bluuebunny/crossref_metadata_2025 huggingface dataset:
An Advanced Anatomy Dissection Lab Environment Combining Physical Dissection with Digital Dissector and Real‐Time Video Display
Authors: Khan M. A. , Matz Donald G. , Canby Craig A. , Dyche William J. , Hisley K. Calvin
Published: June 2021

Current trends in the use of clinical medical imaging will demand increasing integration of 2D and 3D visual cadaver dissection media and correlated radiological material into the gross anatomy curricula. In addition, increasing class size and the technical sophistication of incoming students necessitates an efficient and well‐organized delivery of dissection‐based anatomical education.

Des Moines University's gross anatomy lab can accommodate 215 students. Each dissection table is a two‐sided operational unit where one student “navigates” for the other four dissectors by tracking and coordinating their work on a table‐side computer and a wall mounted monitor.

Dissection sessions begin with a live, streaming video demonstration displayed on the student monitors at each dissection table. An instructor uses a cadaver to provide live demonstration of the dissection via an adjacent video control room that contains a camera mounted on a surgical boom. Students are then able to use a server‐based digital dissector with hot‐linked structures already pre‐programmed to appear at the students prompting. They are thus able to have immediate table‐side access to both text and atlas‐based visual images to guide them during their dissection exercises. This design allows the application of a vast range of text and visual materials displayed simultaneously and individually to 40 tables.
Biomechanical gradients and early vasculogenic patterns
Authors: Little Charles D. , Czirók András , Cui Cheng , Zamir Evan A. , Perryn Erica D.
Published: June 2021

Vasculogenesis occurs within constantly expanding tissues. Explanted mouse allantoic endothelial cells (ECs) assemble a vascular network, de novo, when placed in culture. The allantoic network is situated on the upper surface of a mesothelium that expands ten‐fold during the process. Similarly, quail embryos manifest vast tissue expansion during vasculogenesis. Computational time‐lapse imaging allows quantification of primordial EC motility, as well as the motion of surrounding tissue. Allantoic ECs at the base of multicellular sprouts normally move distally “over” existing sprout cells to form new tip‐cells; and loss of VE‐Cadherin activity prevents this motile behavior. Other time‐lapse data suggest that EC sprouts respond to mechanical gradients and favor locomotion along rigid biological structures — such as cords of cells or bundles of ECM. We propose a mechanically‐based morphogenetic field in which primordial ECs initially extend sprouts using integrin‐ECM adhesions. Once formed, however, movement of “new” sprout tip cells is mediated by VE‐cadherin based motility. Certain sprout “base” cells appear to be mechanically stimulated to become motile and move distally to replace existing tip cells — so that advancing vascular sprouts can “keep‐up” with the expanding tissues. Thus, endogenous biomechanical tissue gradients partially govern assembly of vascular networks in warm‐blooded animals.
Regulation, Production, and Distribution of Neo‐organs
Authors: Bertram Timothy A
Published: June 2021

Replacing diseased tissues and organs represents a significant unmet medical need. Limited supply and complications such as chronic metabolic alterations, rejection, and limited product shelf‐life have rendered organ transplantation a costly and risky approach. Autologous organ replacement may be a less risky approach. Neo‐organs and neo‐tissues produced from a patient's own cells ex vivo provide a foundation to harness the body's ability to regenerate. Organ or tissue function is restored by the regenerative healing that follows implantation. These products may fulfill a significant unmet medical need for patients who have partial or complete organ loss. Recent scientific advances in tissue engineering technologies and their use in regenerative medicine have helped us better understand host tissue healing responses. These advances have allowed product development to fit into existing and emerging regulatory guidelines and remain commercially viable. Neo‐organ production requires defined raw materials and a reproducible manufacturing process where the appropriate constituents can be brought together ex vivo to form an implantable neo‐organ product. Advancements in the science, technology, and production of regenerative medical products have revealed fundamental and shared aspects from which generalizations about approaches to development and regulatory pathways can be made.
Structural basis for transcription repair coupling in bacteria
Authors: Andrew Darst Seth , Deaconescu Alexandra , Chambers Anna L , Smith Abigail J , Savery Nigel J , Nickels Bryce E , Hochschild Ann
Published: June 2021

Transcription Repair Coupling Factor (TRCF, the product of the mfd gene) is a widely conserved bacterial protein that couples DNA repair with transcription. TRCF recognizes RNA polymerase (RNAP) stalled at a non‐coding template site of DNA damage, disrupts the transcription complex to release the transcript and enzyme, and recruits the DNA excision repair machinery to the site. The mechanism of RNA release has been illuminated by the discovery that TRCF causes forward translocation of RNAP through an ATP‐dependent motor that is homologous to that of the Holliday branch migration protein RecG. TRCF is a large (130 kDa), multi‐functional protein with a complex structure/function relationship. In a process involving its ATP‐dependent DNA translocase activity, TRCF is able to dissociate the highly stable ternary elongation complex (RNAP/DNA/RNA), resulting in the release of the RNAP and the RNA transcript in a true termination reaction. This is one of only three known mechanisms for transcription termination in bacteria. We will present the 3.2 Å‐resolution X‐ray crystal structure of TRCF, along with models for its interaction with the RNAP elongation complex.
Follow‐up Data on Selective Indicators for Reduction of Cardiovascular Risk Factors among African Americans
Authors: Dawkins Norma L , Pace Ralphenia Diggs
Published: June 2021

Dietary and lifestyle changes are important in reducing morbidity and mortality associated with cardiovascular disease (CVD). In Macon County the death rate from CVD in 2004 was 395 per 100,000 was higher than the state (377.5) as well as the national average (336.6). The objective of this study was to determine the sustained effectiveness of the twelve‐week Tuskegee University Nutrition Outreach Program (TUNOP). After the 3 month program ended, participants were evaluated again six months later.

Biological indicators and food frequency consumption were collected on 58 participants. The six‐month evaluation determined if there was continued adherence to the TUNOP procedure after the 3 month intervention which was designed to decrease CVD risk factors.

Results indicated that lipid values decreased significantly (P≤ 0.01) from 207.79 mg/dl to 176.01mg/dl for total cholesterol and LDL‐C from 122.26mg/dl to 104.53mg/dl (P≤ 0.01). HDL‐C decreased from 59.48 to 53.58 mg/dl, and weight remained constant while blood pressure increased. A significant increase in dietary intake was observed for beans, peas, tofu and soy products. In contrast, significant decreases (P≤ 0.05) were observed with the frequency of consumption of ice cream, meats, sweets, salty snacks and soda pop. Therefore, continued reduction in selective biological and dietary indicators for cardiovascular disease risk suggest that adherence to recommendations for improved diet and lifestyle changes did reduce CVD risk among these participants.
Difference in Stress, Depression, and Self‐esteem along with Physical Activity Levels between Overweight and Obese Perimenopausal Women
Authors: Hwang Mi‐Ja , Hwang Deok‐Sang , Song Mi‐Yeon
Published: June 2021

Physical activity (PA) is known to be beneficial for mental health through decreasing depression and stress. In this study, we investigated the relationship between body composition, stress, depression, and self‐esteem with PA level. International Physical Activity Questionnaire (IPAQ)‐short form, Social Readjustment Rating Scale (SRRS), Stress Response Inventory (SRI), Beck Depression Inventory (BDI), and Self‐Esteem Scale (SES) were administered to perimenopausal women aged 45–55 and BMI≥23kg/m2. The PA level of the total group was 2406 MET‐min/week (n = 42). The PA level of overweight (23 kg/m2 ≤BMI<25 kg/m2) and obese (BMI≥ 25kg/m2) perimenopausal women was 1428 MET‐min/week and 2970 MET‐min/week (p<0.05), respectively. When three levels of PA (inactive, minimally active, and HEPA [Health‐enhancing physical activity] active) were analyzed, all levels consisted primarily of walking activities rather than vigorous activities. Within all groups, BDI, SRRS, and SRI scores were non‐significantly lower and SES was non‐significantly higher in HEPA active group versus other two groups. Anger (subscale of SRI) measures were lower among obese women and SES measures were higher in HEPA active group than others among overweight women (p<0.05). This study shows that the level of PA of Korean perimenopausal women was not insufficient, but the vigorous activity is thought to be necessary to enhance health. Health‐enhancing PA was related with high self‐esteem in overweight women and low anger in obese women. This study was supported by KyungHee University.
Copper is taken up efficiently from albumin and alpha‐2‐macroglobulin by cultured human cells by more than one mechanism
Authors: Moriya Mizue , Ho Yi‐Hsuan , Grana Anne , Nguyen Linh , Alvarez Arissa , Jamil Rita , Ackland Leigh , Michalczyk Agnes , Kim Stephen , Linder Maria C.
Published: June 2021

Normally Cu(II) entering the blood plasma binds tightly to albumin (Alb) and the macroglobulin, transcuprein. However, little is known about how this copper is then taken up by hepatic cells, which are main recipients. Two determine this, the kinetics of copper uptake from these plasma proteins, and effects of inhibitors, were measured in the human hepatic (HepG2) and mammary epithelial (PMC42) cell lines. At physiological concentrations (1–5 uM) both cell types took up Cu from purified human Alb and alpha‐2‐macroglobulin (a2M) at rates similar to each other and to those for Cu‐di‐histidine (His). Uptakes from a2M but not from Alb or His obeyed Michaelis Menten kinetics, with Kms of 4–6 uM. Uptake from a2M but not the others was markedly inhibited by Ag(I) in HepG2 but not PMC42 cells. Copper uptake was not inhibited by Fe(II), Mn(II), or Zn(II) except at very high concentrations (200–500 uM). We conclude that the two plasma proteins mainly responsible for the plasma exchangeable copper pool deliver copper as efficiently as amino acid complexes to mammalian cells; that a2M delivers it primarily to CTR1 in hepatic but not mammary epithelial cells; and that additional as yet unidentified transporters responsible for copper uptake from these plasma proteins remain to be identified. Supported by PHS Grant No. RO1 HD 46949.
Fructan supplementation affects nitrogen partitioning in excreta and stool metabolite concentrations in healthy senior cats
Authors: Barry Kathleen Ann , Hernot David C. , Loo Jan Van , Fahey George C.
Published: June 2021

Inulin‐type fructan (ITF) supplementation was evaluated in senior (≥ 9 y) cats to determine nitrogen (N) partitioning in excreta and stool metabolite concentrations. One of two ITF‐based supplements (oligofructose or SynergyC, an experimental blend of short and long chain ITF) was added to a commercially available wet cat food with low intrinsic fiber content at 1% on a dry weight basis. All cats were fed the diet without supplemental ITF for 7 d, after which cats were assigned to one of three treatment groups for 21 d. Total feces and urine excreted were collected from the cats on d 6–7 (baseline) and d 22–28 (treatment). No differences were observed in food intake; fecal output, dry matter percentage, or score; fecal or urinary ammonia output; or N retention. Supplemental ITF decreased (P < 0.05) N digestibility and fecal histamine, spermidine, and indole concentrations. Supplementation with SynergyC increased (P < 0.10) fecal propionate concentration. While not statistically significant, a numerical decrease in urinary N content was observed in ITF‐supplemented cats. Decreased apparent N digestibility was likely due to an increase in microbial protein synthesis in ITF‐supplemented cats. ITF supplementation may be beneficial to the senior cat as a result of its ability to modulate N concentration in excreta and decrease odor‐ and disease‐causing metabolites in feces.

This work was sponsored by BENEO‐Orafti.
Carotenoid 9', 10'‐monooxygenase: Tumor Suppressor Activity in Prostate Cancer
Authors: Gong Xiaoming , Zaripheh Susan , Xiao Fan , Zarkhin Eli , Buch Vivek P , Magi‐Galluzzi Cristina , Rubin Lewis P
Published: June 2021

Carotenoid cleavage monooxygenases (CMOs) are a class of enzymes that cleave various carotenoids (including carotenes, lycopene) to apocarotenoids and retinoids. Carotenoid 9’,10’‐monooxygenase (CMO2) regulates lycopene metabolism. Since lycopene intake may have preventive and therapeutic effects in prostate cancer, we investigated CMO2 in human normal prostate, cancer tissue and prostate cancer cell lines. CMO2 expression is significantly higher in human normal prostate tissues than that in prostate cancers. In prostate cancer cell lines, CMO2 expression is higher in androgen‐sensitive than in androgen‐resistant cells. Lycopene treatment significantly increases CMO2 expression and reduces cell proliferation in androgen‐sensitive cells, but does not affect CMO2 expression or cell growth in androgen‐resistant cells. Over‐expression of CMO2 in various prostate cancer cell lines decreased proliferation regardless of lycopene co‐treatment or androgen‐responsiveness. Exogenous CMO2 expression reduced NF‐κB activity by interfering with phosphorylation and DNA binding of the p65 and p50 components of the NF‐κB complex. CMO2 over‐expression also increased PPAR and p38 activity and reduced Akt1, ERK2 and JNK2 activity. CMO2 suppression apparently contributes to development of human prostate cancer. We also describe previously unexpected functions for the enzyme CMO2, apparently independent of a role in carotenoid metabolism. Our findings (1) suggest that CMO2 may be a tumor suppressor in prostate cancer and (2) support a role of lycopene in the prevention or treatment of androgen‐sensitive prostate cancer (NIH HD42174, RR18728).
Herbal education is reasonable and necessary for Food Stamp program participants
Authors: Lohse Barbara Ann , Rifkin Robin , Stotts Jodi L , Cooper Rayna G
Published: June 2021

Previous research revealed widespread use of herbals by caregivers and children attending the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC).1This project developed a technology‐based herbal program for WIC clinics and examined participant response. Information about 5 most frequently used herbs 1 (garlic, peppermint, ginger, Echinacea, cranberries) formed the basis of a program delivered by digital photo receiver (DPR). WIC staff and target audience helped develop the herbal DPR program and materials for recruitment and assessment. Prior to planned statewide dissemination, responses from 282 (N=26 male) WIC clients (78.6% < 36 years) attending 2 urban and 2 rural clinics were examined. Only 27.3% had no experience with herbal use for themselves or their children; 47.5% were current herbal users. The herbal program was useful for 98.5% and liked by 93.2%; a take away information card was useful for 93.8%. Food Stamp program (FSP) participation was reported by 37.3%; participation increased to 56.4% when including prior food stamp use. FSP participation was associated (P=.04) with current herbal use; 52.5% of FSP participants used herbals compared to 40.2% for persons never participating in FSP. Child mean herbal use was higher with FSP experience (1.26 ±1.28 vs. 0.82 ± 1.26 herbals; P=.004). A trend for FSP participants to learn new information from the DPR herbal program (82.1% vs. 73.5% for non‐FSP; P=.09) combined with widespread herbal use suggests herbal education for FSP participants is reasonable and necessary.

Funded by the Pennsylvania Nutrition Education Tracks, a part of USDA's Food Stamp Program.
Low birth weight is associated with increased lean body mass in healthy children and adolescents.
Authors: Hoffman Daniel , Toro‐Ramos Tatiana , Dolan Meredith
Published: June 2021

Previous studies have reported that persons born with a low birth weight for gestational age (LBW) are at an increased risk for obesity. However, most studies have relied on the Body Mass Index (BMI, kg/m2) to assess obesity and estimate adiposity. Therefore, to better understand the relationship between body composition and birth weight adjusted for gestational age (BWTadj), we completed a study in which body composition was measured using dual energy x‐ray absorptiometry and birth weight was assessed by maternal recall.

The study sample consisted of 101 children with a mean age of 12.8 years, BWT of 3.3 kg, and gestational age 38 weeks. Using backward linear regression analyses, BWT was found to be inversely associated with LBM, independent of the covariates age, Tanner stage , height2, current body weight (WT), race, and gender. Two models were created with the following main results: 1) LBM = −7.4 + −1.7*BWTadj (p=0.024), independent of covariates (R2=0.94); and 2) LBM adjusted for WT = −9.3 + −3.2*BWTadj (p=0.006), independent of covariates (R2=0.40).

Based on our data, we report that that a LBW is associated with higher LBM relative to body weight and height and may explain previously observed associations between LBW and increased BMI.

Funding was provided by the Busch Biomedical Foundation and in part by NIH/NIA grant number P01 AG10120.
Birth weight, childhood BMI and the risk of stroke in adulthood among 191,163 Danish schoolchildren
Authors: Baker Jennifer Lyn , Olsen Lina W. , Truelsen Thomas , Sørensen Thorkild I.A.
Published: June 2021

Objective: Larger body size in childhood raises the risk of CHD in adulthood. We expanded on this and investigated if birth weight and childhood BMI from the ages of 7–13y were associated with an increased risk of stroke in adulthood.

Methods: Subjects were 97,391 boys and 93,772 girls born from 1936–1976 in the Copenhagen School Health Records Register. BMI values were calculated and transformed to z scores. Stroke events were obtained from Danish Hospital and Cause of Death Registers. Cox regressions were performed.

Results: Among boys, birth weight was associated with stroke in adulthood. Compared to boys in the reference category (3251–3750g) those in the lowest (2000–2750g) had a relative risk (RR) of 1.37 (95%CI: 1.22–1.55) of stroke. BMI at 13y was associated with stroke in boys; compared to those with a z score in the reference category (−0.25–0.25) boys with a z‐score >= 0.75 had a RR of 1.11 (95%CI: 1.02–1.21). Similar results were observed for girls. Adjustment for birth weight strengthened these associations.

Conclusions: Birth weight and childhood BMI have independent associations with stroke in adulthood. These results suggest that although some risk of stroke is already present at birth, childhood body size also matters. As it is amenable to intervention, there still remain preventive possibilities.

Supported by: NIH NRSA 1F32DK070491 (to JLB) & Danish National Research Foundation
The Association Between Acculturation and Obesity in Hispanic Women: Findings from the 2005 California Women's Health Survey
Authors: Banna Jinan Corinne , Kaiser Lucia , Townsend Marilyn
Published: June 2021

Objective: To assess the relation of acculturation with obesity in Hispanic women in California.

Methods: We analyzed data from Hispanic women 18 years of age and older (n=1298) from the 2005 California Women's Health Survey (CWHS).

Results: Participants had a mean body mass index (BMI) of 27.6 ± 5.9; over 50% of the sample was classified as overweight (BMI 25–29.9) or obese (BMI ≥30). There was no association between BMI and language acculturation, nor was there a significant difference in BMI between those born in the U.S. and those born elsewhere. For those born outside of the U.S., however, there was a significant association between BMI and years spent in the U.S. (p<.0001), with those classified as overweight or obese having spent more time in the U.S. Of those who had lived in the U.S. <5 years, 19% had a BMI ≥30, compared to 34% of those who had lived in the U.S. >20 years.

Conclusions: These findings show a positive association between length of time in the U.S. and BMI in foreign‐born Hispanic females. Language use and birthplace, however, were not associated with BMI. The way in which acculturation is measured may affect study results and should be considered. Study findings indicate the need for further investigation of the acculturation process and obesity in order to design appropriate prevention strategies.

Supported by the Jastro Shields Graduate Research Scholarship.
Prenatal and postnatal factors associated with high weight retention in young Mexican women
Authors: Hernandez‐Cordero Sonia , Ramakrishna Usha , Neufeld Lynnette M
Published: June 2021

The objective was to explore prenatal and postnatal factors associated with weight retention (WR) at 3 months postpartum (pp) in Mexican women. Women were recruited as part of a randomized supplementation trial < 26 wks of pregnancy and followed to 3 m pp. We studied 415 women. Weight gain (WG) and weight retention were calculated from recruitment to each follow‐up period, representing an underestimation of true weight change. High WR at 3 m pp was defined as > 2.2 kg. Mean age at recruitment was 27.4±5.8 y, gestational age 20.7±5.7 wks. At 3 m pp, mean WR was −0.1±4.0 kg, and 25% of women had high WR. Nearly 40% of women were overweight (25.0–29.9) and 21% obese (≥30) at 3 m pp. Compared to women with lower WR, women with high WR were younger, had higher WG in the second half of pregnancy, had higher WG from the 1st to 3rd m pp and were more sedentary. In multivariate analysis, every kg of WG during pregnancy and from 1 to 3 mo pp, the risk of having a high WR increases by 43% (95% CI 1.3–1.6) and 66% (95% CI 1.4–1.9), respectively. The risk of having a high WR increased for every hour of sitting in the day by 13% (95% CI 1.001–1.3). The results of this study underscore the importance of appropriate WG during pregnancy and in early pp to avoid excess WR. The Oportunidades program is an ideal venue to promote healthy weight gain and physical activity during and after pregnancy.

Founding: Oportunidades program, holding the rites to the data presented.
Efficacy of vitamin D supplementation in an Antarctic ground analog of space flight
Authors: Smith Scott M. , Gardner Keri , Locke James , Zwart Sara R.
Published: June 2021

Vitamin D is critical for persons with limited exposure to ultraviolet (UV) light, including space travelers. Recent studies indicate that optimal serum 25‐hydroxyvitamin D (25‐OH vit D) should be ≥ 80 nM. This study was designed to define the dose of vitamin D needed to maintain 25‐OH vit D at > 80 nM in persons with limited UV‐B exposure. It was conducted during winter in Antarctica at McMurdo Station, when UV‐B radiation levels are essentially zero. The 66 subjects recruited for this blinded study were randomly divided into 3 groups for vitamin D supplementation: 2000 IU/d (n = 18), 1000 IU/d (n = 18), and 400 IU/d (n = 19). Some subjects (n = 7) did not take supplements or took ones of their own choosing. Blood samples were collected about every 2 mo during the winter. About 5 mo after supplementation started, analysis of samples from subjects (n=9, 5, 11, and 3 for the 2000 IU/d, 1000 IU/d, 400 IU/d, and no pill groups) who completed 4 blood draws showed that 25‐OH vit D increased 63% in the 2000 IU/d group (to 82 ± 28 nM), 61% in the 1000 IU/d group (to 85 ± 36 nM), and 51% in the 400 IU/d group (to 64 ± 13 nM). It decreased 13% (to 35 ± 16 nM) in the group not taking supplements. These data will enable us to provide space crews with evidence‐based recommendations for vitamin D supplementation. The findings also have implications for other persons with limited UV light exposure, including polar workers and the elderly. This study was supported by the NASA Human Research Program.
Ghrelin response in malnourished African children undergoing nutritional rehabilitation
Authors: Nweneka Chidi Victor , Sharma Amita , Darboe Momodou K , Doherty Conor P , Cummings David E , Prentice Andrew M
Published: June 2021

Recovery from severe protein energy malnutrition (PEM) requires a strong anabolic drive. We investigated the response of ghrelin, an orexigenic hormone, in malnourished Gambian children undergoing nutritional rehabilitation. Pre‐ and post‐prandial serum ghrelin levels were measured in undernourished cases (children 1–3y, Weight‐for‐Height Z score, WHZ <−2SD) at days 0, 10 and 30 after starting nutritional rehabilitation, and in controls (children seen at MRC Keneba clinic during the same period, matched for age and sex with WHZ >−2,) at day 0. H. pylori (HP) status was assessed using 13C‐Urea Breath Test. Post‐prandial day 0 ghrelin levels were significantly different from the pre‐prandial day 0 (baseline) levels in cases and controls (see Figure). Although not statistically significant, the baseline serum ghrelin was higher in malnourished children (2437pg/ml vs. 1978pg/ml, P=0.2), and fell towards control levels with nutritional rehabilitation. Baseline ghrelin was related to age (cases: r = −0.6, P=0.009; controls: r = −0.6, P=0.005), but not to baseline WHZ, Weight‐for‐Age and Height‐for‐Age Z scores, insulin, hGH or CRP levels. Eradication of HP in 3 subjects led to a fall in ghrelin levels from 3027 pg/ml at day 0 to 947pg/ml at day 30 (P=0.001). The data suggest that normal ghrelin response remains intact in malnourished African children and may be stimulated by HP infection. This study was sponsored by MRC Keneba, The Gambia
Eating rate and satiation with a caloric beverage
Authors: Andrade Ana M. , Dwiggins Jessie , Melanson Kathleen J.
Published: June 2021

We have shown enhanced satiation with slow eating. However, subjects drank more water when eating slowly. Thus, we examined the influence of eating rate on satiation when a caloric beverage is offered. In a randomized design, quick (Q) and slow (S) eating rates were compared in 25 females (20.2±4.8y; BMI=21.3±2.2). After a morning of controlled food and beverage intake and activity, they consumed an ad libitum mixed‐macronutrient lunch and apple juice. Hunger (H), satiety, desire‐to‐eat (DTE), and thirst (T) were rated on visual analogue scales every 5min up to 30min, at 45 & 60min. Q and S conditions were compared by paired t‐tests. Meal durations and eating rates were 8±3min & 27±8min, 106±37Kcal/min & 30±9Kcal/min during the Q & S conditions, respectively (P<0.001 for both). S eating decreased food intake (Q=674±140Kcal; S=598±152Kcal; P<0.05) and increased beverage intake as compared to Q eating (Q=128±59Kcal; S=176±86Kcal; P<0.01). However, no differences occurred in total energy consumed at lunch (Q=802±177Kcal; S=774±195Kcal; P>0.05). At 45 & 60min H and DTE were higher in the Q condition (P<0.05). T was lower at 25, 30, 45 & 60min for the S condition (P<0.05). Regardless of the beverage served, eating slowly can effectively decrease food intake, but it tends to increase beverage consumption. Despite similar ad libitum total energy intake, lower H & DTE scores may support a role of slow eating in satiety.
Development of population‐specific anthropometric prediction equations for children in Brazil
Authors: Toro‐Ramos Tatiana , Rondo Patricia , Sawaya Ana Lydia , Hoffman Daniel
Published: June 2021

The double burden of obesity and underweight is a serious public health dilemma in many developing countries. While anthropometric prediction equations (PE) for fat mass (FM) in children exist, most have been developed in cohorts from developed populations, not countries experiencing the “double burden”. Therefore, using data from 145 pre‐pubertal children (9.8 ± 1.3 yrs) from São Paulo, Brazil, we developed a PE for FM using body composition measured by dual energy X‐ray absorptiometry and skinfold and anthropometric measurements from six sites. The new PE was developed using backward linear regression analysis with weight, triceps, height, gender, and age as independent variables from 30 randomly selected children (R2=0.77). When cross‐validated in 125 children, the population‐specific PE met the criteria for validity (i.e. no significant difference between the means and slope =1 with an X‐intercept = 0) (R2=0.80), while standard, child‐specific PE for FM by Slaughter (R2=0.51), Goran (R2=0.67), and Huang (R2=0.33 and R2=0.23) did not meet these criteria. In conclusion, the newly developed, population‐specific PE appears to be more valid for use in children from developing countries than the standard equations. The use of a ‐specific PE can enhance the assessment of body composition of children in countries experiencing the double‐burden of disease.
Vitamin D is inversely related to parathyroid hormone in newborn Canadian infants
Authors: Grace Trussler Kathryn , Weiler Hope
Published: June 2021

Concentrations of 25(OH)D in the order of 75 nmol/L associated with a plateau in PTH are widely accepted as a target for defining optimal vitamin D status in adults and adolescents. The primary objective of this study was to examine the relationship between serum 25(OH)D and PTH from infant cord blood samples. Cord blood samples from 64 infants were measured for 25(OH)D (Octeia ELISA, Immunodiagnostic systems Ltd, UK) and PTH (Human Bioactive PTH 1–84 ELISA, Immutopics, USA). PTH and 25OHD were inversely correlated using Pearson analysis (r=−0.31 p=0.007, n=62) (Figure one). Infants with 25(OH)D ≤30 nmol/L had PTH values that were significantly higher than those with 25(OH)D 30–50 nmol/L (p<0.05) and ≥50 nmol/L (p<0.01) using ANOVA and Tukey's tests. The data suggests a plateau in PTH at [25(OH)D] 75 nmol/L, but requires further statistical modeling using a larger sample of concentrations above 75 nmol/L. Finding an optimal target for vitamin D status will not only aid in diagnostics but will assist current research on vitamin D supplementation levels for breastfed infants. Research was funded by Canadian Institute of Health Research
Commensal bacteria promote intestinal epithelial restitution by regulating FAK phosphorylation
Authors: Kumar Amrita , Babbin Brian A. , Nusrat Asma , Neish Andrew S.
Published: June 2021

Commensal enteric bacteria, which are often used as probiotics, positively influence epithelial barrier recovery after injury. Restoration of epithelial barrier following injury (restitution) requires intestinal epithelial cell (IEC) migration‐ a process that is dependent on dynamic turnover of focal cell‐matrix adhesions (FAC). The mechanism(s) by which commensal bacteria regulate IEC migration and restitution after injury are however not understood. Thus, our objectives were to explore the signaling pathways by which commensal bacteria influence IEC migration and restitution. We show that in vitro interaction of model IECs with the commensal bacterial strain Lactobacillus rhamnosus, resulted in a significant increase in cell adhesion, spreading and wound closure. This was associated with phosphorylation/activation of key FAC proteins, focal adhesion kinase (FAK) and paxillin within 5 minutes of bacterial contact. Furthermore, bacterial colonization of cultured epithelial cells resulted in rapid and reversible generation of reactive oxygen species (ROS), which induced sustained FAK activation by oxidative inactivation of a FAK phosphatase, PTEN. These observations demonstrate that commensal/probiotic bacteria facilitate epithelial barrier recovery by stimulating ROS, FAK phosphorylation and IEC restitution. This research was supported by a NIH grant AI064462 and DK55679.
Matrix Metalloproteinase (MMP)‐1 Induces Migration and Proliferation of Lung Epithelial Cells
Authors: Herrera Iliana , Cisneros Jose , Ramírez Remedios , Garcia Jorge , Selman Moises , Pardo Annie
Published: June 2021

MMP1 is a multifunctional protease that cleaves fibrillar collagens and process a number of non‐matrix components. Studies in our laboratory have demonstrated that MMP1 is upregulated in idiopathic pulmonary fibrosis. However, the protein is expressed in alveolar epithelial cells and it is virtually absent in the lung interstitium and fibroblastic foci where collagens are being accumulated. In this study we explored the putative function of epithelial expression of this enzyme. Mouse lung epithelial cells (MLE‐12) were transfected with human MMP1 linked to the expression vector pQCXIP. Control cells were transfected with the vector. Cell migration was analyzed using Boyden chambers covered with collagen and by in vitro wound healing assay using epidermal growth factor as chemoattractant. Cell proliferation was evaluated with WST‐1 reagent. MMP1 transfected epithelial cells showed a significant increase in cell migration (DO: 0.39±0.4 vs 0.24±0.2; p<0.05). In the wound healing assay MMP1 transfected cells covered 100% of the wound at 36 h, compared with 60% of the control epithelial cells. Likewise, MMP1 transfected cells exhibited a significant increase of proliferation at 36 h (300±20% vs 254±11%; p<0.05) and at 48 h (394±33% vs 300±11%; p<0.01). These findings suggest that MMP1 may have a role in lung epithelial cell migration and proliferation.

Funded by: UNAM, Grant SD1.PTID.05.6
Repopulation of human hepatocytes in immunodeficient mice
Authors: Liu Chen , Witek Rafal , Bess Jennifer , Zhu Haizhen , Lin Philip
Published: June 2021

A robust humanized mouse model can be a valuable experimental system for variety of studies on human diseases. Several reports have demonstrated that human hepatocytes can repopulate in the liver of immunodeficient (scid) and urokinase plasminogen activator (uPA) transgenic mice (scid/Alb‐uPA). The technical challenge limits the broad application of this model. Here we developed a relatively simple model for human hepatocyte repopulation. Rag2−/− mice were treated with monocrotaline (MCT) to inactivate the endogenous hepatocyte proliferation. After two injection at 2‐week interval, partial hepatectomy was performed. At the time of hepatectomy, 2 million freshly isolated human hepatocytes were injected into the spleen. Serum samples and liver tissue were collected for analysis of human liver protein markers (i.e., albumin and alpha‐1 antitrypsin). Our results suggest that human hepatocytes are repopulated in MCT‐treated Rag2−/− mice. The repopulation efficiency is between 2% to 10%. The human hepatocytes persist in the mouse liver for at least three months. Current experiments aim at improving the repopulation efficiency and infecting the mice with human hepatitis C virus (HCV). We believe this model would facilitate investigations of a number of human liver diseases. The work was partially supported by NIH DK02958 and AI064357
DIAGNOSIS OF PULMONARY ASPERGILLOSIS UTILIZING PROTEOMICS TECHNOLOGY
Authors: Martin Matthew , Furamga Wieslaw , Mikhailov Valery
Published: June 2021

Background:

Lack of characteristic symptoms and adequate testing for pulmonary aspergillosis in immunocompromised host delays therapy, resulting in 80% mortality rate. Diagnosis requires biopsy with visualization of fungal hyphae and positive culture. It takes several days with 50% of false‐negative results. The non‐invasive assays (PCR, galactomannan, beta‐glucan) are impeded by false‐positive and false‐negative results. To find a better marker for pulmonary aspergillosis, we performed serum protein profiling on infected and non‐infected guinea pigs.

Methods:

Using SELDI‐TOF, serum was tested on CM10, Q10, H50, IMAC microchips and protein profile was obtained from Aspergillus infected and non‐infected animals terminated 1h and 3, 5, 7 days after inoculation. Difference in profiling was evaluated by hierarchical clustering and principal component analysis.

Results:

Two distinctive peaks in infected and one peak in non‐infected animals were identified. Future protein identification and validation as a potential diagnostic marker is warranted.

Conclusion:

Our study discovered exclusive spectrometric peaks that may contain potential biomarkers for early diagnosis of systemic aspergillosis.
3D molecular imaging of the “angiogenic switch” differentiates neovascular development in animal models of cancer
Authors: Caruthers Shelton D , Schmieder Anne H , Williams Todd A , Allen John S , Zhang Huiying , Winter Patrick M , Wickline Samuel A , Lanza Gregory M
Published: June 2021

The objective of this study was to determine whether neovascular progression in animal tumor models could be resolved and differentiated noninvasively with high‐resolution MR molecular imaging and 3D neovascular mapping techniques.

Methods

Rabbits implanted with Vx2 adenocarcinoma and nude mice implanted with MDA435 cells were imaged with MR at clinically relevant field strengths (3T) before and after administration with ανβ3−targeted paramagnetic nanoparticles. Tumor volumes and 3D MR neovascular maps were obtained in rabbits on days 8, 14 and 16 and in mice on days 14 and 21 post implantation.

Results

On day 8, angiogenesis comprised 0.9± 0.3% of the Vx2 tumor peripheral volume, which increased to 3.8±0.7% and 7.9±1.7% on days 14 and 16, respectively (p<0.05) (Fig). In mouse xenograft tumors, angiogenesis was minimal at 14 days (0.4±0.2%) and increased (p>0.05) to 1.0 ±0.5% at 21 days. In both models, nontargeted tumors had minimal enhancement.

Conclusion

These data suggest that tumor growth of the MDA‐435 tumor may be diminished due to a muted inflammatory response in comparison with the robust “angiogenic switch” observed in the Vx2 model. MR molecular imaging and 3D neovascular mapping could have an important contribution in personalized medical treatment regimens incorporating an antiangiogenic arm. Support: NIH/NCI U54 CA 119342
Validation of a hierarchical stem cell population in inflammatory breast cancer (IBC) by comparative genomic analysis
Authors: Barsky Sanford H. , Ye Yin , Little Sean , Sandy April , Xiao Yi
Published: June 2021

The existence of stem cells in human solid tumors has been presumed based on clinical observations and biomarker and clonal dilution studies. Our previous studies with a human xenograft model (MARY‐X) of IBC, a model that forms spheroids in vitro that resemble a blastocyst, have demonstrated the presence of a prominent stem cell‐like population of cells. Comparing MARY‐X with non‐IBC breast carcinoma and normal lines, we found known embryonal stem cell markers (Stellar, H19, Rex‐1, Nestin, CD133) present. RT‐PCR analysis also revealed the expression of OCT4, SOX2, and Nanog, transcriptional determinants essential for the pluripotency and self‐renewal of human embryonal stem cells. However these findings do not prove that there is truly a hierarchical reserve stem cell population since both stem cell markers and functions could be reversibly derived from the proliferating population. We reasoned that true hierarchical stem cells should have fewer genomic alterations (gains and losses) than the proliferating population. To this end we conducted array CGH and identified 10 regions of amplification and 10 regions of loss. We then derived probes from each of these regions and conducted multicolor FISH on individual cells disadhered from the spheroids. Each of the probes recognized a specific chromosomal region in normal HMECs. All of the probes derived from amplified regions or deleted regions showed marked amplification (5–20 fold) or loss in >99% of the cells. However there was a distinct subpopulation of <1% of human cancer cells that contained markedly fewer genomic alterations (gains or losses). The existence of this distinct subpopulation suggests that a truly hierarchical stem cell population exists in IBC. This comparative genomic approach may be generally applicable to the identification and study of stem cells in many solid tumors.
Site‐specific differential expression of endothelial KLF4 in athero‐susceptible and athero‐protective regions of porcine aorta
Authors: Fang Yun , Davies Peter F
Published: June 2021

The expression of Kruppel‐like factor 4 (KLF4) was investigated in athero‐susceptible and athero‐protective regions of swine aorta, locations that map to sites of disturbed (DF) or undisturbed (UF) flow profiles. Endothelial lysates were isolated from the inner curvature of the aortic arch (DF; athero‐susceptible) and a nearby UF region of the descending thoracic aorta (UF; athero‐protected). Endothelial KLF4 protein was expressed in cells of porcine aorta and vasa vasorum by immunohistochemistry. QRT‐PCR showed endothelial KLF4 transcripts to be up‐regulated 2‐fold in the atheroprotective region. Endothelial KLF4 protein in descending thoracic aorta was 35% higher than that in aortic arch by Western blot. Genome‐wide analysis of endothelial transcription profiles in athero‐susceptible and athero‐protective regions of swine aorta identified differential expression of the following putative downstream targets of KLF4: ‐ eNOS, MCP‐1, IL‐6, p53, p57/kip2, cyclin B1, and keratin 19, consistent with KLF4‐mediated transcriptional activation or suppression. To further elucidate the role of KLF4 in endothelial gene expression, KLF4 was overexpressed or suppressed in human aortic endothelial cells in vitro. The study implicates KLF4 as a contributory mechanism to regional athero‐protection. Supported by the AHA.
Aryl Hydrocarbon Receptor is Activated by High Glucose and Regulates the Thrombospondin‐1 Gene Promoter in Endothelial Cells
Authors: Dabir Pankaj , Marinic Tina , Krukovates Irene , Stenina Olga
Published: June 2021

Glucose levels correlate directly with the diabetic vascular complications, and the earliest sign of vascular complications is endothelial dysfunction. However, the transcriptional mechanisms activated by glucose in endothelial cells (EC) are poorly understood. Aryl Hydrocarbon Receptor (AhR), a receptor for xenobiotic compounds with unknown physiological function, was activated in EC by high glucose and, as shown by chromatin immunoprecipitation, bound the promoter of thrombospondin‐1, a potent anti‐angiogenic and pro‐atherogenic protein involved in development of diabetic vascular complications. A binding site for AhR was identified in the glucose‐responsive fragment of the promoter (−280/+66), and the deletion of 15 base pairs containing the binding site abrogated the response. The constitutively active rAhR induced activation of the TSP‐1 gene promoter more than 4 fold. In response to high glucose stimulation, AhR formed a complex with Egr‐1 and AP‐2, two other transcription factors activated by glucose in EC that have not been previously detected in complex with AhR. The activity of the DNA‐binding complex was regulated by glucose through the activation of the hexosamine pathway and intracellular glycosylation. This is the first report of activation of AhR by a physiological stimulus. This report links the activation of AhR to the pathological effects of hyperglycemia in the vasculature.
Soluble Epoxide Hydrolase Gene Deletion is Protective Against Myocardial Ischemia‐Reperfusion Injury In Vivo
Authors: Merkel Matthias Johannes , Iliff Jeffrey , Packwood William , Liu Lijuan , Cao Zhiping , Alkayed Nabil J , Van Winkle Donna M
Published: June 2021

Epoxyeicosatrienoic acids (EETs) are metabolized by soluble epoxide hydrolase (sEH). Targeted disruption of the sEH gene enhances post‐ischemic contractile function in isolated hearts and exogenous administration of EETs limits myocardial infarct size in vivo. We tested the hypothesis that sEH gene deletion is cardioprotective.

Male C57BL\6J wild‐type (WT) mice or sEH knockout (sEHKO) mice were subjected to 40 min coronary artery occlusion and 120 min reperfusion. Area‐at‐risk (AAR) and infarct size (I) were assessed using fluorescent microspheres and triphenyl tetrazolium chloride. Infarct size is expressed as I/AAR (mean ± SEM). Myocardial sEH protein content was assessed by immunoblot and its distribution by immunohistochemistry (IHC).

I/AAR was reduced by 25±9% in sEHKO compared to WT (37±3% n=10 vs. 49 ± 3% n=8, p<0.05). AAR/heart volume was comparable between groups (27 ± 4% vs. 29 ± 4%). Western blot confirmed the absence of sEH in sEHKO and its presence in WT hearts. IHC was positive for sEH in cardiomyocytes in WT but not sEHKO mice.

Cardioprotection from targeted deletion of sEH is likely due to increased endogenous myocardial EET levels. Infusion of EETs or inhibition of sEH may serve as a novel therapeutic option for cardioprotection during coronary revascularization and cardiac transplantation. Support: VA Merit Review 317 (DMVW) and RO1 NS44313 (NJA).
Proangiogenic properties of 4‐hydroxy phenylretinamide (4‐HPR) in experimental neovascularization and study of its mechanisms
Authors: Kannan Ram , Sreekumar Parameswaran G , Hinton David R
Published: June 2021

Purpose: We investigated the effect of 4‐HPR on a murine model of laser‐induced choroidal neovascularization (CNV). The effects of 4‐HPR on apoptosis and angiogenic factors was studied in cultured human retinal pigment epithelial cells (RPE).

Methods: Retina of C57BL/6 mice was exposed to laser irradiation. 4‐HPR (0.2 mg or 1 mg) or vehicle was injected intraperitoneally daily for 14 days post‐laser. Cultured RPE were treated with 4‐HPR and apoptosis and regulation of growth factors was determined.

Results: Mice treated with 4‐HPR exhibited an increase in vascular leakage, CNV lesion size, and lesion volume. A dose‐dependent decrease in lesion‐associated RPE cell number and increase in subretinal fluid was found. 4‐HPR increased RPE cytotoxicity, active caspase‐3 and apoptosis. Upregulation of vascular endothelial growth factor (VEGF)‐A, and ‐C and angiopoietin‐1 genes and increased VEGF secretion were observed with 4‐HPR (p<0.01 vs. untreated), while pigment epithelium derived growth factor and thrombospondin‐1 were significantly (p<0.05) downregulated.

Conclusions: Administration of 4‐HPR to mice resulted in increased CNV. The increased neovascularization with 4‐HPR was accompanied by upregulation of proangiogenic and downregulation of antiangiogenic factors. Loss of RPE around the laser lesions may arise from 4‐HPR‐induced cell death. Supported by Arnold and Mabel Beckman Foundation.
TSP‐1 Binds Thr92 Phosphorylated CD36 in a Calcium Dependent Manner and the Ligands of CD36 Regulate the Phosphorylation of Thr92 in Mouse Macrophages
Authors: Chu Ling‐yun , Silverstein Roy , Febbraio Maria
Published: June 2021

CD36 is an 88 kD transmembrane glycoprotein expressed on many cell types, including platelets, macrophages and endothelial cells. Our lab and others showed that CD36 is required for the anti‐angiogenic property of Thrombospondin‐1(TSP‐1), an extracellular matrix protein. Threonine 92 (Thr92) of CD36 is an important regulator of the binding of TSP‐1. Thr92 of CD36 is within a consensus protein kinase C (PKC) phosphorylation site and can be phosphorylated by PKC in vitro. The few published studies on CD36 phosphorylation in platelets suggest that the phosphorylated form, which has low affinity to TSP‐1, may be the default state. To study the phosphorylation of Thr92 of CD36, we cloned a 111bp CD36 fragment (FP111) that encodes amino acids 81 to 117. We showed that the WT but not T92A FP111 could be phosphorylated by PKC in vitro. FP111 binds TSP‐1 in vitro in a calcium dependent manner and the phosphorylation of FP111 enhances the binding of TSP‐1. Thr92 of CD36 in both elicited and resident mouse macrophages is phosphorylated, at least in part. Moreover, the phosphorylation level of CD36 in mouse macrophages can be regulated by CD36 ligands. Ectophosphorylation of CD36 is a novel mechanism of regulation and may impact CD36‐dependent functions. Determination of the effect and the regulation of CD36 phosphorylation will improve our understanding of the regulation of CD36 dependent functions, such as angiogenesis.