bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of a community‐based walking program on fruit and vegetable consumption
Authors: Hongu Nobuko , Block Linda M. , Sanchez Stephanie A. , Day Sharon Hoelscher , Harris Robin B.
Published: June 2021

The purpose of this study was to assess the impact of Walk Across Arizona, a 16 week, statewide outreach walking program, on self‐reported fruit and vegetable (F&V) consumption in adult participants (18–100 yrs). F&V consumption was assessed in pre‐ and post‐program self‐administered surveys that asked amount and what kinds of F&V eaten in the previous day. Changes in F&V consumption were assessed using Wilcoxon matched‐pairs signed‐rank tests. Potential impact of age, body mass index (BMI), and prior physical activity were assessed using multivariate regression. Of the 2,454 adult participants registered for the program, 1,170 (70% female, mean age 50.0 years) reported F&V consumption at both registration and wrap‐up surveys. These participants significantly increased their reported consumption of F&V (p<0.05), while increasing their physical activity level and decreasing their BMI. The change in F&V consumption during the program was associated with baseline F&V consumption, gender, ethnicity, and the average miles reported per week. These findings suggest that a program aimed at increasing physical activity can have an impact on other areas of healthy living in the form of increased F&V consumption. The integration of nutrition messages into the walking program may further increase the consumption of F&V amongst program participants.
Oral iron supplementation with a micronutrient powder (Sprinkles®) provided to children with moderate‐to‐severe malnutrition does not increase infectious morbidity risk: a randomized, double‐blind, placebo‐controlled, non‐inferiority safety trial
Authors: Lemaire Mathieu , Abedin Fahmida , Shen Hua , Islam Qazi Shafayetul , Aziz Marufa , Haseen Farhana , Parveen Monira , Chowdhury Jalal , Hyder Ziauddin , Cook Richard J , Zlotkin Stanley H
Published: June 2021

Background

A link between iron supplementation (IS) and infectious morbidity (IM) has been suggested, particularly in children with moderate to severe malnutrition (MSM). Two meta‐analyses concluded that oral IS is safe, but children with MSM were excluded.

Methods

This is a randomized, double‐blind, placebo‐controlled, non‐inferiority (NI) safety trial using a 2‐mo course of iron‐containing micronutrient or placebo powder (I‐MNP and PP) in 268 Bangladeshi children aged 12–24 mo with MSM (weight‐forage Z‐score ≤ −2). NI margin was set at 20%. Primary outcome was a composite of diarrhea, dysentery and lower respiratory tract infection episodes (D/D/L) recorded through home visits every 2 days (first 2 mo) and then twice a week (next 4 mo post‐intervention). To capture seasonal effects, one group was studied in the summer and another in the winter. Univariate Anderson‐Gill model was used to analyze recurrent events.

Results

Baseline characteristics were similar. Combined relative risk (RR) of D/D/L for I‐MNP vs. PP was 0.81 (95% confidence interval [CI] 0.62–1.04; p=0.09). Winter group RR was lower at 0.6 (CI 0.42–0.85; p=0.004) than summer group RR, at 0.98 (CI 0.69–1.38, p=0.9), p=0.052. There were no reported deaths.

Conclusion

Risk of IM was not higher for I‐MNP vs. PP. Since IM rates were lower than anticipated, NI is not confirmed. I‐MNP was more protective for the winter group, suggesting a seasonal effect.

Grant Funding Source: Thrasher Research Fund
A Pilot Study of Paraprofessional Peer Teaching by Teen Moms in A Teen Parenting Nutrition Education Program
Authors: Wong Siew Sun , Keller Sylvia D
Published: June 2021

Objective

Measure the effectiveness of a peer‐teaching‐peer paraprofessional model (PTPPM) in a teen parenting nutrition program.

Methods

Two teen moms were trained as paraprofessional peer educators (PPE) for The CARE42 program in two sites. Site 1 consisted of predominantly non‐Hispanic White females whereas Site 2, predominantly Hispanic females. Each PPE taught 10–13 participants (aged 14 to 19, single, non‐pregnant females) per site over three weeks. Each lesson was 30 minutes. An RD attended every lesson, evaluated teaching, and facilitated in Q&A. For evaluation among PPE and participants, the study used pre/post tests to assess improvements in nutrition knowledge, diet, physical activity, behavior, and self‐esteem (NK‐D‐PA‐SE).

Results

PPE improved significantly in diet, physical activity, behavior, and self‐esteem. Although not significantly different, PPE's teaching accuracy score was high and improved consistently. Among 28 recruited participants, 23 completed the study. Both sites had significant improvement in NK‐D‐PA‐SE. These positive changes indicated that PPE were able to teach their peers effectively. 83% of participants liked or very much liked to be taught by their peers.

Conclusions

Teen moms aged 18–22 are capable of teaching their peers accurate nutrition information. This PTPPM has great potential in bringing sound nutrition education to teen parents in a motivating way.
Teachers perceive “California Nutrition Education Competencies, Pre‐Kindergarten through Grade Twelve” as a useful framework for classroom instruction
Authors: Briggs Marilyn M. , Reed Heather , Zidenberg‐Cherr Sheri
Published: June 2021

This study assessed teachers’ perceptions related to the revised “California Nutrition Education Competencies, Pre‐Kindergarten through Grade 12” (NEC). The NEC provides nutrition content standards for classroom instruction and is aligned with California core academic content standards and the 2009 Health Education Content Standards for California Public Schools” (HECS). A random sample of California public school teachers (n=156) were surveyed regarding perceptions and practices related to the NEC. The majority of teachers responded that the NEC was organized in a sequential manner (93.5%), well structured (82.5%), grade‐level appropriate (85.7%–97.7%), consistent with the grade‐level expectations of the HECS (81.2%), useful for selecting appropriate nutrition topics for their grade level (86.4%), and valuable as a resource for teachers (90.9%). In conclusion, teachers perceive the NEC as a useful and appropriate framework for incorporating nutrition education into classroom instruction. Next steps will include research to determine effective methods to increase the use of the NEC in providing nutrition‐related instruction in California classrooms. Funding for this study was provided by the California Department of Education, Nutrition Services Division.
Garden‐enhanced nutrition education improves nutrition knowledge and behavior in second‐grade students
Authors: Jones Anna , Ly Carol , Wasilewski Lisa , Zidenberg‐Cherr Sheri
Published: June 2021

This study assessed the impact of the Healthy Eating Active Living (HEAL) nutrition curriculum on improving the nutrition and health knowledge and vegetable preferences of second grade children. Second graders and their parents in three schools in San Mateo County, California, participated in the study. Children received 21 weekly garden‐enhanced nutrition education lessons and participated in active games twice weekly during lunchtime. Knowledge and behavior surveys were administered to children pre‐ and post‐intervention, and a behavior survey was completed by parents, pre‐ and post‐intervention. Mean nutrition knowledge increased among students (p<0.001). Out of the five vegetables tasted by students, only zucchini preference increased (p=0.016). Parents reported their child increased consumption of other fruits (p=0.006), and decreased consumption of cookies (p=0.000). They also reported an increase in their child's enjoyment of active games after intervention (p=0.001). In summary, 21 weeks of exposure to the HEAL curriculum increased nutrition knowledge, improved preference for zucchini, increased consumption of fruit, and decreased consumption of cookies. The HEAL curriculum is an effective way to improve knowledge and behavior and is popular with students. Funded in part by the San Mateo County Department of Public Health.
Identification of Food Sources Contributing to Dietary Quality in the Lower Mississippi Delta Region: A Systematic Approach to Designing Dietary Modifications for Obesity Prevention
Authors: Onufrak Stephen John , Connell Carol , Zoellner Jamie , Thomson Jessica , Champagne Catherine , Bogle Margaret L , Yadrick M Kathy
Published: June 2021

The Lower Mississippi Delta (LMD) region suffers from high rates of obesity and chronic diseases.

Our objective was to design culturally appropriate dietary modifications for the prevention of obesity in the LMD population based upon the 2005 Dietary Guidelines for Americans (DGA) and observed eating patterns.

Using data from the Foods of our Delta 2000 study, we evaluated adult dietary quality using the 2005 Healthy Eating Index (HEI‐2005), determined leading food sources contributing to HEI‐2005 components relevant to obesity, and investigated demographic differences in food source intakes. This information was used to construct specific dietary modifications to improve adherence to the DGA within the LMD population.

Total HEI‐2005 score for the LMD excluding sodium score was lower than the 1999–2000 NHANES population (47.4 versus 51.7, p=0.0006) with lower scores for whole fruit, total vegetables, dark green/orange vegetables and legumes, and milk. Younger age was the largest determinant of poor dietary quality in the LMD with total score and 8 component scores decreasing with decreasing age; 20 of 35 food sources also differed by age. Targeted dietary modifications to address observed DGA inadequacies were constructed based upon commonly consumed foods.

A systematic approach based upon observed intake is useful in the construction of culturally appropriate dietary modifications for a given population.
Individually tailored nutrition program is effective in improvement of chronic disease risk factors in worksite settings
Authors: Park Handeuk , Kim EunJin , Hwang Myungok , Paek Yun‐mi , Choi Tae‐in , Park Yoo Kyoung
Published: June 2021

The incidence of chronic disease is continuously increasing in Korea. Especially, office workers have higher risk of chronic disease because of their dietary habit and lifestyle. The study aimed to investigate the effect of tailored nutrition counseling on improving chronic disease risk factors. Ninety‐nine male workers (age 46.9 ± 7.0yrs) volunteered for 12 weeks of nutrition program containing dietary intake and physical activity adjustment. Five individualized program was performed with the main theme of wt loss (OB, n=16), blood pressure lowering (BP, n=34), normalizing blood glucose (FPG, n=21), lipid lowering (LPD, n=13) and reducing MS risk factors (MS, n=15). Anthropometric data, blood‐pressure, self‐reported questionnaire, blood profiles were measured before and after 12weeks of nutrition education. The education program included 5 times of 1:1 interview. Compared to 0 week, anthropometric data (weight, BMI, fat, visceral fat, waist, SBP, DBP) were significantly decreased after 12 weeks (p<0.001). Blood fasting blood glucose, total cholesterol were decreased (p<0.05). The primary outcomes with individually tailored programs showed to be more effective than one general nutrition program.
Associations between bone quality and restrained eating, body weight, diet and physical activity in young adult Caucasian females
Authors: Zanovec Michael , Tuuri Georgianna
Published: June 2021

This study examined the associations between bone quality estimated from quantitative ultrasound (QUS) and self‐reported diet, eating behaviors, physical activity (PA), and lifestyle factors. Fifty‐eight Caucasian females 19–24 years of age participated. Subjects completed three scans of the right heel using a GE Lunar Achilles InSight QUS machine; a semi‐quantitative modified Block food frequency questionnaire (FFQ) with approximately 105 items; ten questions regarding restrained eating (RE); the International Physical Activity Questionnaire (IPAQ) short form; a health history form; and were measured for height and weight. Factor analysis was used to determine relationships between variables. QUS scores factored with RE and weight; height was related to fiber; and neither total energy (kcal) nor PA were related to QUS or RE. Further research is needed with more sensitive bone markers and a larger sample size to determine the relationship of bone quality with diet, PA, and eating behaviors in young adult females.

Funded in part by Federal Hatch Act Funds #LAB93846.
Higher protein weight loss diet does not affect bone mineral density (BMD) in a double‐blind RCT in postmenopausal women
Authors: Thorpe Matthew P , Mojtahedi Mina C , Richey Abby L , Layman Donald K , Evans Ellen M
Published: June 2021

In spite of longstanding theory that protein intakes above the RDA are harmful to bone, clinical trials have variably shown no influence or protection of bone with higher protein intakes during weight loss. Studies to date have failed to match intakes of calcium and vitamin D, leaving the independent effect of protein ambiguous.

We prescribed a conventional, MyPyramid‐based energy restricted diet (1500 kcal/d) to 31 postmenopausal women (age 65±5y, BMI 33.8±4.9), then randomly assigned 50g/d of supplemental maltodextrin (CARB) or whey protein isolate (PRO) to be added as 25g doses to breakfast and lunch, for 0.8g/kg total protein in CARB and 1.4g/kg in PRO groups. All women received 1g Calcium and 800 IU Vitamin D as a daily multivitamin.

After 6 mo, women lost (PRO, CARB) 9.8±6.7, 4.8±3.7% of baseline weight (p=0.06). Bone mineral density changed 2.4, 2.1% (p=0.70) for whole body, −0.8, 1.4% (p=0.13) for lumbar spine and −0.4 and −0.2% (p=0.84) for total hip.

This double blind, randomized, controlled trial supports growing consensus that higher protein intakes are not harmful to bone in the context of adequate calcium and vitamin D intakes.

Support: National Dairy Council & Carraway Foundation
Anti‐oxidation activities of CordyMax in an oxidative stress model: A mechanism of its anti‐aging property
Authors: Yang Jieying , Tan Ningzhi , Zhao Chunsheng , Zhang Yan , Zhu Jia‐Shi
Published: June 2021

Cordyceps sinensis (Cs) is traditionally believed as an anti‐aging herb. CordyMax (CM), a mycelia fermentation product of Cs, has shown the benefits of lifespan extension, anti‐fatigue and improving glucose‐lipid‐energy metabolisms. We further tested the antioxidant function of CM in mice, the mechanism of its anti‐aging effect. Mice were randomized into 5 groups, receiving vehicle or CM at a dose of 0.5, 1.0 or 1.5 g/kg for 60 days by gavage. They were given a single dose of 11 Gry 60Co γ‐radiation on Day 60 and sacrificed on Day 64. Compared to non‐radiation controls, γ‐radiation reduced plasma glutathione (GSH) & the thiol groups, liver catalase (CAT) & SOD, GSH‐peroxidase (GSH‐Px), GSH‐reductase (GSH‐Rd), and increased liver protein carbonyl groups. Compared to radiation controls, 60 days of oral CM increased plasma thiol groups by 21–25% in the 3 dose groups, and liver CAT by 15–16% (all p<0.01). Oral CM at a dose of 0.5 or 1.0g/kg increased GSH‐Px by 15% (p=0.01). CM therapy (1.0 or 1.5g/kg) reduced liver protein carbonyl groups by 9–14% (p=0.04, <0.01) and 8‐OHdG by 26–34% (p<0.01); and increased plasma GSH by 26–27% (p=0.02, 0.03), liver GSH‐Rd by 10–11% (p=0.02) and liver SOD by 6–9% (p<0.05). In conclusion, oral CM improves antioxidant capacity in mice with radiation‐induced oxidative injure, supporting in general the anti‐aging activity of CM.
A randomized study of legume consumption during weight loss: effects on food cravings
Authors: McCrory Megan A , Lovejoy Jennifer C , Palmer Philip A , Eichelsdoerfer Petra E , Gehrke Malinda M , Kavanaugh Ian T
Published: June 2021

To examine the associations of weight loss and dietary composition with food cravings, we studied n=42 adults (BMI 25–35 kg/m2) who completed a 6‐wk study. Subjects were assigned to consume either LOW (1 Tbsp), medium (MED, 0.5 c), or HIGH legumes (1.8 c (women) and 2.7 c (men)) diet 6 d/wk while reducing energy intake (EI) by 30%. ~50% of EI was provided and contained the requisite amount of legumes, while the remainder of EI was self‐selected. All groups lost weight (p=0.023), with MED losing more than LOW (p=0.032) but not HIGH (p=0.12) (meanΔ±SEM: MED: −3.8±0.5 kg (n=16); HIGH: −2.3±0.6 kg (n=13); LOW: 1.8±0.5 kg (n=13). At baseline, 76% of subjects had food cravings (Hill et al questionnaire, 1991) over the past 3 months, averaging 11.1±1.7 times/mo. The change in the frequency of the strongest food craving differed among groups, decreasing by 5.3±3.9 and 0.7±1.0/mo in LOW and MED, respectively, and increasing by 2.5±2.5/mo in HIGH (p=0.069). The BMI decrease was significantly associated with the decrease in the frequency of the strongest food craving after controlling for legume group, baseline BMI and baseline strongest craving frequency the (p=0.049). All groups consumed the most craved food significantly less often at wk 6 compared to wk 0 (Δ= −15±7% of the time; p=0.013), but acting on cravings was not related to weight loss. Food craving frequency can change with weight loss and changes in dietary composition. [Funding: Pulse Canada PIP]
Human biospecimens collection in the era of Personalized Medicine: challenges and perspectives
Authors: Potapova Olga
Published: June 2021

Why do we need your tissues? Modern biomedical research requires use of clinically characterized, high‐quality human biospecimens yielding high‐integrity analytes for development of novel medicines and companion diagnostic tests, biomarker assay development and, overall, achieving the goals of the personalized medicine. Effects of most important pre‐analytical biospecimen variables, significantly confounding human tissue research will be discussed: tissue collection specifications, processing materials and methods, storage and shipping procedures, molecular analyte preparation techniques, etc. Other issues include: collection of biospecimens using standard protocols versus project‐based collection protocols; using currently available biorepositories for future studies on the indicated subject; relevant regulatory and legal issues, including international disparities in regulations on using human materials for biomedical research.
Effect of Tamoxifen and Raloxifene on platelet activation markers
Authors: Dobrydneva Yuliya , Vishneski Susan R.
Published: June 2021

Tamoxifen (TAM) and Raloxifene (RAL) are structurally related non‐steroidal estrogens used for prevention of breast cancer. However, thrombosis is as an adverse effect of both drugs. The goal of the study was to determine effect of TAM and RAL on platelets in whole unfractionated blood. Activation markers expression (Annexin V binding, P‐selectin expression and PAC‐1 binding) in TAM‐ and RAL‐ treated platelets was measured. 1uM TAM increased double positive platelets (CD62/PAC‐1) by 19±5% while 1uM RAL increased by 20±7%. In combination with 1uM TRAP6, both TAM and RAL at 10 uM increased TRAP6 induced elevation of CD62/PAC‐1 expression (TRAP6, 43±11%; TAM+TRAP6, 108±44%; RAL+TRAP6, 120±39%). Both TAM and RAL increased Annexin V binding (1.3±0.6% baseline, 10 uM TAM 4.7±1.5%, 10 uM RAL 5.2±1.7%). ADP at 5 uM induced 2.7±0.7% Annexin V binding; ADP+10 uM TAM was 5.8±1.7%, and ADP+10 uM RAL was 7±2%. Both drugs stimulate expression of platelet activation markers and sensitize platelets to agonists. This provides a possible mechanism of thrombosis in patients taking RAL and TAM for the prevention of breast cancer (Supported by the SDG from the AHA, CHRB and Cancer Research and Prevention Foundation).
Biochip Array Analysis of Various Mediators of Inflammation in Disseminated Intravascular Coagulation
Authors: Rahman Saud , Hoppensteadt Debra , Cunanan Josephine , Davis Rachael , Sadeghi Nasir , Fareed Jawed
Published: June 2021

Disseminated intravascular coagulation is a polypathologic syndrome which involves blood, endothelial and target organ dysfunction resulting in the generation of various mediators of vascular dysfunction, hemostatic aberrations and hemodynamic disturbances. In addition, various disorders of target organs such as kidney, liver, heart and brain are observed. Uncontrolled protease generation results in the formation of various mediators of inflammation such as neuron specific enolase (NSE), neutrophil gelatinase associated lipocalin (NGal) and soluble tumor necrosis factor receptor 1 (TNF R1). Endothelial damage results in the generation of thrombomodulin (TM) and endogenous coagulation/fibrinolysis results in D‐ Dimer formation. In order to study the circulating levels of these mediators, a Biochip array method (Randox, Oceanside, CA) was utilized with samples obtained from clinically diagnosed DIC (n=100) and normal individuals (n=10). Circulating levels of these mediators were markedly increased in DIC patients in comparison to normals. The most striking increase was noted in NGal (4–6 fold) and TNF R1 (10 fold). Other mediators were also increased in the DIC group, however the data was broadly scattered. These results indicate that NGal and TNF R1 along with other mediators play a major role in the pathophysiology of DIC.
Comparative Studies on the USP Reference Standard Over‐Sulfated Chondroitin Sulfate and Two Semi‐synthetic Over‐Sulfated Chondroitin Sulfate Preparations
Authors: Sadeghi Nasir , Hoppensteadt Debra , Iqbal Omer , Jeske Walter , Walenga Jeanine M. , Fareed Jawed
Published: June 2021

The reported heparin contaminant was identified as OSCS. Because of structural differences in OSCS preparations, the USP has introduced a reference standard. The purpose of this study was to compare the USP standard with commercially available OSCS and a semi‐synthetic OSCS. Unfractionated heparin (UFH) was used as a reference preparation. The three OSCS preparations were profiled for molecular weight using HPLC. The anticoagulant activities were measured using whole blood activated clotting time (ACT) and plasma based assays (APTT, Heptest, PT, thrombin time and amidolytic anti‐IIa). All OSCS preparations showed a comparable molecular profile with molecular weights ranging from 19.8 kDa to 20.8 kDa. Heparin showed a slightly lower molecular weight (17.2 kDa). In the whole blood ACT, USP reference showed the stronger anticoagulant activity compared to Serva and semi‐synthetic OSCS. In the PT and Heptest no differences were noted in three agents, however in the aPPT and TT assay, USP showed markedly stronger activity than other two products. In the amidolytic anti‐IIa activity, IC50 value ranged from 8.5 to 14.5 μg/ml. These results showed that although all of the OSCS had a comparable molecular profile, the anticoagulant activity of these preparations varied widely. USP reference standard OSCS showed the strongest activity. Therefore, the other biologic profile of the USP reference OSCS may also be different.
Comparative Studies on Branded Dalteparin with Two Generic Versions: Hepagumin and Fluzepamin
Authors: Sadeghi Nasir , Hoppensteadt Debra , Iqbal Omer , Jeske Walter , Walenga Jeanine M. , Fareed Jawed
Published: June 2021

Dalteparin is a low molecular weight heparin (LMWH) prepared by nitrous acid digestion. Recently generic versions of the product have become available. The purpose of this study was to compare the molecular and functional profile of Dalteparin and two generic versions from Japan (Hepagumin and Fluzepamin). The molecular profile was determined using the HPLC before and after heparinase‐1 digestion. The anticoagulant activity was measured using the whole blood ACT‐Hemochron and plasma‐based APTT and Heptest, and amidolytic anti‐Xa and anti‐IIa assays. Thrombin generation studies were performed using a kinetic method. By HPLC, the molecular weight of the three products was comparable and ranged from 6.3 to 6.8 kDa. After heparinase digestion all three products give comparable result (molecular weight ranging from 2.1 to 2.3 kDa). The oligosaccharide components after digestion were also comparable. In the whole blood anticoagulant studies no differences were noted between products at the 1.25 U/ml. In the APPT and anti‐Xa assay, there were no differences between three products. However, minor differences were noted in the Heptest and factor IIa inhibition. These results show that Hepagumin and Fluzepamin do not significantly differ from the branded dalteparin. The minor differences noted in clot based assays were not statistically different.
Potency Adjustment of Unfractionated Heparins to Harmonize the USP and WHO Standardization. Impact on Anticoagulant Practice in the US
Authors: Hoppensteadt Debra , Sadeghi Nasir , Iqbal Omer , Wahi Rakesh , Lewis Bruce , Fareed Jawed
Published: June 2021

To harmonize the potency of US heparin (UFH) supplies with the WHO standard, the USP has mandated use of a revised procedure resulting in a 10–13% reduction of anticoagulant potency. To assess the impact of 10% potency adjustment, stock solutions of UFH were prepared to represent a 10% decrease (90 U/ml), no change (100 U/ml) and a 10% increase (110 U/ml). Differences were evaluated in the surgical and interventional range (1–5 U/ml) using the ACT and in the therapeutic range (0–1.0 U/ml) using plasma clotting assays. Protamine titration studies were conducted in both studies. Thrombin generation studies were conducted at lower levels. Rabbit models of stasis thrombosis and ear bleeding were used to determine the impact on antithrombotic and hemorrhagic activities. Differences in the ACT were only noted at concentrations >1.0 U/ml. No difference in protamine neutralization profile was noted. Supplementation to normal plasma did not result in differences in the PT, APTT, Heptest, thrombin time, anti‐IIa and anti‐Xa activities. Some differences were noted utilizing a thrombin generation assay. These studies suggest that a 10% change in heparin potency may not impact the anticoagulant practice in the therapeutic or prophylactic range. Closer monitoring may be needed in specialized populations and in those patients requiring surgical and prophylactic anticoagulation where higher concentrations of UFH are reached.
Post‐secretory activation of Paneth cell α‐defensins in the cecal and colonic lumen of matrix metalloproteinase‐7‐null mice
Authors: Mastroianni Jennifer Rose , Costales Jessica K. , Salzman Nita H. , Ouellette Andre J.
Published: June 2021

In mice, Paneth cell α‐defensins, termed cryptdins (Crps), are converted intracellularly from inactive precursors to bactericidal forms by matrix metalloproteinase‐7 (MMP‐7). Because it is possible that other extracellular proteinases may be capable of similar processing, we tested whether secreted proCrps are converted to active forms in the MMP‐7−/− mouse. Protein extracts from complete (organ plus luminal contents) ileum, cecum, and colon of MMP‐7−/− and wild‐type C57Bl/6 mice were compared by gel permeation chromatography and acid‐urea (AU)‐PAGE. As anticipated, MMP‐7−/− ileum lacked appreciable levels of processed Crps. However, high levels of apparent Crps were purified from the cecum and colon of MMP‐7−/− mice. N‐terminal sequencing and mass spectrometry identified α‐defensins consisting of a mixture of native Crps, N‐terminally truncated Crps, and Crps with elongated N termini that retain proregion residues near the proregion‐Crp junction. Thus, secreted mouse proCrps may be processed in the large bowel lumen, even in the absence of the intracellular activating enzyme. Also, the MMP‐7−/− mouse may be a useful model of innate mucosal immunity in the small intestine, but the knockout is not α‐defensin deficient in cecum or distal colon, downstream from the site of precursor secretion in small intestinal crypts of Lieberkühn.

Supported by NIH grants DK044632, AI059346, and AI057757.
Hyaluronan‐chondrocyte interactions mediate cell signaling pathways
Authors: Luo Na , Knudson Cheryl B , Knudson Warren
Published: June 2021

Osteoarthritis is a degenerative process of articular cartilage. The focus of our project is to identify strategies to stimulate cartilage matrix repair. An intact, hyaluronan‐dependent pericellular matrix promotes the chondrocyte response to BMP‐7. Disruption of chondrocyte‐matrix interactions occurs during disease progression. The purpose of this study is to validate the function of CD44, the principal hyaluronan receptor, in mediating the effects of hyaluronan on chondrocyte signaling. Primary chondrocytes were isolated from 6–8 day old BALB/c mice and CD44 knock‐out (KO) mice, as well as adult bovine articular cartilage (CD44 positive). Testicular hyaluronidase (T H'ase) treatment of chondrocytes removed pericellular hyaluronan as visualized by immunostaining. Pre‐treatment with T H'ase stimulated Akt & ERK phosphorylation; this stimulation was muted by CD44 siRNA treatment of bovine chondrocytes. However, pre‐treatment with T H'ase reduced Smad1 phosphorylation and Id2 protein stimulation in response to BMP‐7 in all three chondrocyte models. Thus, in mouse chondrocytes, removal of hyaluronan has a profound effect on cell signaling pathways regardless of endogenous CD44 expression. Receptor compensation may occur in the CD44 KO chondrocytes or alternative mechanisms of hyaluronan‐cell interaction may influence these signaling pathways. Supported by NIH R01‐AR39507 and R01‐AR43384.
CHARACTERIZATION OF THE OPIOID PEPTIDE DYNORPHIN A AS A NOVEL LIGAND FOR LEUKOCYTE INTEGRIN αMβ2 (Mac‐1)
Authors: Brothwell Julie , Podolnikova Nataly P. , Ugarova Tatiana P.
Published: June 2021

Substantial evidence indicates that opioid peptides, including dynorphin A (Dyn A 1–17), induce leukocyte migration, their degranulation and cytokine production. The mechanisms whereby opioid peptides induce leukocyte responses are unknown. The integrin αMβ2 (Mac‐1, CD11b/CD18) is a multiligand receptor which mediates numerous reactions of leukocytes during the immune‐inflammatory response. Our recent elucidation of αMβ2 recognition specificity suggested that Dyn A contains the αMβ2 recognition motifs and can potentially interact with the receptor. In this study, we synthesized peptide libraries spanning the sequence of big dynorphin (1–31) and demonstrated that several dynorphin‐derived peptides bound recombinant 125I‐labeled αMI‐domain, the ligand binding domain of αMβ2. We have further shown that synthetic peptide duplicating Dyn A supported adhesion of the αMβ2‐expressing cells and soluble peptide inhibited adhesion. In binding to Dyn A, αMβ2 cooperated with cell surface proteoglycans since anti‐αMβ2 specific antibodies and glycosaminoglycans were required to block adhesion. The interaction of Dyn A with the αMI‐domain was activation‐independent as both the α7 helix‐truncated and helix‐extended I‐domains efficiently bound Dyn A. The results identify dynorphin as a novel ligand for αMβ2 and suggest that the interaction of αMβ2 with opioid peptides may be involved in the neuroimmune axis.
The Potential of Conditionally Replicating Adenovirus Expressing TIMP2 for Ovarian Cancer Therapy
Authors: Yang Sherry , Douglas Joanne T. , Alvarez Ronald D , Siegal Gene P , Ponnazhagan Selvarangan
Published: June 2021

Current therapies for ovarian cancer have limited prognostic outcome due to advanced stage of the disease at diagnosis. Among new therapies, conditionally replicating adenoviruses (CRAds), designed to selectively lyse cancer cells, hold promise. However in clinical trials, CRAds exhibit limited efficacy thus far. Second generation CRAds are being developed to express a therapeutic protein to enhance antitumor efficacy. One attractive target in the tumor microenvironment is the matrix metalloproteinases (MMPs). MMPs are endogenous proteases that degrade ECM and are upregulated in ovarian cancer. Tissue inhibitors of metalloproteinase 2 (TIMP2) is an endogenous inhibitor of MMPs. The present study developed a CRAd that uses the CXCR4 promoter for enhanced replication combined with the inclusion of the TIMP2 transgene. Transductional selectivity is further achieved by replacement of the Ad5 knob with the Ad3 knob, whose receptor is overexpressed on ovarian cancer cells. To evaluate the expression profile of this armed CRAd, human ovarian cancer cells were infected with Ad5/3‐CXCR4‐TIMP2. The TIMP2 produced was functional, as demonstrated by the inhibition of MMP activity. Further validation of this virus on primary ovarian cancer tissues obtained from patients revealed high levels of viral replication, suggesting the potential of this vector for ovarian cancer therapy.
Bowel morphologic evaluation in a model of experimental intoxication with 1‐naphtyl‐N‐methylcarbamate
Authors: Cavalcanti Karen Pena , Souza Ivone Antonia , Castro Ricardo , Teles Nicodemos , Cabrita Antonio Silverio
Published: June 2021

The 1‐naphtyl N‐methylcarbamate (1NC) is one general purpose insecticide, with a wide specter of action. It is considered to have low toxicity therefore. The present work evaluates the bowel lesions in experimental administration of 1NC.

Twenty Wistar males rats 8 weeks of age were distributed randomly in 2 groups with 10 animals each. Group I constituted a control group with no manipulation. Group II constituted a group submitted to the administration of 1NC in drinking water in concentration of 60mg/l during 8 weeks. All the animals were sacrificed and by the end of twenty‐eight weeks. The bowel of control had no lesions. The macroscopic evaluation of the bowel showed aberrant crypts (AC) and neoplasms (N), in group II. The incidence of AC observed in this group was 6.27 ± 4.92 in the rectum, 16.87 ± 10.27 in the colon and 11.3 ± 8.49 in the cecum. The distribution of AC in this group was: 10,4% ± 9,24 in the rectum, 73.78% ± 8.01 in colon and 15,83% ± 8.01 in the cecum. The distribution of the neoplasm was of 80% in the colon and 20% in the cecum, 8 were observed in the colon and 2 in the cecum. By microscopy all the neoplastic lesions were adenocarcinomas.
Adventitial Infiltration of Activated Macrophages (MΦ) in Mesenteric Arteries of DOCA‐salt Rats
Authors: Thang Loc Vinh , Galligan James J
Published: June 2021

Sympathetic nerve activity is increased in DOCA‐salt hypertension. Furthermore, local mechanisms regulating neurotransmitter release from sympathetic nerves supplying mesenteric arteries (MA) are impaired in DOCA‐salt hypertension. DOCA‐salt hypertension is associated with oxidative stress and elevated vascular levels of superoxide (O2−). In mice deficient in Mφ colony‐stimulating factor, exhibit reduced inflammation and has vascular protection against damage caused by DOCA‐salt hypertension. We tested the hypothesis that there is an increase in Mφ infiltration into the adventitia of the MA of DOCA‐salt rats and this contributes to the increased O2− that might disrupt sympathetic nerve function. We found an increased number of Mφ in the peritoneal cavity and in the adventitia of MA in DOCA‐salt rats compared to sham rats (p<0.05). Many Mφ were localized close to sympathetic nerve fibers. In addition, the perivascular Mφ of DOCA‐salt rats had a higher mean fluorescence intensity of p22Phox, a subunit of the O2− generating enzyme NADPH oxidase, and TNF‐α compared to sham rats (p<0.05). Therefore, suggesting an increase in activated Mφ in the adventitia of MA from DOCA‐salt rats. The close proximity of Mφ to sympathetic nerve fibers suggests that the inflammatory cytokine TNF‐α and O2− released by Mφ could disrupt sympathetic neurotransmission in MA in DOCA‐salt rats. (Supported by HL70687)
Mitogen‐Activated Protein Kinase Phosphatase‐1 (MKP‐1) Is Low In Dorsal Medulla Of Hypertensive (mRen2)27 Transgenic Rats
Authors: Nautiyal Manisha , Tallant E. Ann , Chappell Mark C , Diz Debra I
Published: June 2021

Angiotensin II (Ang II) activates mitogen‐activated protein kinase (MAPK) pathways, whereas emerging evidence suggests that Ang‐(1‐7) up‐regulates MAPK phosphatases to reduce MAPK activities. Imbalances in Ang II and Ang‐(1‐7) favoring Ang II are implicated in (mRen2)27 [mRen] hypertensive transgenic rats which contain low medullary Ang‐(1‐7); replacement of Ang‐(1‐7) lowers pressure, improves reflex function and increases MKP‐1 mRNA in the dorsal medulla. Hypotensive ASrAOGEN (AS) transgenic rats have enhanced reflex function and high Ang‐(1‐7). We compared expression and activity of MKP‐1 in dorsal medullary tissue of mRen and AS rats. Western blot analysis shows that MKP‐1 is lower in mRen compared to AS [mRen: 0.69 ± 0.11 vs AS: 1.09 ± 0.05; n = 3, p = 0.03]. In contrast, auto‐phosphorylated, activated extracellular‐signal‐regulated kinase (ERK1/2), a substrate for MKP‐1, tended to be higher in mRen vs AS medulla. The ratio of MKP‐1 to ERK1/2 is 0.24 ± 0.02 in mRen and 0.92 ± 0.24 in AS rats (p = 0.04). Thus, signaling in hypertensive mRen is shifted towards the kinases, whereas MKP‐1 is higher in AS that have lower resting systolic pressure and enhanced reflex function. These results suggest that Ang‐(1‐7) up‐regulates MKP‐1 to counter‐balance Ang II‐stimulated MAPK activity and improve cardiovascular function.
Rapid estrogen signaling via GPER in human coronary artery smooth muscle
Authors: Han Guichun , Ma Handong , Barman Scott A. , Sellers Minga , Yu Xuan , Stallone John N. , White Richard E.
Published: June 2021

Estrogen produces rapid, non‐genomic effects in coronary arteries, but the mechanisms underlying these responses are poorly understood. We had previously demonstrated that rapid vasodilatory effects of 17 beta‐estradiol on coronary arteries were mediated via the PI3 kinase – Akt signaling cascade (Han et al., Am. J. Physiol. 293:H314, 2007). We now identify a mechanism of acute estrogen signaling in human coronary artery smooth muscle cells (HCASMC) involving the G protein‐coupled estrogen receptor (GPER). HCASMC were grown in short‐term culture, and the effects of the GPER agonist, G1, were measured on activation of important downstream signaling molecules. Immunoblot studies demonstrated both basal and G1‐stimulated activation of mitogen‐activated protein kinase in HCASMC, and revealed that G1 acutely (2–5 minutes) stimulated phosphorylation of ERK1/2. Further, our studies also revealed that G1 stimulates Akt phosphorylation in HCASMC. We also found that G1, like 17beta‐estradiol, relaxed porcine arteries in vitro. These studies indicate a novel signaling mechanism for acute estrogen effects in human coronary arteries: stimulation of GPER, with subsequent activation of ERK1/2 and Akt. (supported by HL073890 and HL080402).
The involvement of PPARα in epoxide‐induced activation of adenosine receptors in angiogenesis
Authors: Rizvi Yasmeen Q , Choi Myung , Mehta Chander , Oyekan Adebayo
Published: June 2021

The role of peroxisome proliferator‐activated receptor alpha (PPARα) in angiogenesis is controversial. However, many studies showed that epoxyeisatrienoic acids (EETs), downstream mediators of adenosine A2B receptor (A2BR) and PPARα ligand are proangiogenic. Using the Zebrafish (Danio rerio) embryo, we here propose that EET‐induced PPARα activation is involved in A2BR‐mediated angiogenesis under hypoxic condition. Angiogenesis was evaluated at 24, 48, 72 hpf using alkaline phosphatase activity (APA). Under normoxic conditions, APA increased in a time‐dependent manner at 24, 48 (132 %) and 72 hpf (200 %). Clofibrate (10–200 μM) or fenofibrate (10–50 μM); PPARα ligands, NECA (0.1–100 μM), a non selective adenosine receptor agonist, or hydralazine (0.5–1mM), inhibitor of degradation of HIF‐1α did not affect angiogenesis at any of the time points. Compared to normoxia, hypoxia for 24 h reduced angiogenesis by 85 % (p<0.01), but not at 48 or 72 hpf. Using transgenic Tg(fli‐1:EGFP) embryos and epifluorescence microscopy, angiogenesis increased under normoxic conditions in response to clofibrate (50 μM), NECA (10 μM) or hydralazine (0.5 mM), but not to fenofibrate. Epifluorescence microscopy but not APA data demonstrate the possible involvement of PPARα in angiogenesis in the Zebrafish embryo model. However, APA data do not support hypoxia‐induced angiogenesis in Zebrafish embryo.
Mechanisms of AVE3085 Against Asymmetrical Dimethylarginine‐induced Endothelial Dysfunction
Authors: Xue Hong‐Mei , He Guo‐Wei , Huang Jun‐Hao , Yang Qin
Published: June 2021

Asymmetrical dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase (NOS) that may induce endothelial dysfunction. We studied the effect and mechanism of AVE3085, a newly developed transcription enhancer of endothelial NOS (eNOS), on ADMA‐induced coronary endothelial dysfunction. Porcine coronary small arteries were studied for endothelium‐dependent and ‐independent relaxations. Protein expressions of eNOS and nitrotyrosine as well as eNOS phosphorylation at serine 1177 (p‐eNOSSer1177) and threonine 495 (p‐eNOSThr495) were determined. ADMA treatment for 1‐hr significantly decreased endothelium‐dependent relaxation to bradykinin (61.3±4.7% vs. 92.2±2.6%, EC50: −7.04±0.16 vs. −7.80±0.14 LogM, p<0.05) that was preserved by the co‐treatment of AVE3085 (86.7±3.2%, −7.67±0.22 LogM). ADMA markedly reduced the protein expression of eNOS and p‐eNOSSer1177 whereas increased the expression of p‐eNOSThr495 and nitrotyrosine (p<0.05). The decreased expression of eNOS and p‐eNOSSer1177 was reversed by AVE3085. AVE3085 also lowered the expression of p‐eNOSThr495 and nitrotyrosine (p<0.05). AVE3085 prevents ADMA‐induced coronary endothelial dysfunction. Enhancements of eNOS expression and activation as well as reduction of oxidative stress underline the protection of AVE3085.

Supported by Hong Kong GRF grants CUHK4651/07M & 4789/09M and CUHK direct grant 2041457.
Genetic inactivation of endothelial nitric oxide synthase enhances vasodilator effect of cilostazol
Authors: Santhanam Anantha Vijay R , Hire Matthew G , Smith Leslie A , Katusic Zvonimir S
Published: June 2021

Cilostazol is an inhibitor of phosphodiesterase type III that possesses both antiplatelet and vasodilator effects. The present study aimed to determine the vasodilator effect of cilostazol in eNOS deficient (eNOS−/−) mice. Mouse aorta was contracted with sub‐maximal concentration of phenylephrine and concentration‐dependent relaxations to cilostazol (10−8–10−5 mol/L) were obtained in aortic rings, isolated from wild type and eNOS−/− mice. As previously reported, in aortas of wild type mice with intact endothelium, relaxations to cilostazol were significantly augmented as compared to aortas without endothelium. Surprisingly, genetic inactivation of eNOS significantly enhanced relaxations to cilostazol suggesting that chronic loss of nitric oxide may unmask compensatory vasodilator mechanisms significantly enhanced by the treatment with cilostazol. The augmentation of relaxations to cilostazol in aortas of eNOS−/− mice was endothelium independent and not inhibited by indomethacin, a cyclooxygenase inhibitor. Our findings support the view that patients suffering from vascular endothelial dysfunction induced by reduced availability of nitric oxide, may exhibit enhanced vasodilator reactivity to cilostazol.
Phosphorylation of endothelial nitric oxide synthase is increased in mouse model of GTP‐cyclohydrolase I deficiency
Authors: d'Uscio Livius V. , Katusic Zvonimir S.
Published: June 2021

Tetrahydrobiopterin (BH4) is an essential cofactor for endothelial nitric oxide synthase (eNOS). Suboptimal level of BH4 reduces formation of nitric oxide leading to uncoupling of eNOS. However, the exact molecular mechanisms underlying the effects of eNOS uncoupling on vascular endothelium in‐vivo are not completely understood. In the present study, we used the hph‐1 mouse, which displays GTP‐cyclohydrolase I deficiency. HPLC analysis revealed that BH4 levels were significantly reduced in the aortas of hph‐1 mice (3.9±0.5 pmol/mg; P<0.05 vs. wild‐type mice: 5.9±0.6 pmol/mg), while oxidative products of BH4, 7,8‐dihydrobiopterin (7,8‐BH2), were unchanged. Consequently, the BH4 to 7,8‐BH2 ratio was significantly reduced in the aorta of hph‐1 mice (P<0.05). Expression of eNOS protein was not different between wild‐type and hph‐1 mice. Unexpectedly, protein expressions of phosphorylated Ser1177‐eNOS and Ser473‐Akt1 were significantly increased in hph‐1 mice aortas (P<0.05). Consistent with uncoupling of eNOS, L‐NAME sensitive superoxide anion production and 3‐nitrotyrosine formation were significantly increased in aortas of hph‐1 mice (P<0.05). Protein expressions of CuZnSOD, MnSOD, and EC‐SOD were unaltered in the aortas of hph‐1 mice as compared with wild‐type mice. Our results suggest that uncoupling eNOS increases phosphorylation of Akt1 and eNOS. This effect is most likely mediated by increased formation of eNOS‐derived reactive oxygen species including superoxide anion and hydrogen peroxide.
Predictors of Measurement Error in Energy Intake During Pregnancy
Authors: Nowicki Eric M , Siega‐Riz Anna Maria , Herring Amy
Published: June 2021

Nutrition plays a critical role in maternal and fetal health; however, research on measurement error of energy intake during pregnancy is limited. Data were analyzed from 1,023 women with singleton pregnancies in the Pregnancy, Infection, and Nutrition Cohort Study. Second trimester diet was assessed using a food frequency questionnaire (FFQ). Estimated energy requirement (EER) was calculated using IOM prediction equations with adjustment for energy costs during the second trimester. Implausible values for daily energy intake (EI) were determined using confidence limits of agreement for EI/EER. Prevalence of low energy reporting (LER) and high energy reporting (HER) was 31.8% and 12.9%, respectively. In a multivariable analysis, pre‐gravid BMI was related to both LER and HER; LER was higher in both overweight (OR=1.96, 95% CI=1.26, 3.02) and obese women (OR=3.29, 95% CI=2.33, 4.65) than in normal weight counterparts. Other predictors of LER included marriage and higher levels of physical activity. HER was higher among subjects who were underweight, African American, less educated, and had higher depressive symptom scores. Nutrient density intakes for fat, calcium, riboflavin, and vitamin C differed between LER and non‐LER subjects. LER and HER are prevalent during pregnancy. Identifying their predictors may improve data collection and analysis methods to reduce systematic bias in reproductive studies.

Grant Funding Source: ASN
EGCG reduces Ang II‐induced NAD(P)H oxidase and NO synthase expression in HUVECs
Authors: Ahn Hee Yul , Kim Chan Hyung , Davidge Sandra T
Published: June 2021

Atherosclerosis is a chronic systemic disease of the vasculature with an inflammatory component. The impairment of vascular endothelial cell function which is an important determinant of atherosclerosis progression is in part due to increased release of reactive oxygen species (ROS). Angiotensin II (Ang II) causes endothelial dysfunction by increasing NAD(P)H oxidase‐mediated vascular superoxide production. Epigallocatechin‐3‐gallate (EGCG), the major bioactive polyphenol present in green tea and a potent antioxidant, has been reported to lower the risk of cardiovascular diseases such as atherosclerosis and hypertension. In this study, we investigated the effects of EGCG on Ang II‐induced expression of NAD(P)H oxidase and nitric oxide synthase (NOS), which may contribute to oxidative stress. We observed that Ang II stimulated mRNA and protein expression of NAD(P)H oxidase subunit and iNOS in human umbilical vein endothelial cells (HUVECs). EGCG inhibited Ang II‐induced expression of NAD(P)H oxidase subunit and iNOS in a concentration‐dependent manner. However, EGCG did not affect Ang II‐induced ROS production. Surprisingly, EGCG itself produced ROS in HUVECs. These results suggest that anti‐atherosclerotic activity of EGCG is likely not due to decreased ROS formation although there may be some benefit through inhibition of expression of NAD(P)H oxidase and NOS induced by Ang II.