bluuebunny/crossref_metadata_2025 huggingface dataset:
Preschooler's sack lunches do not meet the dietary recommendations for calcium
Authors: Romo-Palafox Maria Jose , Sweitzwer Sara J , Briley Margaret E , Robert-Gray Cindy
Published: June 2021

The objective of this study was to determine the calcium content of preschool sack lunches (n=135) prepared by parents of children age 3–5. Data were collected on 3 nonconsecutive days, analyzed by Food Intake Analysis System (FIAS) to determine the calcium content and compare it to 1/3 of the Dietary Reference Intake (DRI) for children age 3, and 4–5. Portion sizes were evaluated based on Child and Adult Care Feeding Program (CACFP) guidelines. The mean calcium content of sack lunches for three year olds (275.61 ± 38.94 mg) provided more than 1/3 of the DRI (233.33 mg) for their age. Calcium content for four year olds (241.68 ± 34.27 mg) and five year olds (328.89 ± 44.86 mg) did not meet 1/3 of the DRI (333.33 mg). Primary contributing foods to calcium content in sack lunches were cheese (81%), yogurt (42%), fluid milk (21%) cheese spreads (15%), smoothies and other dairy products (13%). The mean serving of milk was 5.6 oz / sack lunch. Parents failed to provide 1/3 of the DRI for calcium in their child's sack lunch for 44% of three year olds, 77% of four year olds, and 50% of five year olds. These findings suggest the need for nutrition education of parents to assure young children receive the necessary calcium from skim milk. Research support National Cancer Institute NCI

Grant Funding Source: R21 CA 123140‐01A
Selling Sprinkles as part of a health products package may reduce fever and diarrhea incidence but not respiratory illness in preschool children in western Kenya
Authors: Grant Frederick Kobina Ebo , Martorell Reynaldo , Ayala-Flores Rafael , Cole Conrad R. , Ramakrishnan Usha , Laird Ruth J. , Patel Minal , Juliao Patricia C. , Quick Rob , Suchdev Parminder S.
Published: June 2021

Background

Supplementation with zinc and vitamin A has been associated with reduced duration and severity of diarrhea and respiratory illness in children.

Objective

We measured the association of community‐based sales of Sprinkles with incidence of diarrhea, fever and cough in children 6–59mo.

Design

This was a cluster‐randomized, longitudinal cohort trial in 60 villages for the first 9mo (July 2007–March 2008), and expansion of access to Sprinkles to both study groups in the second 9mo (July 2008–March 2009). Thus, in addition to the experimental analyses using the first year's data, an as‐treated analysis to compare users and non‐users of Sprinkles across all 60 villages using all available data was carried out. The incidence of diarrhea, cough, and fever were compared between children who used Sprinkles and those who did not.

Results

Of the 1079 children enrolled, 78% (n=847) had data on Sprinkles use. Analyses using the experimental design (July 2007–March 2008) suggested null effects on diarrhea and cough. However, children in the intervention villages had 32% reduced fever (relative risk, RR=0.68; 95% CI=0.62, 0.74). Use of Sprinkles was associated with about 30% reduction in diarrhea (RR=0.71; 95% CI=0.55, 0.93) and 18% reduction in fever (RR=0.82; 95% CI=0.72, 0.95).

Conclusion

Sale of Sprinkles may reduce fever and diarrhea incidence among children 6–59mo in western Kenya.

Grant Funding Source: NEVIN SCRIMSHAW INTERNATIONAL NUTRITION FOUNDATION
Lactobacillus rhamnosus GG inhibits neutrophil extracellular trap formation
Authors: Vong Linda , Lorentz Robert J. , Sousa Andrew J. , Glogauer Michael , Sherman Philip M.
Published: June 2021

Alterations in the gut microbiota, referred to as dysbiosis, are strongly associated with chronic inflammatory bowel diseases. Neutrophils are one of the earliest cells to respond to an inflammatory stimulus and, in the inflamed intestine, constitute a significant proportion of cells that are recruited. Whereas probiotics have been shown to exert beneficial effects on gut homeostasis, their effect on neutrophil extracellular trap (NET) formation remains undefined. Therefore, the aim of this study was to determine the effects of the probiotic Lactobacillus rhamnosus GG (LGG) on the dynamics of NET formation. Bone marrow‐derived neutrophils (BMDN) were isolated from C57BL6 mice and co‐incubated with LGG (1 h, 37°C, 5% CO2) prior to stimulation with phorbol 12‐myristate 13‐acetate (PMA, 100 nM), which is an inducer of NET formation. Measurements of extracellular DNA (Sytox Green, 5 nM) and immunofluorescence microscopy staining for DNA, elastase and histones demonstrated that LGG mediated a concentration‐dependent reduction in NET‐osis. Given the proximity of recruited neutrophils to the lumen of the inflamed gut and the resident intestinal microbiota, it is important to define how neutrophil‐microbe interactions affect the local microenvironment. This work was funded by CIHR (operating grant MOP‐89894).
Modulation of autophagy by bile acids in hepatocytes and liver
Authors: Manley Sharon Jessica , Guo Grace , Apte Udayan , Ding Wen-Xing
Published: June 2021

Autophagy is a lysosomal degradation process that regulates the homeostasis of organelles and proteins in cells. Mice with liver‐specific loss of autophagy develop spontaneous liver tumors possibly caused by p62 protein accumulation. Farnesoid X receptor (FXR) −/− mice exhibit higher hepatic bile acids (BAs) levels and develop spontaneous liver tumors. However, the mechanisms by which BAs promote liver tumors are not known. From the finding that FXR−/− mice have increased hepatic p62 proteins compared to wild type mice, which suggests that FXR−/− mice may have decreased liver autophagy, we hypothesized that BAs modulate autophagy and promote liver tumorigenesis. Using a series of autophagic flux assays, we found that hydrophobic and hydrophilic BAs differentially regulate autophagy, where hydrophobic BAs increase while hydrophilic BAs decrease autophagic flux in primary mouse hepatocytes. Interestingly, both hydrophobic and hydrophilic BAs increase p62 protein levels without significant changes to p62 mRNA expression. These results suggest that BAs‐induced accumulation of hepatic p62 proteins could be mediated by their differential modulation on autophagy, and current experiments are being conducted to determine these mechanisms. In conclusion, we found that hydrophobic and hydrophilic BAs differentially modulate autophagy, leading to the accumulation of p62 and LC3‐II proteins in hepatocytes.
Rational Selection of Antigens for Targeted Therapy of Liver Metastases
Authors: TURTOI Andrei , BLOMME Arnaud , CASTRONOVO Vincent
Published: June 2021

Patients suffering from liver metastases are diagnosed late and have a poor outcome. Targeted therapies are gaining a major stake in current and future treatment options. However, the malignant lesions are heterogeneous in nature offering niches for cancer cells causing treatment resistance and relapse. Therefore, a rational strategy is needed to select targetable antigens that would overcome this intra‐tumoral heterogeneity.

Here we generated a quantitative picture of the proteome heterogeneity in colorectal carcinoma liver metastases. We focus on membrane bound and extracellular proteins and show the differential distribution of novel targets and antigens against which the antibodies are already involved in clinical trials or treatment of liver metastases. Extensive clustering and validation experiments highlight novel markers that offer the potential to homogeneously cover the metastatic lesion and become better targets.

Two such antigens, LTBP2 and TGFBI are selected for functional analysis in colorectal carcinoma cells. In vitro and in vivo experiments show that both proteins are relevant for migration and proliferation capacities of cancer cells. Their depletion leads to significant inhibition of tumor growth, crystalizing them as bona fide targets for development of anti‐metastases therapies.

This work is funded by FP7 (ADAMANT) the European Union project, Univeristy of Liège & the ARC (IDEA) project, Belgium.
Transplantation of mesenchymal stem cells and fibroblasts in a bio‐compatible matrix corrects defective dermal remodeling
Authors: Yates-Binder Cecelia C , Rodrigues Melanie , Whaley Diana , Wells Alan
Published: June 2021

Curative molecular therapies are needed to correct matrix dysfunctions that results in scar formation upon healing. Mesenchymal stem cell transplantation has been reported to enhance healing however the mechanism for this response is unknown. The possibilities include the restorative paracrine effects of MSC synthesized soluble factors on dermal cells and matrix expression. This study examines the influence MSC's have on matrix remodeling in vivo. We recently developed a chronic wound scar model based on the absence of wound resolution through CXCR3 signaling axis; these mice develop sterile chronic inflammation and hypertrophic scars. Using fibroblasts encapsulated with MSC's in a biocompatible matrix, we determined that fibroblasts could overcome endogenous deficiencies and generate a more physiological dermal tissue. Xenotransplantation of CXCR3−/− mouse fibroblast and hMSC's into wounds of CXCR3−/− mouse scarring model revealed an increase in survival and reduced apoptosis, enhanced healing response, correction of matrix regeneration, and reduced inflammation during the first 30 days. Changes in matrix and soluble signals were also examined. These data suggest in situ matrix composition and mature matrix generation regulates scarring. This study provides insight into conceptual therapies of cell transplantation, by altering soluble factors, as viable for antifibrotic treatment.
Development of novel small molecules targeting β‐catenin driven hepatocellular carcinoma
Authors: Delgado Evan R , Mustata Gabriela , Monga Satdarshan Pal
Published: June 2021

Hepatocellular carcinoma (HCC) is associated with 90% mortality worldwide making it the 3rd leading cause of death by cancer. Aberrant β‐catenin activity occurs in 30% of HCC cases making it an attractive therapeutic target. Thus there is an urgent need to develop strategies to inhibit Wnt signaling. Here we report identification of several small molecules that inhibit β‐catenin activity utilizing an alternative drug discovery method. ZINC 8.0 database with approximately 3.5 million compounds was screened for morphologically similar drugs‐like subsets to an existing small molecule that inhibits β‐catenin/CBP interaction. The identified compounds were sorted and compounds showing similarity score equal or higher than 70% were retained. We identified 20 molecules after in silico ADME/toxicity filtering. Safety of top candidates was further tested on primary human hepatocytes and the compounds lacking toxicity at 200μM over 48 hours were tested further. Top 5 candidates were used at concentrations from 200 μM to 200 nM to assess effects on Wnt signaling in human hepatoma cells using TOPflash luciferase reporter assay and MTT viability assay. A significant decrease in TOPflash activity in HepG2 cells was observed as well as a decrease in cell viability and proliferation. Protein analysis showed a significant decrease in downstream β‐catenin targets such as cyclin‐D1 and Glutamine synthetase. One molecule showed efficacy at 200 nM and was screening was repeated resulting in 13 hits. Out of those, 7 molecules significantly reduced TOPflash at 100 μM. Thus novel drug discovery method, we have identified several molecules that specifically inhibit Wnt signaling in hepatoma cells and lead to their decreased survival and proliferation.
Development of liver fibrosis is critically regulated by phosphodiesterase 4 (PDE4) sub‐family
Authors: Gobejishvili Leila , Breitkopf Katja , Zhang Jingwen , Avila Diana , Dooley Steven S , Barve Shirish , McClain Craig J
Published: June 2021

The causal role of PDE4 sub‐families in the pathogenesis of lung disease has been extensively studied and PDE4 inhibitors have been demonstrated to be beneficial in the treatment of lung inflammation and fibrosis. However, mechanistic studies examining the role of PDEs in liver disease are lacking. Therefore, we examined potential pathogenic role of PDE4 in the development of liver inflammation and fibrosis using animal model of cholestatic liver injury. The obtained data demonstrate that initiation and progression of liver fibrosis is accompanied by a significant up‐regulation of PDE4A, B and D isoforms. Notably, treatment with the specific PDE4 inhibitor, rolipram, significantly decreased liver PDE4 activity, inflammatory cytokine expression, fibrosis and liver injury. Moreover, we show for the first time that primary hepatic stellate cells, which are major contributors to liver fibrosis, require PDE4B and D activity for their activation and production of extracellular matrix proteins. In summary, our results demonstrate that PDE4 isoforms are critically involved in the induction of expression of pro‐inflammatory and pro‐fibrogenic genes during development of liver inflammation and fibrosis. These data strongly suggest that inhibition of PDE4 activity represents a relevant therapeutic strategy for the treatment of hepatic inflammation, injury and fibrosis. The work was supported by NIH grants 1P01AA07103 and R01AA018869.
Vitamin D Standardization Program (VDSP)
Authors: Sempos Christopher T , Vesper Hubert W , Phinney Karen
Published: June 2021

The Vitamin D Standardization Program (VDSP) was begun to correct widespread variation in 25‐hydroxyvitamin D [25(OH)D] lab results that confounds international efforts to develop evidence‐based clinical & public health guidelines. Its objectives are to: (1) standardize measurement of total 25(OH)D in national health surveys world‐wide by linking them to the NIST reference measurement procedure (RMP); (2) study differences in 25(OH)D among those surveys; & (3) expand standardization services to include assay manufacturers & clinical & research labs. Measurands addressed in the VDSP are total 25(OH)D, 25(OH)D2, 25(OH)D3 & 3‐epi‐25(OH)D3. A formal laboratory standardization program will be implemented using procedures similar to those used in CDC Hormone Standardization (HoSt) Program. ODS provides overall coordination. A VDSP Coordinating Center has been established at CDC. NIST and Gent University (Dr. Thienpont), Belgium are the reference labs for the VDSP. Currently, VDSP is conducting an inter‐laboratory comparison study that includes labs from eight national surveys (Australia, Canada, Germany, Ireland, Mexico, South Korea, UK & US) as well as research labs & assay manufacturers to assess measurement variability in currently used assays. Data from this study will be used to create master equations to facilitate the conversion of already existing data from national health surveys to the NIST RMP.
Protein requirement in healthy pregnant women in early and late gestation determined by indicator amino acid oxidation method
Authors: Stephens Trina Victoria , Payne Magdalene , Ball Ronald , Pencharz Paul , Elango Rajavel
Published: June 2021

Protein (PRO) requirements in healthy pregnant women at 16–18 and 34–36 weeks gestation were determined using the indicator amino acid oxidation (IAAO) method. Nine, healthy women (age 25–34) each randomly received a different test PRO intake (range = 0.22–2.56 g/kg/day) during each study day (n=20). The diets were isocaloric and provided energy at 1.7 X resting energy expenditure (REE). PRO was given as a crystalline amino acid mixture based on egg protein composition, except phenylalanine and tyrosine, which were maintained constant across intakes. PRO requirements were determined by measuring the oxidation of L‐[1‐13C]‐phenylalanine to 13CO2 (F13CO2). Breath and urine samples were collected at baseline and isotopic steady state. Linear regression crossover analysis identified a breakpoint (requirement) at minimal F13CO2 in response to different PRO intakes.

Preliminary results indicate the mean PRO requirements in early and late gestation to be ~14% and 48%, respectively, above the estimated average requirement of 0.88 g/kg/day currently recommended by the Dietary Reference Intakes (DRI 2005) for macronutrients. Our results also indicate gestational differences in protein requirement, a consideration that has not been addressed by current DRI recommendations. This study is the first to directly estimate gestational protein requirement in a population composed solely of pregnant women, and suggests that current recommendations based on the nitrogen balance method may be underestimated.
Basal muscle protein synthesis is unaffected by sex in young and older adults
Authors: Markofski Melissa M , Timmerman Kyle L , Fujita Satoshi , Fry Christopher S , Glynn Erin L , Drummond Micah J , Dickinson Jared M , Reidy Paul T , Gundermann David M , Rasmussen Blake B , Volpi Elena
Published: June 2021

There are conflicting data in the literature regarding the effect of sex on muscle protein synthesis. However, the majority of studies had a small sample size and low power. Further, in some studies subjects had been hospitalized and on a standardized diet for up to 3 days prior to measuring protein synthesis. Others included obese subjects. Physical activity level, diet and adiposity may influence muscle protein synthesis.

METHODS

We pooled the fractional muscle protein synthesis rate (FSR) data collected by our lab over the past ten years (N=216) in non‐obese (BMI: 18.5–30.0 kg/m2), healthy young (18–40 yr; 74 males, 53 females) and older (60–87 yr; 57 males, 32 females) subjects using the same tracer (13C6‐ phenylalanine) and design (4‐hr primed‐continuous tracer infusion). All subjects maintained their normal activity level and diet until the night prior to the experiment.

RESULTS

There were no differences between all males (average±SD; 0.061±0.018%) and females (0.060±0.018%), young males (0.058±0.019%) and females (0.059±0.017%), or old males (0.064±0.015%) and females (0.061±0.022%). Overall, FSR was not significantly different between older (0.063±0.018%) and younger adults (0.058±0.018%), with no interactions between age and sex.

CONCLUSION

There are no sex differences in basal muscle protein synthesis between non‐obese, community‐dwelling, independent, young and older men and women.
Leucine balance, metabolic and satiety responses to a leucinerich meal in healthy elderly women
Authors: Snarr Chandra , Redmond Emily , Morais José A. , Wykes Linda J. , Chevalier Stéphanie
Published: June 2021

Many older adults experience a decline of appetite and may not meet their protein needs; a strategy to optimize protein retention is thus essential. Leucine has anabolic potential by stimulating protein synthesis via mRNA translation and insulin secretion. Postprandial responses to leucine‐rich versus isonitrogenous mixed meals were assessed in 9 healthy older women (70.1±2 y, BMI 23.7±2.7 kg/m2) by double‐blinded, randomized, crossover studies. Test meals provided 0.6 g protein and 16.0 kcal/kg LBM, once with added leucine (0.07 g/kg LBM), once without.

Results

Five‐hour whole‐body postprandial leucine oxidation response, measured by constant infusion of [1‐13C]leucine and enrichment of plasma 13C‐KIC and expired 13CO2 was further increased with meal+leucine. Yet, the resulting balance (intake‐oxidation) was greater (18.1±1.4 vs. without 11.3±1.3 mmol/5h, p<0.001) suggesting more protein retention. Serum insulin and C‐peptide AUC were greater post meal+leucine, indicating more secretion. Plasma glucose excursions and free fatty acids suppression did not differ between meals. Addition of leucine had no effect on meal‐induced thermogenesis, satiety and gut peptide responses (GLP‐1, PYY 3–36, ghrelin). This study shows the anabolic potential of a leucine‐rich mixed meal in older women, a dietary strategy that may be used to promote protein anabolism without affecting appetite. (Funded by CIHR)
Muscle breakdown determines Arginine (ARG) availability during hyperdynamic sepsis in the pig
Authors: Ten Have Gabriella A , Engelen Marielle P , Wolfe Robert R , Deutz Nicolaas E
Published: June 2021

RATIONALE

Arginine availability plays a key role in systemic ARG to nitric oxide (NO) conversion, but in sepsis it is unknown which of the organs contribute.

METHODS

Transorgan ARG kinetics was studied in 12 pigs with Pseudomonas aeroginosa induced hyperdynamic sepsis and 8 controls, using primed‐continuous Phenylalanine (13C6), ARG (guanido‐15N2) and Citrulline (CIT) (ureido‐13C‐5,5‐2H2) infusion. 18 h after start of the sepsis induction, whole body protein breakdown (WbPB), WbArg, WbCit, ARG de novo (WbdenovoArg) and nitric oxide (WbNO) production, and transorgan net balances (NB), disposal and production were measured. Enrichment and concentrations were analysed by LC‐MS. Data were reflected as mean (SE) (in nmol/kg bw/min); stats was done by RM‐ANOVA.

RESULTS

Sepsis increased WbPB, WbArg, WbNO (p<0.05), but did not change WbdenovoArg and WbCit. Only muscle ARG NB changed from net uptake to net release in the PM pigs (115(26) to −127(58); p=0.0046) and was caused by increased ARG production (326(157) to 762(107), p=0.028) and associated with increased muscle protein breakdown. No changes in gut, liver and lung ARG metabolism were observed.

CONCLUSION

ARG availability during sepsis is maintained only by stimulated muscle protein breakdown.
Impact of maternal prenatal metabolic abnormalities on metabolic hormones in human milk
Authors: Ley Sylvia H , Hanley Anthony J , Sermer Mathew , Zinman Bernard , O'Connor Deborah L
Published: June 2021

Metabolic hormones are present in human milk, but no studies have investigated the impact of maternal metabolic status during pregnancy on milk insulin and adiponectin concentrations. We aimed to investigate the association of prenatal metabolic status with milk insulin and adiponectin. Participants (n=170) underwent a 3‐h oral glucose tolerance test at 30 (95% CI 25, 33) weeks gestation and donated milk in the first week (early milk) and at 3 months postpartum (mature milk). Prenatal metabolic abnormalities including higher pregravid BMI (beta±SE 0.053±0.014, p=0.0003), in addition to gravid hyperglycemia (0.218±0.087, p=0.01), insulin resistance (0.255±0.047, p<0.0001), lower insulin sensitivity (−0.521±0.108, p<0.0001), and higher serum adiponectin (0.116±0.029, p<0.0001) were associated with higher insulin in mature milk with multiple adjustment. Prenatal metabolic measures were not associated with milk adiponectin, but obstetrical measures including nulliparity (0.171±0.058, p=0.004), longer duration of gestation (0.546±0.146, p=0.0002), and unscheduled C‐section (0.387±0.162, p=0.02) were associated with higher adiponectin in early milk with multiple adjustment. In conclusion, maternal prenatal metabolic abnormalities were associated with high insulin concentrations in mature milk, while only obstetrical parameters were associated adiponectin concentrations in early milk.

Grant Funding Source: CDA, CFDR, CIHR
Maternal body mass index modifies the impact of negative energy balance during lactation on infant weight and length gain in HIV‐exposed, uninfected Malawian females
Authors: Widen Elizabeth M , Bentley Margaret , Kayira Dumbani , Chasela Charles , Kacheche Zebrone , Tegha Gerald , Kamwendo Debbie , Jamieson Denise , Flax Valerie , van der Horst Charles , Adair Linda
Published: June 2021

Negative maternal energy balance during lactation may influence growth of exclusively breastfed infants through impaired quality or quantity of breastmilk. Malawian mother‐infant pairs (n=1,449) in the Breastfeeding, Antiretrovirals and Nutrition Study were randomized to receive a lipid‐based nutrient supplement (LNS), meeting nutritional needs of lactation, or no LNS and a maternal, infant or no antiretroviral (ARV) regimen. Linear regression models were used to relate negative maternal energy balance (weight loss 2–24 weeks) to infant weight and length gain from birth to 24 weeks, stratifying by gender and maternal BMI at 2 weeks (mean: 23.1 kg/m2), and controlling for birth measurement, parity, treatment arm and seasonality. Length (β: −3 cm, p=0.01) and weight gain (β: −1.3 kg, p=0.001) were lower in girls whose mothers had lower BMI at 2 weeks postpartum coupled with negative energy balance, compared to women in positive energy balance. Treatment arm was not associated with infant length or weight gain (p>0.13). Though effects were only observed in girls, suggesting possible gender differences in suckling and feeding behavior, these findings indicate that maternal negative energy balance with low energy reserves represents a risk factor for poor infant growth outcomes, possibly through reduced milk energy output.

Grant Funding Source : CDC(U48 DP000059 01); Bill & Melinda Gates Foundation(OPP53107); CPC(NICHD 5 R24 HD050924)
Development of resting cardiovascular activity during the first 2 years of life differs in breastfed and formula‐fed boys and girls
Authors: Pivik R T , Andres A , Badger T M
Published: June 2021

To investigate whether early infant diet influences cardiovascular development we recorded resting heart‐rate (HR) at 3, 6, 9, 12, and 24 mo in awake healthy children who were breastfed (BF) or fed milk formula (MF) or soy formula (SF) during infancy (n = 83–146/group/age). HR, and indices of autonomic nervous system (ANS) sympathetic [low frequency power (Plf: .04–.15 Hz)] and parasympathetic activity [high frequency power (Phf: .15–.5 Hz)] were determined from power spectral analyses of digitized, artifact‐free recordings and analyzed using ANOVAs with post‐hoc t‐tests. HR decreased and Plf and Phf increased for all groups across the study period. Few within‐group gender differences were found [6 mo Plf: MF girls > boys, p < .05; 12 and 24 mo HR: SF girls > boys, both p < .05]. There were three important diet‐related findings: 1) levels of Plf and Phf were higher in BF than formulafed (FF) groups after 6 mo (p < .001); 2) SF and MF infants did not differ on HR measures; 3) and, group differences were modulated by gender [greater Plf and Phf in FF than BF girls from 9–24 mo (p < .02) and in FF than BF boys at 12 and 24 mo (p < .01)]. Baseline levels of ANS activity influence interactions between the central nervous system and ANS that regulate attention, emotion, cognition and behavior. The present findings suggest early infant diet may contribute to individual differences in these functions.

Supported by USDA ARS CRIS 6251‐51000‐006‐03S
A Morphometric Study of Phylogenetic and Ecologic Patterns in Procyonid (Mammalia: Carnivora) Endocasts
Authors: Ahrens Heather Elizabeth , DeLeon Valerie B
Published: June 2021

Brain size and shape have been linked to numerous evolutionary trends, with patterns of encephalization and variation well‐studied in Carnivora. Endocasts provide a proxy for brain morphology and access to potential suites of characters, but rarely are endocast characters used in cladistic analyses. We used three‐dimensional geometric morphometrics to examine phylogenetic and ecologic patterns in the endocasts of procyonids. Endocasts of seven ingroup taxa and four outgroup taxa were digitally rendered and 21 surface landmarks were collected. We tested correspondence of ‘brain shape’ with competing morphological and molecular phylogenies of Procyonidae. In analyses incorporating all taxa there was phylogenetic signal in brain shape for both topologies, but in analyses of ingroup taxa there was phylogenetic signal in the morphological topology only. Differences in brain shape suggest that procyonids emphasized areas associated with motor control, spatial perception, and balance relative to the basal musteloid, perhaps associated with increased arboreality. In Procyonidae, areas of the brain associated with arboreal behavior are also expanded in some taxa, but greater variation of brain shape occurs.

Grant Funding Source: none
Exceeding expectations: Examining the systemic effects of high body mass in relation to mechanical loading
Authors: Reeves Nicole M.
Published: June 2021

In studies of the effects of body mass on the human skeleton, an emphasis is often placed on the principle of bone functional adaptation in limb bones, namely in response to mechanical loading. However, it is possible that high body weight has systemic effects on bone throughout the skeleton, both due to increased loads and physiological effects, such as hormonal levels. Studies have correlated severe obesity with changes in joint morphology and some cross‐sectional properties of lower limb bones. This study adds further analyses by examining effects on the upper limb, which likely experiences increased mechanical loads in obese individuals, as well as the cranial vault, which experiences nominal loading. All data were collected from high‐resolution tomographic scans of modern individuals from the William M. Bass Donated Collection at the University of Tennessee, Knoxville. Cross‐sectional properties were taken from multiple diaphyseal locations on the humerus, radius, femur and tibia on human remains from people known to be within normal clinical weight ranges and people documented as overweight. Results suggest a systemic effect of obesity on the skeleton, although the magnitude of the effects is greatest in bones supporting body mass. Cranial vault thickness produces more variable results. Further analysis will consider variation in joint size and shape of the shoulder, hip and knee.

Grant Funding Source : AAA
Adaptations of the musculoskeletal system in California sea lions to the semi‐aquatic lifestyle
Authors: Bragulla Hermann Heinrich , Schwartz Jillian R
Published: June 2021

The musculoskeletal system of California sea lions shows peculiar morphological adaptations, which enables rapid turns when swimming, but also allows weight bearing when walking on land. Statements in the literature regarding how sea lions are able to walk on land are contradictory. The range of motion of the pelvic limb is limited by the fact that thigh and crus are enclosed in the body wall. The objective of this study is to analyze the musculoskeletal system of the vertebral column, pelvis, and pelvic limbs in sea lions. California sea lions were obtained from “The Marine Mammal Center” in Sausalito, California. Prior to dissection, series of computed topographic images (CTs) of the pelvis and pelvic limbs in the flexed (walking), intermediate, and extended (swimming) position were acquired. Virtual 3D‐reconstructions of the skeleton were analyzed to visualize the actual range of motion of the vertebral column, pelvis, and pelvic limbs. Videos of sea lions were analyzed frame by frame to understand the range of pelvic limb motion in the transition from swimming to walking. For walking and weight bearing on the pelvic limbs, the hip joint is slightly flexed, the femur is adducted, and the angle of the stifle remains relatively unchanged, while tibia and fibula, as well as the tarsus are rotated inwards. The tarsus enables a unique rotation of the foot, which is not seen in terrestrial, but is in arboreal mammals.
A novel skeletal disorder defines an intracellular role for FGFR2 during development
Authors: Merrill Amy Elizabeth , Sarukhanov Anna , Krejci Pavel , Idoni Brian , Lachman Ralph , Wilcox William , Krakow Deborah
Published: June 2021

Fibroblast growth factor (FGF) signaling critically regulates bone formation during embryonic development. This is exemplified in craniosynostosis syndromes resulting from activating mutations in Fibroblast Growth Factor Receptor 2 (FGFR2). While it is known that FGFR2 coordinates osteoprogenitor cell proliferation and differentiation, it remains unclear how the receptor couples these distinct cellular processes. Through delineation of a novel lethal skeletal disorder with FGFR2 mutations, we have unexpectedly revealed that the receptor directly promotes ribosomal biogenesis, a process essential for proliferation. The dominant FGFR2 mutations we identified selectively reduce plasma membrane levels of FGFR2 and markedly diminish receptor responsiveness to extracellular FGF2. Interestingly, we found that mutant FGFR2 enhances responsiveness to intracellular FGF2 and enriches nucleolar association with FGF2 and the ribosomal RNA transcription factor UBF1. This activity consequently elevates rRNA expression and blocks nucleolar occupancy of the osteoblast regulator RUNX2. As RUNX2 reduces rRNA transcription during osteoblast commitment, these data explain the skeletal findings of the new disorder. By executing activities in the cell membrane and nucleolus, FGFR2 integrates intracellular and extracellular FGF signals to balance osteoprogenitor cell proliferation and differentiation.
Protection against molecular aging and degeneration
Authors: Clark John I
Published: June 2021

Image formation in vertebrate vision depends on transmission and focusing of light by the biological lens. The biophysical properties of visible light constrain the differentiation of cells in the embryonic surface ectoderm during the formation of symmetric, concentric, transparent layers of variable index of refraction. In the absence of turnover, lens cells and their molecular constituents are retained for the lifetime of the individual and a robust biological mechanism is necessary to protect against protein aggregation resulting from the cellular stress of aging.

AlphaB crystallin is part of a selective rather than specific mechanism for recognition of protein instability. Patterns of interactive sequences recognize surface instabilities responsible for the self assembly of normal intracellular filaments or abnormal fibrils and aggregates. There is much to learn about the cooperative nature of the interactive surface domains both in the normal assembly of structural proteins and in the self assembly of abnormal fibrils and aggregates. Supported by EY04542 from the N.E.I.
Using genomic technologies to investigate transcriptional regulation in normal and cancer cells
Authors: Farnham Peggy J
Published: June 2021

As part of the ENCODE Consortium, we have studied TCF7L2, a transcription factor linked to a variety of human diseases such as Type 2 diabetes and cancer. One mechanism by which TCF7L2 could impact such diverse diseases is by binding to different genomic regions in different tissues. To test this hypothesis, we performed ChIP‐seq for TCF7L2 in 6 human cell lines, identifying 98,361 binding sites, with only 1,578 sites common to all 6 cells. To investigate cell type‐specific binding, we searched for motifs enriched in the cell type‐specific sites, finding a motif for HNF4alpha in the TCF7L2 sites from HepG2 cells and the GATA3 motif in TCF7L2 sites from MCF7 cells. ChIP‐seq analysis revealed that TCF7L2 co‐localizes with HNF4alpha in HepG2 cells and with GATA3 in MCF7 cells. As a further indication of the importance of TCF7L2, we found that it binds within 10 kb of 50% of all single nucleotide polymorphisms in the GWAS Catalog. Our studies reveal important insight into TCF7L2‐mediated gene regulation and suggest that TCF7L2 plays critical roles in specifying cellular phenotypes. This study was supported by funds from Public Health Service CA45250 and 1U54HG004558.
BRCA1 and CtIP are required for processing uncapped telomeres
Authors: Tarsounas Madalena
Published: June 2021

BRCA1 tumour suppressor is a large phosphoprotein with key roles in the surveillance of genome integrity. As an E3 ubiquitylation enzyme, BRCA1 transduces DNA damage signals to the effector checkpoint kinases to arrest cell cycle progression and engage the homologous recombination pathway of DNA repair. A target of BRCA1‐dependent ubiquitylation is CtIP, which stimulates the nucleolytic activity of the MRE11‐RAD50‐NBS1 complex in vitro. BRCA1 is thought to promote resection at DNA double‐strand breaks by activating CtIP, a process counteracted by 53BP1. Here, we demonstrate that both BRCA1 and CtIP are required for the processing of DNA ends generated by telomere uncapping. Loss of the telomeric factor TRF2 in mouse embryonic fibroblasts causes disassembly of end protective structures, leading to chromosome fusions. Conditional Brca1 deletion or CtIP inhibition partially abrogates telomeric fusions in cell cycle‐dependent manner. BRCA1 and CtIP activities are specifically required during the post‐replicative processing of uncapped telomeres. Thus, in addition to non‐homologous end‐joining, BRCA1‐mediated repair facilitates processing of dysfunctional telomeres leading to genome instability.
Molecular insights into tether function at the late endosome and vacuole
Authors: Ungermann Christian
Published: June 2021

Membrane fusion within the eukaryotic endomembrane system depends on the initial recognition of Rab GTPase on transport vesicles by multisubunit tethering complexes (MTC) and subsequent coupling to SNARE‐mediated fusion. The conserved vacuolar/lysosomal HOPS tethering complex combines both activities. Here, I will present insights into HOPS targeting to endosomal membranes, its regulation by phosphorylation, and its overall structure. Our data reveal that HOPS has a flexible elongated structure. I will show the localization of the individual subunits and the Rab binding site. Based on these findings, I will present a model how HOPS can tether membranes and support membrane fusion at late endosomes and vacuoles.
Apoprotein B: quality control early and late in the secretory pathway for this atherogenic protein
Authors: Fisher Edward A , Ginsberg Henry N , Ron David , Williams Kevin J , Brodsky Jeffrey L
Published: June 2021

Apoprotein B (apoB100) is required for the formation of VLDL, the precursor of LDL, the plasma level of which is a strong risk factor for atherosclerosis and coronary artery disease (CAD). It is an exceptionally large (>4000 aa) and hydrophobic protein. As the nascent polypeptide is translocating across the ER, if lipids are not transferred to stabilize its conformation, we and others have shown that like other abnormally folded proteins in the ER, its quality control is governed by ER‐associated ub‐proteasome degradation (ERAD). Using a combination of systems from yeast to genetically modified mice, we and our collaborators have shown apoB100 ERAD to involve chaperone and chaperone‐like molecules, such as Hsp70, 90, 110, and p58. The full assembly of VLDL does not occur until the Golgi. We have discovered that nascent particles (“pre‐VLDL”) that do not complete this maturation process (e.g., under the influence of certain dietary fatty acids), are subject to a late stage quality control process mediated by autophagy. In summary, the production of apoB100‐lipoproteins, whose plasma levels correlate with the risk of CAD, is determined in part by quality control processes that recognize features of the protein specific to its stage of assembly and that are separated between the ER and Golgi compartments.
EWS/Fli1‐regulated microRNAs in Ewing Sarcoma pathogenesis
Authors: Jedlicka Paul , Parrish Janet , McKinsey Erin , Dylla Layne , Niemeyer Brian , Moore Colin
Published: June 2021

Ewing Sarcoma is an aggressive malignancy with poor long‐term outcome. In an effort to identify more effective therapeutic approaches for Ewing Sarcoma, our laboratory has been studying the biology of microRNAs, novel regulators of gene expression with therapeutic potential. As a first approach, we have focused on microRNAs downstream of the EWS/Fli1 fusion oncoprotein, the dominant oncogenic driver in Ewing Sarcoma. Initial studies have identified growth suppressive microRNAs that target the IGF signaling pathway, an important tumor‐promoting axis in Ewing Sarcoma. Ongoing studies seek to understand: 1) how these and other microRNAs contribute to Ewing Sarcoma pathogenesis; 2) how the expression of microRNAs is regulated by the EWS/Fli1 oncoprotein; and 3) how these findings could be harnessed for novel therapies for this aggressive disease.

This work has been supported by the Boettcher Foundation's Webb‐Waring Biomedical Research Program, Alex's Lemonade Stand Foundation for Childhood Cancer, Ruth L. Kirschstein National Research Service Award Institutional Training Grant, and the University of Colorado Cancer Center and School of Medicine.
Functional Genomics Of Endothelial Cells Treated With Chemopreventive Anti‐Angiogenic Drugs
Authors: Sogno Ilaria , Cantelmo Anna Rita , Bartolini Desirée , Noonan Douglas M. , Albini Adriana
Published: June 2021

Many chemoprevention agents and substances (including dietary habits) associated with cancer prevention appear to target angiogenesis, strengthening the concept that “angioprevention” may represent a feasible and efficacious approach to preventing cancer. However the pathways modulated in the endothelial cells by these potential antiangiogenic chemopreventive drugs (CDDO triterpenoids, 4HPR, xanthohumols, ECGC, curcumin, hyperforin) are still unknown. Microarray‐based studies have been carried out on endothelial cells treated with selected chemopreventive compounds with anti‐angiogenic effect. The NFêB, TNF‐á, TGF‐â, Akt and MAPK pathways emerge as central targets shared by many of the drugs tested. Effects have been compared to those of endogenous angiogenesis inhibitors (as angiostain, endostatin), cytokines (INFs). The knowledge of the targets activated or repressed in treated endothelial cells, as provided by functional genomics studies, can help to identify new molecular targets and potentially synergistic drug combinations to be applied in clinical practice.
Integration of technology and science education: A collaborative workshop for instructors
Authors: Hurtt Barbekka C , Kipp Brian H. , Dutta Kaushik
Published: June 2021

This workshop focuses on the benefits and challenges of incorporating digital education technology into 21st century undergraduate science education. Students and faculty alike are adapting to a new, more technologically driven paradigm of pedagogy in the ‘classroom.’ In The College of 2020: Students (2009), Van Der Werf and Sabatier predict this shift will continue in a variety of manners: greater student access of online courses, improved access via portable computing devices, greater connectivity and creativity from colleges, personalization of education, exploring concepts through games and simulations, among other pedagogical changes. Scientific organizations, publishing companies, and technology companies are driving many of these technological innovations. Research is starting to emerge on the pedagogical effectiveness of digital education resources, although no clear, definitive relationship has been identified. The goals of this workshop are to discuss: 1) how digital education resources can be leveraged to complement and enhance traditional science education pedagogical approaches, and 2) preliminary results of studies identifying the resources anatomy and physiology students find most effective for studying and learning. Digital content from McGraw‐Hill Higher Education as utilized by two faculty members in their A&P courses will be discussed as part of the preliminary research.
Using open source technology to enhance nursing students’ understanding of physiology
Authors: Lafuze Joan Esterline , Runshe Debra Dunlap
Published: June 2021

Nursing faculty on our campus often report they need to re‐teach what has been taught in physiology and courses that serve as pre‐or co‐requisite to the study of physiology. Students may have learned information well enough to take tests, but they have not integrated the material well beyond learning facts nor transported it to a “place” where they can share it. Using a number of the open course, Sakai, tools engages nursing students in their own learning experience. By adding the Sakai ePortfolio tools, students have an opportunity to reflect about their learning in preparation for clinical experiences. The ePortfolio tools allow opportunities for students to reflect and synthesize this information and express it in a way that prepares them for their clinical experiences. We will discuss simple ways for others to introduce students to “folio thinking” with the Sakai ePortfolio tools in this presentation. Several other Sakai tools that are being used will also be discussed such as: the Assignments, Modules, Resources, Chat, and the Samigo Test and Survey. These tools provide students access to course lectures, videos, publisher materials, and virtual labs. Students have responded positively to this teaching strategy. Student submissions reflect deeper understanding of the concepts through connections to their clinical experiences. The research question: The collaboration between basic anatomy and physiology instruction and a school of nursing faculty effect student learning. We will measure student and faculty satisfaction, changes, in retention and progress in the nursing program. Long term goals include: to follow nursing graduates in later employment and assess their perception of how their knowledge of the basic sciences has assisted them in their positions. The source of research support is local funding.
The relationship between oxidative SOD1 and Abeta production and aggregation
Authors: Li Chen , Chen Xueping , Li Xin-min , Kong Jiming
Published: June 2021

To determine the relationship between SOD1 oxidation and amyloid beta production and aggregation, we used ELISA to analysis soluble and insoluble Aβ40 and Aβ42 in the cortex and spinal cord and used immunofluorescence to test the co‐localization of the oxidative SOD1 aggregation and Aβ aggregation. We found that the insoluble Aβ40 in cortex was significant higher in 6 month old G37R mice than in non‐transgenic mice. The soluble and insoluble Aβ42 in cortex and spinal cord were significantly higher in 6 month old G37R mice and G93A mice than in non‐transgenic mice. The mutant SOD1 could influence the production of the soluble or insoluble Aβ40 and Aβ42 in the brain and in the spinal cord. The oxidative SOD1 aggregations and Aβ40 were co‐localized in the G93A and G37R transgenic mice, but not non‐transgenic mice. Meanwhile, the oxidative SOD1 aggregations localized in the senile plaques in the APP/PS1 mice. We speculated that mutant SOD1 was easy to be oxidated, and the oxidative SOD1 could interact with APP. SOD1 oxidation may be the earliest event that triggers SOD1 aggregation and induces Aβ aggregation. We will cross APP transgenic mice with G93A and G37R mice to further prove our conclusions.
Effects of heroin dependence on the expression of L‐ENK and SS in the VTA and NAc regions of rat brain
Authors: Li Yi-xin , Liang Wen-mei
Published: June 2021

To investigate the effect of the heroin addicts on ratˈs L‐ENK‐ ¡¢SS‐immunoreactive cells in the ventral tegmental area(VTA) and nucleus accumbens(NAc) regions, and exploring the effect of heroin dependence on learning and memory mechanisms. Adulthood 55 male SD rats were randomly divided into normal control group(NCG), saline control group(SCG) and the heroin‐dependent group(HDG). Heroin‐dependent rat models were set up by the way of subcutaneous injection of heroin and brain tissues are excised on the 10th, 17th, 24th, 31th, and 38th days after models are set up. Immunohistochemical staining was performed to test the expression of L‐ENK and SS in the VTA and NAc regions. The expression of L‐ENK and SS was observed as brown yellow particle in the cytoplasm of neurons in the VTA and NAc regions. L‐ENK‐immunoreactive cells and SS‐immunoreactive cells expressed less L‐ENK and SS in the VTA and NAc regions from HDG rats than that from SCG and NCG rats at different time points. Immunohistochemical analysis showed higher grey value was found in HDG rats than that in controls (p<0.05). The results suggest that the secretion of L‐ENK and SS is inhibited during heroin dependence, and heroin dependence may cause damage to learning and memory function.