Funding Source: None
Nanoenhanced Hydrogel System with Sustained Release Capabilities for Bone Regeneration
Authors: Karnik Sonali , Hines Kanesha , Mills David
Published: June 2021

Tissue engineering strives to integrate structure and functionality in bioengineering scaffolds or implants. These materials must be biodegradable, immunocompatible, and integrate well with the natural tissue. The bioengineered scaffolds and implants should also be eco‐friendly to manufacture. The aim was to design an implant that enhances inherent bone regeneration y, mimics the natural internal environment and is composed of eco‐friendly materials. We used halloysite nanotubes (HNTs), naturally occurring nanoclay, similar to the kaolin chemically and structurally, and composed of oppositely charged concentric layers of hollow tubules. This structure enables HNTs to acts as a nanocontainer and release bioactive molecules periodically and in a sustained manner. We combined HNTs with alginate hydrogels to release BMPs 2, 4, and 6 and are known to induce osteogenic expression in osteoblasts. We obtained the sustained release of BMPs over a seven‐day period. The amount released was in the range of picograms per milliliter. Osteoblasts encapsulated in these nanoenhanced hydrogels showed enhanced mineralization and ECM secretion as compared with controls over a period of 28 days. Cellular response varied depending on BMP treatment. This nanoenhanced hydrogel system may be a viable solution for in situ bone regeneration for use in degenerative joint disease, small fractures and other bone defects.
Coordination of osteoclast‐osteoblast differentiation by transglutaminase 2
Authors: Nurminskaya Maria V , Lima Florence
Published: June 2021

Enzyme transglutaminase 2 (TG2) promotes maturation of committed osteoblasts in vitro. Nevertheless, TG2 null mice have no skeletal phenotype. Here, we tested the hypothesis that TG2 regulates differentiation in both osteoblasts and osteoclasts with mutually attenuating effects, resulting in the maintenance of status quo between bone formation and resorption. Differentiation of bone marrow‐derived progenitors into osteoblasts and osteoclasts was compared in TG2−/− and wild‐type mice. A significant 25+/− 7% (p<0.001) decrease in the osteoblast colony formation and a dramatic 50+/−15% (p<0.01) decrease in the number of tartrate‐resistant acid phosphatase (TRAP)‐positive colonies was detected in the TG2−/− bone marrow as compared to the wild‐type. Similarly, osteoblast differentiation was delayed in the TG2−/− BMSCs with an 80% reduction in expression of bone sialoprotein and alkaline phosphatase. Further, exogenous purified TG2 accelerated differentiation of the wild‐type progenitors into both osteoblasts and osteoclasts, while the TG2‐specific inhibitor KCC‐009 quenched the TG2‐induced increase in the number of the wildtype osteoclast TRAP‐positive colonies and the number of multinucleated osteoclasts. These data demonstrate a new role for TG2 in coordinating osteoblast‐osteoclast crosstalk and differentiation. This research was supported by NIH grants R03AR057126 and 1R56DK71920.
Inhibition of Notch signalling induces extensive intussusceptive neo‐angiogenesis by recruitment of mononuclear cells
Authors: Djonov Valentin G.
Published: June 2021

Objective

to evaluate the effects of Notch inhibition on the vasculature using chick area vasculosa and MxCre Notch1(lox/lox) mice and to test to which extent this effect is mediated by ephrinB2/EphB4.

Methods and Results

Inhibition of Notch signaling by Y secretase 1 induced significantly intussusceptive angiogenesis (IA) forming large immature capillary plexus. The latter it is characterized by 40% increased vascular density, pericyte detachment, increased transpermeability and recruitment of mononuclear cells to the sites of new formed transluminal tissue pillars (sign of IA). The process was dramatically enhanced when Notch inhibition was combined with the injection of chicken or mouse bone marrow‐derived mononuclear cells. Along with the morphological alterations of IA, we observed down‐regulation of ephrinB2 mRNA levels. Inhibition of ephrinB2 and EphB4 signaling induces pericytes detachment and significantly upregulation of VEGFR1 but without angiogenic response and recruitment of mononuclear cells. Remarkable, the chemotactic factors SDF‐1 and CXCR4 were up‐regulated only due to the Notch inhibition.

Conclusions

Our results indicated that Notch inhibition disturbs vessel stability, induces extravasation of mononuclear cells of bone marrow origin by up‐regulation SDF‐1/CXCR4 chemotactic factors. Recruited cells contributed to formation of transluminal tissue pillars i.e. intussusceptive neo‐angiogenesis, triggering in this way the augmentation of the capillary plexus but without the accompanying vascular maturation and remodeling. .
Chronic ivabradine‐induced heart rate reduction does not modify collagen accumulation in myocardial infarction scar of middle‐aged rats
Authors: Bogatyryov Yevgen , McCooey Daniela , Tomanek Robert J , Dedkov Eduard I
Published: June 2021

Heart rate reduction (HRR) by ivabradine (IVA), a selective inhibitor of the pacemaker If current, for 4 weeks after myocardial infarction (MI) was associated with the reduced collagen accumulation in surviving left ventricular (LV) myocardium. Although this effect is considered to benefit LV performance, the decrease in fibrillar collagen (FC) can diminish a tensile strength of the scar. Therefore, we designed our study to determine whether chronic HRR with IVA affects a structural composition of the MI scar. A large MI (>; 50% of the LV free wall) was induced in 12‐month‐old male Sprague‐Dawley rats by ligation of the left coronary artery and the rats were then assigned in two experimental groups. In the first group, rats were treated with IVA i.p. via osmotic pumps in a dose of 10.5 mg/kg/d for 3 months (MI+IVA), while in the second group, rats received placebo (MI). After three months, the fractional volume of FC was determined in the myocardium of LV free wall and the scar. The scar size and thickness were comparable between MI and MI+IVA rats (63.1±2.7% vs. 59.6±1.8% and 0.93±0.09mm vs. 0.90±0.05mm, respectively). Although, there was a significant reduction of FC in LV free wall of IVA‐treated rats (5.1±0.5% vs. 7.8±0.8%, p≤0.01) the content of FC was comparable in scars of MI and MI+IVA rats (71.8±3.9% vs. 71.7±6.9%, respectively). Our data show that chronic IVA‐induced HRR does not reduce the content of collagen in MI scar.

Grant Funding Source: Internal Award from the NYCOM Office of Research (E.I. Dedkov) and Research Grant from IRI Servier (R.J. Tomanek)