bluuebunny/crossref_metadata_2025 huggingface dataset:
Risk factors for folate deficiency differ from those for vitamin B12 deficiency in Cameroonian women and children (119.7)
Authors: Shahab‐Ferdows Setareh , Engle‐Stone Reina , Hampel Daniela , Ndjebayi Alex , Nankap Martin , Brown Kenneth , Allen Lindsay
Published: June 2021

To assess nutritional status and risk factors for deficiency prior to a national fortification program, a nationally representative survey was conducted in 3 ecologic zones of Cameroon (South, North, and Cities). 24h dietary recall data were collected from 10 households (HH) in each of 30 randomly selected clusters/zone.. In a subset of women of reproductive age and their 12‐59 mo old children (n= 396 pairs), plasma B12 & folate were analyzed by radioassay. Based on plasma values, 17% and 29% of women and 8% and 30% of children were folate (<10nm/L) and B12 (<221pm/L) deficient respectively. Folate deficiency did not differ among zones, but B12 deficiency was higher in the North/ vs. Cities. Folate and B12 dietary intake patterns were similar for women and children. Risk factors for folate and B12 deficiency differed: plasma folate was higher in the South; B12 was higher in the Cities. Folate intake was higher in the North vs. South/Cities and among those with low SES vs. high SES. B12 intake was highest in Cities and among those with higher SES, and lower in the rural vs. urban areas. Breast milk B12 was lowest in the North vs. Cities and South and correlated with B12 intake. Overall inadequate folate status is a problem in the Cities while low B12 intake and status are more common in poor and rural zones. The differing risk factors for folate and B12 deficiency have implications for folic acid and B12 fortification.

Grant Funding Source: Michael and Susan Dell Foundation, UNICEF and Bill & Melinda Gates Foundation
Plasma amino acid concentrations following a beef‐rich lunch meal and the effects on appetite control and satiety in healthy adults (120.1)
Authors: Ortinau Laura , Lasley Tyler , Douglas Steve , Leidy Heather
Published: June 2021

Background: The consumption of dietary protein leads to improved appetite and satiety. However, it is unclear as to the actual nutrient signal that elicits these responses. Purpose: To identify the amino acids (AAs) that serve as nutrient signals for appetite and satiety following a beef‐rich lunch meal. Methods: Twenty‐three adults (age 23 ± 1 y; BMI: 23.4 ± 0.6 kg/m2) consumed a 400 kcal beef‐rich lunch meal containing 34 g of protein. Questionnaires and blood samples were collected at baseline and every 60 min thereafter for 180 min. Post‐lunch outcomes included perceived hunger and fullness and plasma ghrelin, GLP‐1, PYY, and AA concentrations. Results: Multivariate regression analyses revealed that post‐lunch plasma AAs concentrations accounted for only 10% of the variability in hunger (p<0.01) and 16% of the variability in fullness (p<0.001). However, post‐lunch plasma AAs concentrations accounted for 57% of the variability in PYY (p<0.001), 69% of the variability in GLP‐1 (p<0.001), and 52% of the variability in ghrelin concentrations (p<0.001). The essential AAs eliciting these effects were threonine, lysine, phenylalanine, and valine. Conclusion: Although plasma AA concentrations following a beef‐rich lunch meal elicit very little effect on perceived sensations, they do exert a substantial effect on the gastro‐intestinal hormones associated with appetite control and satiety in healthy adults.

Grant Funding Source: Supported by: The Beef Checkoff
Diets diet rich in stearidonic acid negatively impact visceral adipose mass (121.1)
Authors: Kraft Jana , Bay Pamela , Rowlandson Shae , Peshavaria Mina , Jetton Thomas
Published: June 2021

Dietary marine‐based n‐3 fatty acids have been implicated to have beneficial effects on insulin resistance and type 2 diabetes. The objective of the present study was to determine if consumption of stearidonic acid, from a terrestrial plant‐based (Echium oil) n‐3 fatty acid, may impact diet‐induced obesity and glucose homeostasis in mice. B6 mice were fed a low‐fat diet (LF, 10% kcal total fat) or high‐fat diet (HF,60% kcal total fat) containing 20% of total fat content of either fish oil (FO), Echium oil (EO), or no supplement (Control, C) for 12 weeks. Body mass and daily weight gain were ~50% greater in HF than in LF mice (P < 0.001), but no differences were observed within the groups. Although food intake was not different between the groups, energy intake was greater (+38%) in HF than LF mice. EO‐fed mice exhibited a lower fat pad/body weight ratio when compared to control mice. Pancreas weight and pancreas/body weight ratio was lower in the HF‐EO mice when compared to HF‐FO (P < 0.05). As expected, plasma glucose and insulin were increased in HF mice when compared to LF (P < 0.05); however, no differences within the LF or HF groups were observed. Glucose and insulin tolerance tests showed impaired glucose tolerance and insulin resistance, respectively, in HF mice when compared to LF mice, but no oil‐specific effects. Hence, diets rich in stearidonic acid appear to impact tissue energy distribution by decreasing visceral adipose mass.
Supplementation of polyphenol‐rich blackcurrant extract exerted hypolipidemic and anti‐inflammatory effects in diet‐induced obese mice (121.7)
Authors: Benn Tyler , Kim Bohkyung , Park Young‐Ki , Ku Chai Siah , Yang Yue , Pham Tho , Wegner Casey , Faruggia Callie , Harness Ellen , Lee Ji‐Young
Published: June 2021

Obesity is closely tied to hyperlipidemia, chronic inflammation, and insulin resistance. To gain insight into the effect of polyphenol‐rich blackcurrant extract (BCE) on obesity‐associated abnormalities, we fed 24 male C57BL/6J mice a high fat/high cholesterol diet (15% fat, 0.2% cholesterol by wt) supplemented with 0.1% BCE for 12 wk. Plasma total cholesterol and glucose were significantly reduced with marked increases in hepatic expression of sterol regulatory element binding protein 2 and low‐density lipoprotein receptor in BCE‐fed mice. Liver histology demonstrated less visible steatosis in BCE group. The body weight % of epididymal fat pads was significantly less in BCE‐fed mice compared to control. Furthermore, adipocyte size was smaller and the number of crown‐like structures, a marker of macrophage infiltration, was fewer in conjunction with decreased mRNA of the macrophage markers F4/80 and CD68 in epididymal adipose of BCE‐fed mice. Ex vivo analysis of splenocytes isolated from control and BCE‐fed mice showed that upon lipopolysaccharide stimulation, the expression of tumor necrosis factor α and interleukin 1 β were significantly reduced in the BCE group. In conclusion, BCE supplementation may be beneficial to prevent metabolic disturbance and chronic inflammation in obesity.
Iron absorption in vegetarian children is decreased at higher calcium intakes (122.7)
Authors: Abrams Steven , Hawthorne Keli , Chen Zhensheng
Published: June 2021

Vegetarian children may be at risk for poor iron (Fe) status due to either low intake or decreased bioavailability of dietary Fe. Using the stable isotope 57Fe, we measured Fe absorption in a group of vegetarian children 4 to 10 years of age (Mean 7.1 ± 2.1 y), with dietary Fe intakes averaging 13 ± 4.3 mg/d. Mean Fe absorption was 9.6 ± 6.6 % (SD) and mean serum ferritin was 27.4 ± 13.7 mg/mL. We found significant predictors of Fe absorption (n=29) to be age (t = ‐2.5, P < 0.01), transferrin saturation (TSat) (t = ‐3.8, P < 0.001), energy intake (t=3.0, P < 0.01), and calcium (Ca) intake (t = ‐3.9, P < 0.001). Dietary Fe, gender, body weight, height, and serum ferritin were not significantly related to Fe absorption efficiency (% absorption). Overall, 15 of 29 children had a TSat < 20% (8 were <15%) and 10 had a serum ferritin < 20 mg/mL. Those (n = 16) with dietary Ca intakes above 700 mg/day had a 15% Fe absorption whereas those with a Ca intake below that (n = 13) had a 5% Fe absorption (p < 0.0001) after adjustment for age, TSat, and energy intake. These data indicate that vegetarian children on self‐selected diets are at risk for poor Fe status that may be mediated in part by higher Ca intakes including those within the range of dietary recommendations. Although overall Fe absorption was adequate to meet growth requirements, the need for calcium to meet bone health requirements suggests that vegetarian children with only non‐heme Fe intake may require higher dietary intakes of Fe.
The aerodynamics of feather asymmetry and implications for paravian flight (13.4)
Authors: Hall Justin
Published: June 2021

The presence of asymmetric feathers has been regularly used as an indicator of aerodynamic function and flight capacity in fossil birds and non‐avian dinosaurs. However, most of the flight feathers in living flying birds are not highly asymmetric. Only the most lateral primaries are typically highly asymmetric, and secondaries are typically symmetrical. The effects of feather asymmetry are more complex than simple presence or absence metrics indicate. Complex twisting occurs in the lateral primaries of living birds, which delays stall, improves aeroelastic stability, and promotes dynamic maneuvers and rigorous flapping flight at low speeds. Modeling feathers as simple, straight, flat plates suggests that they will only twist in the stabilizing “wash out” manner if vane depth ratios exceed 4:1. Because the center of lift on a flat plate sits near 1/4 chord, feathers with vane depth ratios of 4:1 or less are loaded primarily in bending. I demonstrate that the lateral primaries in most species are asymmetric enough to twist in wash out, as anticipated under the straight plate model. However, a substantial number of species do not meet expectations, which demonstrates that some combination of feather overlap, curvature, and orientation works to help stabilize these feathers under aerodynamic load. These patterns can be applied to fossil taxa to draw more precise conclusions about wing, hindwing, and tail functions.
Formula use in a breastfeeding culture: changing perceptions and patterns of young infant feeding in Vietnam (131.6)
Authors: Henry Megan , Christian Parul , Kim Sunny , Pham Van , Nguyen Nam , Nguyen Phuong , Rawat Rahul , Menon Purnima
Published: June 2021

Previous research identified an increase in formula use as a barrier to exclusive breastfeeding (EBF) in Vietnam. In the context of a process evaluation (PE) of the Alive & Thrive program to promote EBF, 121 in‐depth interviews with rural mothers, fathers, and grandmothers of infants 0‐12 mo formed the basis of a case study analysis (n=48 families). While 79% of mothers had heard about EBF and breastfeeding was universal, EBF prevalence was only 2%. Two‐thirds of infants were formula fed (FF) before 6 months (40% at birth, 12% at 1‐3 mo, and 15% at 4‐6 mo). Stopping and restarting FF was common. Among early starters, families reported a desire to support the mother, “what I could do for her was feed our baby with formula,” often sua non (bovine colostrum formula) sold in the hospital. All 6 cesarean sections FF at birth. Mothers were reluctant to FF but felt they had “no choice” when infants were “crying because of hunger”. For families introducing FF at 1‐3 mo, perception of insufficient milk (PIM) drove use. For the category of 4‐6 mo, introduction of complementary food (CF) was also early due to pressure from grandmothers, or returning to work, or under the perception that infants need to be prepared to accept CF. High awareness of formula advertisements, conflicting advice from health professionals, and a universal perception that insufficient maternal diet led to poor breastmilk supply also influenced FF despite the high cost. Families valued and practiced BF, but require targeted support for BF initiation and PIM, as well as appropriate and timely CF.

Grant Funding Source: Alive & Thrive, funded by the Bill & Melinda Gates Foundation and managed by FHI360
Development and piloting of a multi‐level obesity prevention program for low income African American children: The B’More Healthy Communities for Kids study (132.4)
Authors: Gittelsohn Joel , Anderson Steeves Elizabeth , Mui Yeeli , Kharmats Anna , Dennis Donna , Henry JaWanna , Hopkins Laura , Wyse Ransom
Published: June 2021

Multi‐component interventions are needed in low income urban settings to address the complex environmental, economic, and social issues driving obesity. Low income neighborhoods in Baltimore can be described as “food swamps,” with few supermarkets and many small food outlets stocking unhealthy foods. After a decade of focused work in specific food source types (e.g. corner stores) in Baltimore, we are developing and implementing a multi‐level intervention to prevent childhood obesity. Substantial formative research has been conducted to develop the B’More Healthy Communities for Kids (BHCK) program, including in‐depth interviews (n=24) with children, parents and store owners, focus groups (n=4), and direct observations (n=7). We piloted structural incentives for small sources, a farm to store component, and a peer mentored cooking program. The BHCK program will be implemented at the policy, wholesaler, corner store, carryout, recreation center, family and individual levels. The program will use a four‐fold strategy to improve healthy eating behavior ‐ including environmental interventions to increase access to healthy foods, behavioral nudging to make choosing healthier options easier, interactive nutrition education opportunities to increase consumer demand, and policies (informed by systems models) to increase long‐term sustainability. Detailed process and impact evaluations will take place at each intervention level.

Grant Funding Source: NIH U54
Folic acid in a high multivitamin diet during pregnancy increases post‐weaning weight gain and PPAR gene expression in adipose and liver of Wistar rat dams (135.2)
Authors: Pannia Emanuela , Cho Clara , Sanchez‐Hernandez Diana , Huot Pedro , Poon Abraham , Kubant Ruslan , Anderson G
Published: June 2021

High multivitamin (10‐fold, HV) or folic acid (Fol) intake alone during pregnancy leads to an obesogenic phenotype in the offspring but its effects on the later post‐weaning (PW) weight gain of the dams has not been reported. This study investigated whether Fol alone as a component of a HV gestational diet affects PW weight gain and associates with expression of peroxisome‐proliferator activated receptor isoforms (PPAR‐α/β/γ) involved in lipid metabolism and adipocyte differentiation. Wistar rat dams were fed an AIN‐93G diet during pregnancy containing either: 1) Recommended vitamin (RV); 2) HV; or 3) HV with recommended Fol (HVNF) content (n=15/group). Dams were fed a RV diet during lactation and then switched to a high fat (RV with 60% fat) diet for 16 weeks. Dams fed the HV diet during pregnancy gained 30% more weight over time from weaning (p<0.05) and had increased PPAR‐γ (3.8‐fold, p<0.05) and PPAR‐α (1.2‐fold, p<0.05) mRNA expression in visceral adipose and liver respectively compared to RV dams. Reducing Fol to recommended quantities modulated these effects as HVNF dams did not gain weight over time and had similar PPAR‐γ and ‐α mRNA expression relative to RV dams. Food intake and muscle PPAR‐α /β expression was not affected by any dietary treatment during pregnancy. In conclusion, high vitamin content, perhaps due to folate alone, in the gestational diet increases the obesogenic phenotype of the mothers in later life.

Grant Funding Source: Supported by Canadian Institutes of Health Research, Institue of Nutrition, Metabolism and Diabetes
Effects of brewed green and black tea on inflammation, apoptosis and oxidation in men with prostate cancer (136.3)
Authors: Henning Susanne , Wang Piwen , Carpenter Catherine , Aronson William , Heber David
Published: June 2021

Preclinical and epidemiologic studies suggest chemopreventive effects of green tea (GT) and black tea (BT) in prostate cancer. We conducted a phase II randomized controlled trial in which 93 men diagnosed with prostate cancer consumed six cups of brewed GT, BT or water (control) daily prior to radical prostatectomy (RP). GT consumption resulted in lower early pro‐apoptotic events (decreased Bcl‐2 staining (p=0.02)) and lower pro‐inflammatory nuclear staining of nuclear factor kappa B in prostatic tumor epithelial tissue compared to water consumption (p=0.03). Tea consumption did not affect proliferation (Ki67) and showed a non‐significant trend to decreased oxidant DNA damage (8OHdG) in the RP tissue. Tea polyphenols were detected in prostate tissue from 32 of 34 men consuming GT but not in the other groups. BT polyphenols (theaflavins) were not found in prostate tissue from men consuming BT and no significant changes in tissue biomarkers were observed in the BT group. Evidence of a systemic antioxidant effect was observed as a decrease in urinary 8OHdG only with GT consumption. GT, but not BT or water, also led to a small but statistically significant decrease in serum prostate‐specific antigen levels. Given the GT‐induced changes in inflammation, oxidation, and early apoptotic events, future studies are warranted to further examine the role of GT for prostate cancer prevention and treatment.

Grant Funding Source: Supported by RO1CA116242 and P50CA092131
Pre‐diagnostic plasma unmetabolized folic acid and the risk of colorectal cancer (136.5)
Authors: Cho Eunyoung , Townsend Mary , Selhub Jacob , Paul Ligi , Rosner Bernard , Fuchs Charles , Willett Walter , Giovannucci Edward
Published: June 2021

Objective: To investigate the association between pre‐diagnostic plasma unmetabolized folic acid levels and risk of colorectal cancer (CRC).

Methods: We evaluated pre‐diagnostic plasma levels of unmetabolized folic acid in relation to the risk of CRC in nested case‐control studies within two prospective cohort studies, Nurses’ Health Study (NHS) and Health Professionals Follow‐up Study (HPFS). Blood samples were collected prior to folic acid fortification (in 1989‐1990 in NHS and 1993‐1995 in HPFS). A total of 618 cases of CRC up to 2010 follow‐up and 1215 matched controls (344/680 women and 274/532 men) were included in the analysis.

Results: Unmetabolized folic acid was detected in 22 % of the controls and 21% of the CRC cases. Among those detected, the median value of unmetabolized folic acid was 0.61 nmol/L in both controls and cases. Overall, pre‐diagnostic plasma levels of unmetabolized folic acid were not associated with CRC risk. Compared to those undetected, the multivariate relative risks (RRs) of CRC were 1.14 (95% confidence interval (CI), 0.79‐1.65) for <0.5 nmol/L and 1.13 (95% CI, 0.79‐1.60; P for trend, 0.30) for 蠅0.5 nmol/L. When we stratified the association by sex, there was a positive association between unmetabolized folic acid levels and CRC risk among men (RR=1.58 [95% CI 1.01‐2.49] for 蠅0.5 nmol/L vs. undetected) but not among women (RR=0.64 [95% CI 0.39‐ 1.05] for the same comparison; P for interaction=0.05). The association was also positive among those with methylene‐tetrahydrofolate reductase (MTHFR) CT/TT genotype (RR=2.20 [95% CI 1.24‐3.90] for 蠅0.5 nmol/L vs. undetected) but not among those with CC genotype (RR=0.49 [95% CI 0.22‐1.08] for the same comparison; P for interaction=0.03) Conclusions: Pre‐diagnostic plasma levels of unmetabolized folic acid from pre‐fortification period were not associated with the risk of CRC. The positive association in some subgroups of the population needs to be further explored.

Grant Funding Source: Supported by CA136950, CA87969, and CA55075
Resolvin D1 protects from neutrophil mediated joint destruction in inflammatory arthritis (146.5)
Authors: Norling Lucy , Headland Sarah , Dalli Jesmond , Serhan Charles , Perretti Mauro
Published: June 2021

Dietary supplementation with omega‐3 PUFA is efficacious in reducing joint pain, morning stiffness and NSAID usage in rheumatoid arthritis patients. Yet there is an unmet need to determine how they exert beneficial effects. Recent discoveries have determined that omega‐3 PUFA can be enzymatically converted in vivo to novel pro‐resolving bioactive lipid mediators, termed resolvins. We recently showed that resolvin D1 (RvD1) limits PMN recruitment in experimental peritonitis via its receptor ALX (PMID:22499990), yet whether RvD1 exhibits anti‐arthritic effects remains to be determined. In an inflammatory arthritis model; mice were administered veh (0.1% EtOH, i.p.) or treated with 17R‐RvD1 (100ng, i.p. daily) either prophylactically (day 0) or therapeutically (ensuing overt signs of arthritis; day 4), and graded for arthritis severity (max score 12). Prophylactic treatment with RvD1 significantly lowered the clinical score and hind paw oedema, and attenuated leukocyte infiltrate and prostanoids within the paw. Importantly, therapeutic treatment reduced the peak in arthritis severity and shortened the resolution interval. Protective actions of RvD1 were abolished in ALX (fpr2/3) null mice, indicating the crucial role of this GPCR in RvD1 joint protective actions. These studies indicate that the pro‐resolving mediator RvD1 offers a valuable therapeutic approach for inflammatory joint diseases including rheumatoid arthritis.

Grant Funding Source: Supported by Arthritis Research UK Fellowship 19909 to LVN and The Wellcome Trust 086867/Z/08/Z
The role of prion protein sequence in cross‐species prion transmission (153.2)
Authors: Kurt Timothy , Bett Cyrus , Fernández‐Borges Natalia , Jiang Lin , Eisenberg David , Castilla Joaquin , Wüthrich Kurt , Aguzzi Adriano , Sigurdson Christina
Published: June 2021

Prion diseases are fatal neurodegenerative diseases caused by the conversion of normal cellular prion protein, PrPC, to a misfolded isoform known as PrPSc. Zoonotic prion transmission was reported during the bovine spongiform encephalopathy (BSE) epidemic, yet assessing the risk of cross‐species prion transmission remains challenging. We and others have studied how specific amino acid residue differences between species impact prion conversion, and have found the β2‐α2 loop region of the prion protein (residues 165‐175) markedly influences transmission. Here we examined the impact of amino acid substitutions within the loop on cross‐species prion transmission in vivo and in vitro. We found that transgenic mice expressing β2‐α2 loop substitutions completely resisted infection with two strains of mouse prions and with deer CWD prions. To investigate the impact of individual amino acid residues on conversion, we used an in vitro conversion assay and found that substitutions at certain residues within the loop, such as position 169, dramatically reduced conversion whereas other substitutions had no effect. We propose a model based on steric zipper interactions to explain these results. These studies indicate that the β2‐α2 loop modulates cross‐species prion transmission and suggest this region as a target for rational therapeutic design.

Grant Funding Source: NS055116, NS069566, and U54AI0359
Development of the hyoid‐larynx complex and its variants: significance for clinical and forensic studies (17.7)
Authors: de Bakker Bernadette , de Bakker Henri , Soerdjbalie‐Maikoe Vidija , Moorman Antoon
Published: June 2021

Anatomical variations of the hyoid‐larynx complex occur in 4‐28% of the general population. The embryonic development of this complex, however, has been poorly described, and it is unclear whether current descriptions, which form the basis for the explanation of the development of these variants, are based on observations of original human embryos or on comparative embryology.

To obtain more insight in the development of the hyoid‐larynx complex, we analyzed histological sections of human embryos from the Carnegie Collection. Three‐dimensional models were prepared in a novel interactive portable format providing crucial spatial information and facilitating interpretation. Furthermore we analyzed a forensic‐radiological database with 300 dedicated scans of the hyoid‐larynx explantates.

In contrast to current theories, we observed that the body of the hyoid bone originates from a single growth center, without overt contributions from the second and third pharyngeal arch cartilages. The thyroid and cricoid cartilages develop from mesenchymal packaging, instead of pharyngeal arch cartilages. Most variants are not important in a clinical setting, however they are important in forensic setting if a fracture of the hyoid bone is suspected. In those cases it is of utmost importance to discern between a fracture of the hyoid bone or the thyroid horns and malfusion of those parts.
Is anatomy dissection effective IPE? (18.2)
Authors: Wainman Bruce , Palombella Andrew , Fernandes Alisha , Salfi Jenn
Published: June 2021

Interprofessional education (IPE) within health care programs has been shown to improve patient care and satisfaction, reduce clinical error rates, improve collaborative team behaviour, and diminish negative professional stereotypes. In recognizing this need for IPE as well as the universal commonality and interest in gross anatomy, an IP cadaveric dissection elective was instituted at McMaster University in 2008 and run annually. This 10 week, problem‐based learning gross anatomy elective was carried out by IP teams consisting of students in medical, midwifery, nursing, physician assistant, occupational therapy and physiotherapy programs. Of 100‐140 interested students, 28 are randomly selected and allocated into 4 IP groups consisting of 4‐6 health professions each. Pre‐experience and post‐experience surveys, consisting of the revised Interdisciplinary Education Perception Scale (IEPS) and revised Readiness for Interprofessional Learning Scale (RIPLS), were used to measure changes in attitudes and perceptions towards IPE and collaboration, while qualitative analysis of exit interview data were used to evaluate the entire event. Even though the students volunteered because of interest in an IP event, significant improvements were seen in the RIPL Subscale “Positive Professional Identity” and the IEP Subscale “Competency & Autonomy” and qualitative analysis indicated improvements in role clarity, anatomy knowledge, interpersonal facilitation and IP learning. Qualitative analysis of interview data supports the quantitative findings and highlighted key differences between short and long duration IPE experiences.
Genetic control of upper airway patterning (207.2)
Authors: Sun Xin , Hines Elizabeth , Jones Mary‐Kayt , Verheyden Jamie , Harvey Julie
Published: June 2021

In the trachea and bronchi of the mouse, airway smooth muscle (SM) and cartilage are localized to complementary domains surrounding the airway epithelium. Proper juxtaposition of these tissues produces a balance of elasticity and rigidity that is critical for effective air passage. It is unknown how this tissue complementation is established during development. Here we will present data where we dissected the developmental relationship of these tissues by genetically disrupting either SM formation (through Srf inactivation) or cartilage formation (through Sox9 inactivation), and assessed the impact on the remaining tissue. We found that in both the trachea and main bronchi, loss of one lineage led to an increase in cell number of the remaining lineage. However, only in the main bronchi, but not in the trachea, did the loss of one lineage led to a spatial expansion of the remaining lineage. In addition to SM defects, cartilage‐deficient tracheas displayed epithelial phenotypes, including decreased basal cell number, precocious Clara cell differentiation, and increased secretoglobin expression. Our results demonstrate that the development of upper airway tissues is closely coordinated. A disruption in the formation of one lineage may have widespread consequences on the collective function of these tissues.
A mass spectrometry‐based proteomics approach to identify novel host proteins in enteropathogenic E. coli pedestals (214.3)
Authors: Law H.T. , Chua Michael , Hipolito Kevin , Foster Leonard , Guttman Julian
Published: June 2021

The human intestinal pathogen enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhea and early childhood mortality, particularly in developing countries. A hallmark of these infections is the formation of pedestal‐like morphological structures at sites where EPEC attaches to the host cell surface. Despite their known presence during these infections, their precise role in the disease process remains uncertain. Protein components that have been confirmed at pedestals are limited. Consequently, we hypothesize that undiscovered host proteins are present within EPEC pedestals and that some may play crucial roles in the pathogenesis of EPEC. To test our hypothesis, we developed a novel technique that can separate and concentrate EPEC pedestals from epithelial cells. We then utilized mass spectrometry (proteomics) to identify the protein constituents within these structures. From our screen, we identified 68 proteins with high confidence scores. Among these proteins, 24 corresponded to known and 44 are considered novel pedestal proteins. We have confirmed 9 novel pedestal proteins and have demonstrated the underlying molecular mechanisms in which they are hijacked by EPEC.

Grant Funding Source: NSERC
Breastmilk stem cell transfer from mother to neonatal organs (216.4)
Authors: Hassiotou Foteini , Heath Blake , Ocal Oz , Filgueira Luis , Geddes Donna , Hartmann Peter , Wilkie Thomas
Published: June 2021

Human breastmilk contains stem cells that express pluripotency genes and differentiate in culture to cells from all three germ layers. The fate of these stem cells in the breastfed infant is unknown. We used a mouse model to test tissue distribution and latency of milk stem cells in the neonate. Wild‐type pups were cross‐fostered by mothers that ubiquitously express TdTomato (TdT). Live imaging of organs from TdT‐foster pups within the first 3 weeks after birth revealed the presence of milk‐derived TdT+ cells in the stomach and thymus but not in wild‐type controls nursed by their biological mothers. Confocal imaging of immuno‐stained organs of the TdT‐fostered pups revealed TdT+ cells in stomach and liver. TdT+ cells in the gastric lumen adhered to and penetrated the stomach wall. Some of these TdT+ cells expressed stem cell markers OCT4, NANOG and CD49f. Thymus harbored clones of TdT+ milk‐derived cells expressing the stem cell markers. In close proximity to the double positive population we observed TdT+ milk‐derived cells with low or no expression of the stem cell markers, suggesting differentiation and assimilation with host tissue and/or homing of other types of TdT+ milk cells in these organs. The above now demonstrate that as in humans, mouse milk contains cells that express stem cell markers. These studies in mice provide the first evidence of migration and integration of breastmilk stem cells to organs of the neonate.
Wound healing of cutaneous nitrogen mustard injuries by dynamic modulation of gap junction communication using connexin43 antisense oligodeoxynucleotides (216.6)
Authors: Chang Yoke‐chen , Wang James , Hahn Rita , Chang Hui‐Ying , Gordon Marion , Gerecke Donald
Published: June 2021

Gap junction communication is tightly regulated in skin wound healing. Studies showed that targeting of connexin43 (Cx43) improves skin wound repair in diabetes. Vesicant induced skin injury results in dermal‐epidermal junction disruption and delayed wound repair. We treated Cx43 antisense oligodeoxynucleotides (asODN) on NM exposed SKH‐1 mouse dorsal skin to evaluate the wound healing response. We examined markers of basal keratinocytes, keratin5 (K5); epidermal differentiation, keratin10 (K10); proliferation, Ki67; and basement membrane, laminin 332 (LM332). Immunofluorescent (IF) studies showed reduced K5 expression, loss of K10 and Ki67, and an interrupted LM332 pattern of expression 1 day after NM exposure. By day 3, K10 reappeared in a diffuse pattern. By day 7, the LM332 appearance was thick and continuous. By day 10, there was intense expression of Ki67, accompanied by extreme hyperplasia. Animals treated with Cx43AsODN after NM exposure had an accelerated wound rate and improved reepithelialization so that the skin marker patterns above appeared at earlier timepoints. RT‐PCR and Western blot analysis showed significant reduction of Cx43 at days 3 and 7. IF studies also showed that Cx43 asODN treatment successfully reduced Cx43 expression and modulated higher Cx26 expression in the wound margin by day 3. The use of Cx43asODN may alter Cx expression and accelerate vesicant wound repair.

Grant Funding Source: Supported by ES005022, EY09056, and NIAMS U54AR055073
Dynamic regulation of cleavage and polyadenylation in differentiation and development (225.3)
Authors: Tian Bin
Published: June 2021

Transcripts of most mammalian genes display alternative cleavage and polyadenylation (APA). Sites of APA (pAs) located in the 3’‐most exon typically lead to mRNA isoforms with different 3’ untranslated regions (3’UTRs), and those located in upstream introns or exons also cause different coding sequences. We previously reported a general trend of relative upregulation of promoter‐distal pA isoforms during cell differentiation and embryonic development. Here we examined different cell differentiation systems with 3’READS, a recently developed deep sequencing method to analyze APA isoform expression. We identified cell type‐specific, and differentiation‐specific APA events, and examined ontologies of genes regulated at the APA level. We analyzed how APA is related to overall gene expression. We have found that APA is an important layer of gene regulation, impacting cell identity and differentiation state. In addition, to understand the role of core cleavage and polyadenylation factors (called C/P factors) in APA, we carried out a global knockdown of expression of C/P factors and examined how APA changes in response to C/P factor perturbation is related to APA regulation in cell differentiation.
Miro1‐directed mitochondrial movement protects against neurodegeneration in mice (234.3)
Authors: Shaw Janet , Nguyen Tammy , Oh Sang , Weaver David , Maxfield Dane , Lewandowska Agnieszka , Macfarlane Jane , Saunders Gerald , Koshiba Takumi , Feldman Eva , Hajnóczky Gyorgy
Published: June 2021

Aberrant mitochondrial distribution is observed in numerous neurodegenerative disorders. A popular, but untested, model proposes that local energy deprivation resulting from mitochondrial movement and distribution defects causes the neuronal degeneration associated with these diseases. The Mitochondrial Rho GTPase (Miro) is the only known receptor specific for mitochondrial movement. Mammals ubiquitously express two 60% identical orthologs of Miro (Miro1 and Miro2), but it is unknown whether these orthologs have overlapping or distinct functions. Using two new mouse models lacking a Miro1 conditional allele, we show that: 1) Miro1 has a more specific physiological role than expected, 2) Miro1 has surprisingly specific functions in mitochondrial movement and neuron development/maintenance, and 3) Disruption of Miro1‐mediated mitochondrial movement does not compromise mitochondrial respiration. Thus, defects in mitochondrial distribution are sufficient to cause neuronal deficits in mammals.
A commercialized dietary supplement alleviates joint pain in community adults: a double‐blind, placebo‐controlled community trial (245.8)
Authors: Nieman David , Shanely R. Andrew , Luo Beibei , Dew Dustin , Meaney Mary Pat , Sha Wei
Published: June 2021

InstaflexTM is a joint pain supplement containing glucosamine sulfate, methylsufonlylmethane (MSM), white willow bark extract (standardized to 15% salicin), ginger root concentrate, boswella serrata extract (standardized to 65% boswellic acid), turmeric root extract, cayenne, and hyaluronic acid. Subjects included 100 men and women, ages 50‐75 years, with a history (>3 months) of joint pain, and were randomized to Instaflex or placebo (3 colored gel capsules per day for 8 weeks, double‐blind administration). Joint pain severity was significantly reduced in Instaflex compared to placebo (8‐week WOMAC, ‐37% versus ‐16%, respectively, interaction effect P=0.025), with group differences emerging by week 4. Improvements in ability to perform daily activities and stiffness scores in Instaflex were most evident for the 74% of subjects reporting knee pain (8‐week WOMAC function score, ‐39% versus ‐14%, respectively, interaction effect P=0.027; stiffness score, ‐30% versus ‐12%, respectively, interaction effect P=0.081). Patterns of change in SF‐36, systemic inflammation biomarkers, and the 6‐minute walk test did not differ significantly between groups. Results from this randomized, double blind, placebo‐controlled community trial support the use of the Instaflex dietary supplement in alleviating joint pain severity in middle‐aged and older adults.
Dietary DHA supports glucose use by muscle in association with improved endocannabinoid system gene expression in C57/blk6 mice (248.7)
Authors: Kim Jeffrey , Pedersen Theresa , Newman John , Watkins Bruce
Published: June 2021

Elevated levels of endocannabinoid system (ECS) ligands have been associated with increased adiposity and obesity, and are likely modulators of type II diabetes pathogenesis. High CB1 receptor expression and circulating arachidonic acid (AA)‐derived endocannabinoids (EC) have been found in both obese and diabetic patients. We wanted to determine whether dietary docosahexaenoic acid (DHA) could influence endocannabinoid tone, which includes the action of ligands, receptors, and EC synthesis and degradation enzymes, to prevent an overactive ECS whilst improving aspects of euglycemia. To assess this, male C57bl/6 mice were fed a semi‐purified diet containing DHA or control for 118 d. Serum, gastrocnemius, epididymal fat pads, and liver were collected after 62 and 118 d of feeding. DHA diet fed mice had lower epididymal fat pad masses compared to the control diet group. A compensatory response resulted in DHA fed mice with higher EC receptors and synthesis/degradation enzyme mRNA expression. However, AA‐derived EC (2‐AG and AEA), and downstream ECS activation markers were reduced in DHA fed mice at both 62 and 118 d, despite time dependent increases in these EC. Mice fed the DHA diet had higher glucose‐related (GLUT4 & Insulin‐R) mRNA and proteins. Collectively our findings demonstrate that a DHA diet fed to mice resulted in a reduction in ECS activation leading to an increase in glucose uptake markers and lipolysis.
Manganese supplementation increases adiponectin secretion by upregulating disulfide bond A‐like protein in endothelial cells and Zucker diabetic fatty rats (249.1)
Authors: Burlet Elodie , Jain Sushil
Published: June 2021

Adiponectin is an insulin sensitizer that plays a role in the regulation of lipid and glucose homeostasis and has anti atherosclerotic effects. It is synthesized as a single polypeptide and is then assembled in the endoplasmic reticulum into different multimers. An adiponectin‐interacting protein named disulfide bond A‐like protein (DsbA‐L) was recently found to play an important role in multimerization of adiponectin; impairment in DsbA‐L is associated with diabetes and low adiponectin levels. Manganese (Mn) levels have been found to be low in atherosclerotic and diabetic patients. Previous studies in our lab have shown that Mn supplementation can lower blood cholesterol and ICAM‐1 levels in ZDF rats and reduce MCP‐1 secretion and monocyte adhesion to endothelial cells in HUVECs model (J. Biol. Chem, 2013 Mar 1; 288 (9):6409‐16). This study investigates the hypothesis that Mn supplementation affects DsbA‐L expression levels, and thereby adiponectin levels, in ZDF rats and endothelial cells (HUVECs). Results show that diabetic rats supplemented with Mn (gavaged daily with 16 mg/kg BW for seven weeks, n=5) have higher plasma adiponectin levels (41%, p=0.007) and higher DsbA‐L expression in the liver compared to control diabetic rats (n=6). Similarly, pre‐treatment with Mn (with 5 or 10 μM for 24 h) prevented the decrease in DsbAL expression and adiponectin levels in HUVECs and 3T3L1 adipocyte cells exposed to high glucose and MCP‐1. We further investigated the role of DsbA‐L protein by knocking down its expression in HUVECs. Preliminary studies with DsbA‐L siRNA show that beneficial effects of Mn supplementation on NOX4 expression and other biomarkers of vascular inflammation were abolished in DsbA‐L knocked down cells. Thus, this study demonstrates that DsbA‐L mediates the beneficial effects of Mn on adiponectin and provides a novel mechanism by which Mn supplementation reduces biomarkers of vascular inflammation in diabetes.
A whole grain diet reduces blood pressure in overweight/obese adults 151; a randomized control trial (249.3)
Authors: Kirwan John , Malin Steven , Scelsi Amanda , Louis‐Kullman Emily , Haus Jacob , Kochhar Sunil , Godin Jean‐Philippe , Ross Alastair
Published: June 2021

Epidemiological studies suggest that dietary whole grain intake is associated with a lower incidence of cardiovascular disease. However, there are no Level‐1 data from randomized control trials (RCT) to support these observations. We performed a double‐blind crossover RCT in 40 sedentary overweight/obese (39.1±1.1 years, BMI: 32.5±0.7 kg/m2) adults to test the hypothesis that a whole grain diet reduces cardiovascular risk. Diets were provided based on meeting nutritional guidelines for fat, protein and carbohydrate (i.e. ~28, 18, and 54%) and contained either whole grain (50 g/1000 kcal), or refined grain. Each diet was provided for 8 weeks with an 8‐10 week washout period between diets. A total of 33 subjects completed the study. Mean energy intake, based on resting metabolic rate and a sedentary lifestyle activity factor, was similar between the whole grain and refined grain diets (2051.4±60.7 and 2032.8±60.1 kcal/d, P=0.46). Diets were also similar in macronutrient intake, saturated fat, dietary cholesterol, sodium, and potassium. Compared to baseline, the whole grain and refined grain diets induced similar weight/fat loss (~2 kg, P<0.05). Both diets reduced cholesterol, LDL cholesterol and triglycerides (all P<0.05), but insulin was only reduced after the whole grain diet (P<0.05). Although both diets reduced systolic blood pressure by ~3.5 mmHg, the improvement in diastolic blood pressure (‐5.9±1.1 vs. ‐2.3±1.4 mmHg, P<0.05) and pulse pressure (2.6±2.1 vs. ‐2.5±1.6 mmHg, P<0.05) was significantly greater after the whole grain compared to the refined grain diet. We conclude that a whole grain diet reduces cardiovascular risk factors and produces a clinically significant reduction in diastolic blood pressure in overweight/obese adults independent of weight loss. Since diastolic blood pressure is a predictor of mortality in adults under the age of 50 years, increased whole grain intake may provide a protective nutritional prevention and treatment strategy for hypertension.

Grant Funding Source: A grant‐in‐aid from Nestle and NIH UL1‐RR024989
Tracking healthy behaviors in Head Start children (251.7)
Authors: Swanson Ann , Craven Tara , Gibson Cheryl , Harvey Susan
Published: June 2021

Background. Overweight among preschoolers remains a public health concern, especially in low‐income children. Methods. To encourage healthy behaviors, Head Start families participated in a 6‐week program to promote dietary and physical activity habits. Families tracked their behavior weekly as it related to 5 specific goals: 1) eat more fruits and vegetables, 2) eat more low‐fat dairy, 3) drink less soda, 4) eat less fast food, and 5) be more active. To gain an understanding of the benefits and challenges of the program, site directors and teaching staff were asked to complete a brief survey. Results. Five Head Start programs participated with 230 families completing weekly data tracking forms. Significant increases were reported in servings of fruits/vegetables (p<.01), low‐fat dairy servings (p<.01), and minutes of physical activity (p<.01). Significant reductions in consumption of soda (p<.01) and fast food (p=.01) were also found. Ninety percent of respondents (n=79) indicated the tracking calendars and weekly newsletters motivated families to improve the targeted health behaviors. Most respondents (87%) indicated the distribution of weekly calendars and newsletters to families was easy. When asked what parents were sharing about the program, comments included how the program helped parents become more aware of unhealthy choices, how children were excited to participate, and the barriers families faced while trying to eat healthier. Recommendations included the need to include family‐based nutritional classes and additional resources. Discussion. Based on positive findings, Head Start programs should continue to collaborate with families to promote healthy behaviors within the home setting.
Three doses of vitamin D on bone mineral density in older women: a pilot study (257.4)
Authors: Pop Liliana , Sukumar Deeptha , Schlussel Yvette , Schneider Steve , Stahl Theodore , Gordon Christopher , Shapses Sue
Published: June 2021

Reports show that higher doses of vitamin D (vitD) intake than the recommended have no beneficial effect on BMD. In addition, both aging and weight loss reduce BMD. Our goal here is to determine whether vitD affects bone over 1 year in postmenopausal women who were counseled for weight control. Design: Fifty‐eight obese/overweight women (age, 58 ± 6 years; body mass index, 30.1 ± 3.8 kg/m2, serum 25‐hydroxyvitamin D (25OHD), 27.1 ± 4.4 ng/ml), were randomly assigned to one of 3 doses of vitD (A, 600 IU/d; B, 2000 IU/d; C, 4000 IU/d) with 1.2 Ca g/d. We analyzed 25OHD, parathyroid hormone and BMD and strength parameters by dual energy x‐ray absorptiometry and peripheral quantitative computed tomography. Results: At 1 year, serum 25OHD levels differed between groups (p < 0.01) and increased to 30.4 ± 5.2, 35.8 ± 4.5 and 41.5 ± 6.9 ng/ml, in groups A, B and C, respectively. Weight change was similar between groups (‐3.0 ± 4.1%). There was no significant BMD changes from baseline at the hip, spine or radius, but there was a decrease (p < 0.05) for total volumetric bone mineral content (BMC) (‐0.9 ± 3.4%), trabecular BMD (‐1.3 ± 3.8%) and BMC (‐2.9 ± 9.9%). Also, vitD affected cortical thickness over time (p<0.05) showing a change of ‐0.07± 0.22, +0.02 ± 0.15 and +0.09 ± 0.21 mm, for groups A, B and C, respectively. In conclusion, the vitD dose‐response on serum 25OHD levels improves cortical thickness but does not affect BMD or strength parameters.

Grant Funding Source: NIH‐AJ12161
Vitamin D deficiency is associated with anemia: a cross‐sectional study in healthy adults in the United States (260.3)
Authors: Smith Ellen , Alvarez Jessica , Martin Greg , Zughaier Susu , Ziegler Thomas , Tangpricha Vin
Published: June 2021

Objective: We aimed to examine the relationship between vitamin D status and anemia, hypothesizing that lower vitamin D status would be associated with increased risk for anemia, particularly anemia of inflammation.

Methods: We conducted a cross‐sectional analysis of the Emory/Georgia Tech Predictive Health cohort (n=638), a generally healthy adult cohort. Anemia was assessed via hemoglobin concentration. Multivariable logistic regression was used to evaluate the association of vitamin D status with anemia, controlling for significant covariates of anemia. Relationships between vitamin D status and subtypes of anemia were assessed.

Results: Vitamin D deficiency (serum 25‐hydroxyvitamin D < 20 ng/mL) was significantly associated with anemia in bivariate analysis (OR: 2.64; 95% CI: 1.43, 4.86). After accounting for an interaction between vitamin D deficiency and race, vitamin D deficiency was significantly associated with anemia in blacks (OR: 6.42; 95% CI: 1.88, 21.99), controlling for anemia covariates, but not in whites. When stratified by subtype of anemia vitamin D deficiency was significantly associated with anemia of inflammation only (OR: 3.11; 95% CI: 1.08, 8.91).

Conclusion: Vitamin D deficiency was significantly associated with anemia, specifically anemia of inflammation in a healthy adult US cohort. Racial differences were observed in the association between vitamin D status and anemia.

Grant Funding Source: Supported by: T32 DK 7734‐17, UL1 TR000454, T32 DK007298‐32S1, K23 AR054334, K24 RR023356.