bluuebunny/crossref_metadata_2025 huggingface dataset:
Reliability and validity of a wearable heart rate and respiratory rate sensor system
Authors: Tharion William J. , Buller Mark J. , Karis Anthony J. , Mullen Stephen P.
Published: June 2021

Purpose: To determine if heart rate (HR) and respiration rate (RR) of the wearable Hidalgo Vital Sign Detection System (VSDS) were reliable and valid.

Methods: 8 men performed activities of varying intensity; low (e.g., lying prone & supine) medium (e.g., treadmill walking) and high (treadmill running). VSDS HR and RR were compared to reference standard (RS) HR (Schiller AT‐6 ECG) and RR (SensorMedics 2900 metabolic cart). Reliability was assessed by (1) mean between trials and (2) intraclass R. Validity was assessed by (1) mean between VSDS and RS, (2) Pearson r, and (3) Bland‐Altman (BA) plots. Acceptability of VSDS of HR measures was set a priori to be within 2 bpm (low activities) and 6 bpm (high activities) and RR to be within 1 breath per min (brpm) (low activities) and 3 brpm (high activities) of RS measures.

Results: HR showed no significant differences between trials. Intraclass R's ranged from 0.60 to 0.97; R for all activities: 0.88 (p<0.01). There was no significant difference between VSDS and RS HR. RR reliability showed no differences between trials except for pushups (mean Δ = −3.1 brpm; p<0.05). Intraclass R's ranged from 0.52 to 0.97; R for all activities: 0.90 (p<0.01). RR validity estimates showed differences between VSDS and RS (range: 1.9 brpm to − 10.4 brpm). Pearson r values ranged from 0.23 to 0.82.

Conclusion: VSDS HR data was both reliable and valid. RR was reliable but did not meet the validity criterion. This work was supported by U.S. Army Medical Research and Materiel Command.
A Na‐dependent fructose co‐transporter in lobster hepatopancreas
Authors: Ahearn Gregory A. , Sterling Kenneth M. , Cheeseman Christopher I.
Published: June 2021

3H‐Fructose (Fru) and 3H‐Glucose (Glc) transport were determined in brush‐border membrane vesicles (BBMV), baso‐lateral membrane vesicles (BLMV) and isolated cells (E, R, F, B) of lobster, Homarus americanus, hepatopancreas (HP). Glc transport was Na‐dependent in BBMV and not affected by a ten‐fold excess of Fru, indicating the presence of an apparent SGLT‐1‐like transporter. Glc uptake by BLMV was facilitative and was not inhibited by a ten‐fold excess of Fru. Fru uptake by BBMV was hyperbolic, Na‐dependent, and not inhibited by Glc. Fru uptake by BLMV was Na‐dependent and not inhibited by Glc. Hepatopancreatic E, F and B cells showed Na‐dependent uptake of Fru, while R cells did not. Na‐dependent Fru uptake by E cells was not inhibited by Glc or mannose (Man). Glucose uptake by E cells was facilitative. Western blot analysis of BBMV, BLMV and E, R, F and B cells with antibodies for mammalian GLUT‐2, GLUT‐4, GLUT‐5, SGLT‐1 and SGLT‐4 indicated the presence of the respective lobster HP orthologous proteins. A GLUT‐2‐like protein (∼50 kD) was present in all four cell types and only in BLMV. A GLUT‐4‐like protein (∼50 kD) was localized to BLMV. A 50 kD, GLUT‐5‐like protein, was present in F and B cells and in BLMV. An SGLT‐1‐like protein (∼75 kD) was detected in R, F and B cells; the signal was strongest in F cells. The SGLT‐1‐like protein was limited to BBMV. An SGLT‐4‐like protein was present in E, R, and F cells; the strongest signal was from E cells. Comparison of Fru and Glc uptake by BBMV, BLMV, and cells, indicated distinct Na‐dependent cotransporters for each sugar.

Supported by NSF grant IBN04‐21986.
Central neostigmine administration reverses alcohol‐ and hemorrhage‐induced hypotension
Authors: Mathis Keisa W. , Molina Patricia E.
Published: June 2021

The hemodynamic and neuroendocrine counteregulation to hemorrhage (HEM) is severely compromised during acute alcohol intoxication (AAI). We hypothesized that restoration of the neuroendocrine response would improve hemodynamic counter‐regulation in AAI + HEM. Previous studies showed that intracerebroventricular (ICV) choline (acetylcholine precursor) administration produced a transient activation of sympathetic nervous system outflow insufficient to improve MABP following AAI + HEM. We determined whether enhancing acetylcholine availability by ICV neostigmine (acetylcholinesterase inhibitor) would improve MABP following AAI and HEM. The dose response to ICV neostigmine (0.1–3 μg) was established in chronically‐catheterized, conscious male Sprague‐Dawley rats (225–250g) and 1 μg was selected based on its ability to produce an immediate (10 min) and sustained (2 h) increase (20%; P=0.07) in MABP. ICV neostigmine reversed the 12% (P=0.001) drop in MABP caused by alcohol (2.5 g/kg, 30% v/v) administration within 30 min, completely reversed the hypotension produced by 40% total blood loss, and improved 7‐day survival (100% vs. 25%) from HEM. These results demonstrate that ICV neostigmine reverses hemorrhage‐ and alcohol‐induced hypotension and produces a more sustained elevation in MABP. Therefore, we speculate that ICV neostigmine will improve compensatory responses to HEM during AAI. Supported by DOD PR‐054196 and NIAAA‐AA7577.
A novel prolactin receptor (PRLR2) in tilapia (Oreochromis mossambicus) is involved in the sea water acclimation response
Authors: Fiol Diego Fernando , Sanmarti Enio , Lim Andreana , Kültz Dietmar
Published: June 2021

A novel prolactin receptor (OmPRLR2) was identified and cloned in the euryhaline fish Oreochromis mossambicus using the suppression subtractive hybridization approach based on its response to hyperosmotic stress. OmPRLR2 protein has 55% identity with a known tilapia PRLR and 50% identity with human PRLR. Expression of OmPRLR2 was detected mainly in osmoregulatory organs (gills, intestine and kidney) but also in brain and muscle. Expression of the two tilapia PRLR differs in response to osmotic stress; OmPRLR1 expression is augmented during hypoosmotic stress but OmPRLR2 expression increased during hyperosmotic stress. Expression levels and tissue distribution of OmPRLR2 protein were quantified using specific antibodies, confirming its overexpression during hyperosmotic stress in gill cells. Stably transfected HEK‐293 cell lines overexpressing Tet‐inducible OmPRLR1 and OmPRLR2 were established. Receptors were functionally characterized using a casein promoter reporter system and through the analysis of expression of different prolactin (PRL) and growth hormone (GH) responsive genes. Specific responses of tilapia PRL177, tilapia PRL188, ovine PRL and tilapia GH were analyzed and show strong specificities for the ligands and resulting in activation of different signal transduction pathways. Additionally, cell lines expressing OmPRLR2 have increased tolerance to hyperosmotic media. Our results indicate that OmPRLR2 is a novel receptor contributing to osmosensory signal transduction in euryhaline fish.

This work was supported by a grant from the National Science Foundation (IOB‐0542755).
Postprandial effects of iron supplementation provided either with a meal or between meals on iron status and oxidative stress
Authors: Yang Z , Lönnerdal B , Brown KH , Allen LH , Dewey KG
Published: June 2021

Iron deficiency is highly prevalent globally, but recent evidence indicates risks associated Fe supplementation, some of which may be ameliorated by giving Fe with food. We conducted a pilot study to assess postprandial effects on plasma Fe, non‐transferrin bound Fe (NTBI) and isoprostane (a marker of oxidative stress) of an Fe supplement given with or without rice porridge, using a cross‐over design with a one‐week washout period. Within 4 h after ingestion of 45 mg Fe as FeSO4, plasma Fe concentrations were significantly greater when given without a meal than with a meal (p=0.02), although the treatment x time interaction was not significant (p=0.36). The increase in plasma Fe was greater for subjects with low vs. high Fe stores (p<0.01), regardless of treatment. Plasma NTBI concentrations were marginally greater for subjects with high Fe stores than for those with low Fe stores (p=0.06), but they did not increase after ingestion of Fe supplements, with or without a meal. Plasma isoprostane concentrations did not differ by treatment (with vs. without a meal, p= 0.73) or increase after ingestion of Fe supplements (p= 0.53), indicating no acute effect on lipid oxidation after an oral Fe dose. We conclude that giving iron with a meal prevents excess iron uptake by subjects with high iron stores. Further research is needed to examine long‐term effects of this approach. Supported by Jastro‐Shields Research Award and USDA, ARS.
Pathobiology of ovarian epithelial cancers (OEC) in relation to age‐specific incidence rates (ASIR)
Authors: Grimley Philip M , Henson Donald E , Schwartz Arnold M , Anderson William F
Published: June 2021

The pathogenesis of OEC is being critically explored by molecular profiling. We propose that population‐based analyses of OEC can link traditional histopathologic subclassification to underlying molecular events. NCI Surveillance, Epidemiology, and End Results registries provided data for >35,000 cases of OEC from >14 % of the US population for the years 1992‐2004. When ASIR (as cases per 100,000 woman‐years) are analyzed by log‐log plots (LLP), the exponential graphs express cumulative age‐dependent events. Thus, similar graphical patterns suggest a common pathogenesis, while different patterns imply etiologic heterogeneity. Results: For serous OEC, trajectories of LLP, including ages of ASIR inflection (flattening), reproducibly discriminated between subsets of low and high pathologic grade. This appeared congruent with a dichotomy in oncogenesis (Singer et al. Am J Pathol, 2002). In contrast, ASIR for all grades of mucinous OEC showed parallel trajectories of LLP with late age inflections. For OEC recorded as endometrioid, clear cell, undifferentiated, papillary‐, adenocystic‐ or adeno‐ carcinoma, graphical patterns were inconsistent with those of the serous or mucinous subsets. Summary: LLP analyses of ASIR for classical OEC revealed distinct graphical patterns for high and low grade subsets of serous carcinoma. This accords with reported molecular signatures. Supported by USUHS and GWCI.
Long–Term Effects of High‐and Low‐Glycemic Load Energy‐Restricted Diets on Metabolic Adaptation and the Composition of Weight Loss
Authors: Das Sai Krupa , Gilhooly Cheryl H. , Golden Julie K. , Pittas Anastassios G. , Fuss Paul J. , Dallal Gerard E. , Saltzman Edward , Roberts Susan B.
Published: June 2021

The effects of high glycemic load (HG) and low glycemic load (LG) diets on resting metabolic rate (RMR) and body composition changes in response to caloric restriction (CR) remains controversial. Objective To examine the effects of two CR diets differing primarily in glycemic load on RMR and the % of weight lost as fat and fat free mass (FFM). Methods One year randomized controlled trial of HG and LG diets at 30% CR in 29 healthy overweight adults (mean±SD, age 35±5y; BMI 27.5±1.5 kg/m2). All food was provided for 6 mo, and self‐administered for the next 6 mo. Outcome measures included RMR, fat and FFM. Results There was no significant effect of diet on change in RMR after accounting for the decreases in body fat and FFM, at any time point (p=0.51 at 3 mo, p=0.56 at 6 mo and p=0.85 at 12 mo). In the subgroup of individuals who lost >5% body weight the LG diet group had a significantly higher percentage of weight loss as fat and a significantly lower percentage of weight loss as FFM compared to the HG group (p<0.05). Conclusions This study found no evidence for differential RMR response to weight loss between the HG and LG diets. However, for a given weight loss, the LG diet resulted in greater fat loss and less FFM loss.

Support NIH grants NGA‐3U01‐AG20480, K23DK61506, #H150001; USDA 58‐1950‐4‐401
Effect of short term supplementation with Lactobacillus acidophilus LAFTI® L10 on resistance to influenza infection in young and old mice.
Authors: Han Sung Nim , Thomas Adam , Peters Emily , Elliott James , Meydani Simin Nikbin
Published: June 2021

Impairment of the immune function in the aged animals contributes to a decreased ability to clear pathogens when infection occurs. Probiotics have been shown to improve the immune response in animal models. We, therefore, determined the effect of 8 wks of dietary supplementation with Lactobacillus acidophilus (LBA) LAFTI® L10 on lung viral titer and selected peripheral immune responses in young and old C57BL mice infected with influenza virus. Young (2–4 mo) and old (20–22 mo) male C57BL mice were fed an AIN 93 diet supplemented with or without 1 x 109 per day of L10 for 8 wks. Animals were intra‐nasally infected with influenza virus and euthanized at 2, 5, 7 days post infection to determine viral titer and immune response. NK activity significantly increased while lymphocyte proliferation decreased following influenza infection. Young animals had significantly higher NK activity, and lymphocyte proliferation and lower lung viral titers compared to the old mice. LAFTI® L10 supplementation resulted in a significantly higher production of IFN‐gamma in the non‐infected old animals. There was no significant effect of LAFTI® L10 supplementation on viral titer, INF‐gamma, and NK activity in the infected mice. In conclusion, 8 wks of LAFTI® L10 supplementation increased INF‐gamma production in non‐infected old mice, but had no effect on influenza viral infection. Supported by DSM Nutritional Products, Inc. and USDA (Contract #58‐1950‐7‐707).
Golden Rice Can Provide Vitamin A Effectively as Assessed Using Isotope Reference Method
Authors: Tang Guangwen , Qin Jian , Dolnikowski Gregory , Russell Robert M , Grusak Michael A
Published: June 2021

Golden Rice (GR) is enriched with β‐carotene (β‐C) genetically. To determine its vitamin A value, intrinsically labeled GR was obtained by growing plants in a nutrient solution containing 23% D2O. The GR β‐C was enriched with deuterium with the highest abundance isotopomer peak at M+9. Healthy adult subjects (n = 5, 2M and 3F, 41–70y, BMI at 22 – 29) were given a reference dose of an oil capsule of 1 mg 13C10 ‐ retinyl acetate together with 200 g cooked white rice and 10 g fat as breakfast of day 1. At day 8, steamed GR rice (130 to 200 g of cooked weight) containing 1 to 1.5 mg β‐C together with 10 g fat was given as breakfast. Serum samples were taken at 0, 3, 6, 9, 13 hours of day1, day 2 at 0, 5 and 11 hours, day 3 at 0 and 11 hours, day 8 at 0, 5, 8, 11, and 13 hours, day 9 at 0, 5, and 11 hours, day 10 at 0 and 11 hours, and fasting samples at days 11, 15, 18, 25 and 33 (0 hour = 8 AM). The samples were analyzed using GC/ECNCI‐MS for the enrichments of retinol with M+5 and M+10. By using the response to the dose of 13C10 ‐ retinyl acetate as reference, our results showed that the conversion efficiency of GR β‐C is 4.1 ± 1.5 to 1 by weight. Therefore, GR is one of the most bioavailable plant foods to provide vitamin A. (Funded by USAID, NIH DK60021, USDA 58‐1950‐7‐707, 1950‐51000‐065‐04, and USDA 6250‐2150‐042.)
Pubic hair pubertal stage is more strongly associated with female adolescent bone mass than breast pubertal stage in an Asian‐White population in Hawaii
Authors: Novotny Rachel , Vijayadeva Vinutha , Grove John , Daida Yihe , LeMarchand Loic
Published: June 2021

Pubertal maturation is important to development of bone mass and development of maximal bone mass is primary prevention for osteoporosis. Tanner pubic hair (PH) and breast (B) pubertal staging is a reference indicator of maturation for females. We examined differences between B and PH stages as indicators of bone mass among adolescent girls in the Female Adolescent Maturation Study in Honolulu. The study includes 107 Asian and White girls, who were measured for bone mineral content (dual‐energy x‐ray absorptiometry, DXA), calcaneal ultrasound, tanner stages, anthropometry, diet and physical activity. PH stage was predictive of increased bone broadband ultrasound attenuation and speed of sound (SOS) by calcaneal ultrasound, and DXA bone mineral density, and marginally for bone mineral content in comparison to B stage. Children with a larger proportion of Asian ethnicity had more bone with all measures. Calcium intake was also significantly associated with increased SOS and DXA measures. Physical activity was significantly associated with calcaneal ultrasound; BMI was significant for (BUA); and protein for (SOS). In sum, PH stage was more strongly associated with Bone Mass than was B stage. These findings suggest that an androgenic hormonal profile is especially important to acquisition of bone mineral during adolescence among Asian and White females. Funding: DOD, USDA, NIH/RCMI
Anthocyanin Bioavailability from Acute Cranberry Juice Consumption and Evidence of Effects on Endothelial Function in Patients with Coronary Artery Disease
Authors: Milbury Paul E. , Vita Joseph A. , Finnerty Kathryn A. , Holbrook Monika , Palmisano Joseph , Hamburg Naomi M. , Blumberg Jeffrey B.
Published: June 2021

Observational studies suggest that intake of flavonoid‐rich foods reduces the risk of cardiovascular disease. To test the hypothesis that cranberry anthocyanins are bioavailable and affect endothelial function, a pilot study was undertaken in 15 patients (age: 62±8 y) with coronary artery disease. Blood and urine were collected between baseline (0 h) and 4 h after consumption of 480 mL cranberry juice (54%; 835 mg total polyphenols). Marked inter‐individual differences in plasma anthocyanin pharmacokinetics (determined by HPLC) were observed with maximum concentrations detected between 1–3 h. Endothelium‐dependent flow‐mediated dilation (FMD) was measured in the conduit brachial artery by ultrasound and in small fingertip arteries by pulse amplitude tonometry (PAT). Cranberry juice increased FMD (7.7±2.9 vs. 8.7±3.1%; p≤0.01) and log PAT ratio (0.10±0.12 vs. 0.23±0.16; p≤0.001) at 0 and 4 h after juice consumption, respectively. Hyperemic flow and baseline arterial diameter remained unchanged. Thus, cranberry anthocyanins are bioavailable and endothelial function improves following consumption of cranberry juice in patients with coronary artery disease. Research support: Ocean Spray, Inc. and USDA 58‐1950‐7‐707.
Engineering Tryptophan Residues into Glyoxysomal Malate Dehydrogenase
Authors: Johnson Adam , Bell Ellis
Published: June 2021

Glyoxysomal Malate Dehydrogenase, which normally lacks any tryptophans, has been engineered to contain a single tryptophan in a variety of different locations both in or near the active site, at sites distant from the active site and at sites near the dimer interface. These mutants have been used to probe conformational flexibility in regards to local conformation changes around the engineered tryptophan residues. This represents a significant tool in the ability to ascertain conformational integrity in mutants and conformational changes induced by allosteric ligands binding to distinct sites in the oligomer. Specifically studied in here are two mutations one distant and one near the subunit interface, L229W and I319W respectively. Mutants were expressed and purified using Ni‐NTA Affinity chromatography. These mutants have been characterized by initial rate kinetics, MALDI‐TOF and fluorescence spectroscopy, and used in both guanidine hydrochloride unfolding experiments and KI collisional quenching experiments in the presence or absence of various coligands to probe differential effects of ligands on conformational accessibility and stability. Studies are underway to construct a “dead” tryptophan subunit with a native subunit to monitor if the tryptophans report on conformational communication across the subunit interface.

Supported by NSF Grant MCB 0448905 to Ellis Bell
Proteomic Approaches to Defining Avian Natural Killer Cell Surface Proteins
Authors: Zhang Lei , Katselis George S. , Goto Ronald M. , Goebel Thomas W. , Yokoyama Wayne M. , Lee Terry D. , Miller Marcia M.
Published: June 2021

Receptor/ligand interactions that dominate the control of natural killer (NK) cell activity likely differ in detail among vertebrate species as the result of natural selection. We are interested in innate immune responses in the avian species that serve as alternate hosts for pathogens causing human disease. We would like to link the challenge of pathogens to the genetics of NK cell activity. We are applying a largely unbiased high throughput proteomic approach to identify components of the surface of chicken NK cells. Our goal is to develop methods for preparing membrane fractions in abundance so that rare proteins, e.g. NK cell receptors, can be detected by mass spectrometry. We are comparing different fractionation methods for the capacity to provide membrane proteins from embryonic spleen cells that exhibit natural cytotoxic activity. We found it necessary to enhance the yield of membrane proteins since simpler methods are inadequate. We are now comparing methods incorporating fractionation by relative hydrophobicity and by shotgun glycopeptides capture. NK cell surface proteins identified by molecular biology serve as controls in evaluating purification.

Supported in part by NSF MCB‐0524167
Improved High‐Performance Liquid Chromatography Method with Diode Array Detection for Simultaneous Analysis of Retinoic Acid Isomers, Retinol, Retinyl esters, Vitamin E, and Selected Carotenoids in Guinea Pig Tissues
Authors: Schäffer Michael W , Roy Somdutta Sinha , Mukherjee Shyamali , Das Salil K
Published: June 2021

We report the validation of a versatile reversed‐phase gradient HPLC method allowing simultaneous detection of retinoic acid (RA) isomers (13‐cis, 9‐cis, all‐trans‐RA), retinol, retinyl esters (retinyl palmitate (RP), stearate and acetate as internal standard), tocopherols (T) (α,γ,δ) and selected carotenoids (lutein, lycopene, β‐carotene) in Hartley guinea pig (GP) tissues, using a diode array detector. Under the tested conditions, the method yielded low intra‐ and inter‐assay variability, with good recovery and sufficient sensitivity. Limits of detection, tested linear range and coefficient of correlation (ng injected; μg/mL; r2‐values, respectively) were: 13‐cis RA (0.451; 0.023–22.562; 0.9981), 9‐cis RA (0.5044; 0.022–21.617; 0.9974), all‐trans‐RA (0.4930; 0.018–17.609; 0.9964), retinol (0.8615; 1.436–14.359; 0.9916), RP (0.3587; 0.028–28.181; 0.9981), α–T (6.6214; 0.368–367‐857; 0.9996), γ–T (5.9224; 0.279–279.669; 0.9982), δ–T (4.2148;), lutein (0.1159; 0.009–9.273; 0.9711), lycopene (0.6975; 0.074–7.363; 0.9992), and β‐carotene (1.1847; 0.01–10.202; 0.9758). This method was used to quantify the cited analytes in several GP tissues. It should be noted that lutein, a dietary component, was detected in GP lung. The significance of the presence of lutein in lung remains to be elucidated. (Supported by US Army Grant W81XWH‐06‐2‐0044).
Age Increases Susceptibility to Salmonella typhimurium Infection in C57BL/6 Mice: Role of the Immune System.
Authors: Ren Zhihong , Gay Raina , Thomas Adam , Pae Munkyong , Wu Dayong , Logsdon Lauren , Mecsas Joan , Meydani Simin N
Published: June 2021

Aging is associated with a decline in immune function, which predisposes the elderly to higher incidence of infections. Enterocolitis caused by Salmonella typhimurium (ST) infection is one of the most common foodborne diseases in US. We used streptomycin‐pretreated C57BL mice infected with ST to study the age‐specific differences in resistance and early immune response to ST. After young and old mice were inoculated orally with 3×108 or 8 ×105 cfu of ST, old mice had significantly greater ST colonization in ileum, colon, peyer's patches, spleen and liver. Ex vivo production of TNF‐α and IFN‐γ from old ST infected mice was significantly less than those from young infected mice. Spleens and MLN from young mice had significantly higher percentage of NK cells and neutrophils in response to infection than old mice. Old mice, but not young mice infected with ST, had lower total number of intracellular IFN‐γ and TNF‐α producing Mφ than non‐infected mice. Young mice, but not old mice infected with ST, had higher total number of intracellular IFN‐γ producing NK cells and neutrophils compared to non‐infected mice. These data show that old mice are more susceptible to ST infection than young mice, which might be due to impaired IFN‐γ and TNF‐α production from NK cells, neutrophils and Mφ in response to infection. Further studies are needed to determine the underlying mechanisms of impaired IFN‐γ and TNF‐α production to ST with age. Support by USDA #58‐1950‐7‐707
Inhibition of Aromatase Activity by Tannins and Flavonoids from Sorghum bicolor
Authors: Dowd Christopher , Hargrove James , Hartle Diane , Greenspan Phillip
Published: June 2021

Aromatase catalyzes the conversion of testosterone to estradiol and is a target for chemotherapy of breast and prostate cancers. We compared in vitro inhibition of human aromatase and porcine α‐amylase by bran extracts from two sorghum cultivars that have similar phenolic levels but differ in tannin content. Tannin‐rich sumac sorghum inhibited α‐amylase more strongly (IC50 = 1.4 μg/mL) than did black sorghum (IC50 = 11.4 μg/mL), which lacks tannins. Sumac inhibited aromatase activity slightly more than did black sorghum (IC50 = 10.8 μg/mL v. 12.2 μg/mL). Bovine serum albumin (BSA), which binds tannins, reduced inhibition by sumac but not black sorghum in both enzymatic reactions. Sumac tannins, separated from other flavonoids on Sephadex LH‐20, inhibited both enzymes but showed reduced inhibition in the presence of BSA. Non‐tannin flavonoids from either cultivar had higher IC50 values for both enzymes than the tannins, and BSA had little effect on their inhibition. We conclude that aromatase is inhibited both by non‐tannin flavonoids and by the tannins of sorghum. Only non‐tannin flavonoids are expected to be bioavailable to prostate or breast tissue. (Supported by USDA Grant # 5854304363)
Chronic green tea consumption and adipose tissue aromatase ‐ relationship with adipose tissue remodeling
Authors: Monteiro Rosário , Assunção Marco , Andrade José Paulo , Neves Delminda , Calhau Conceição , Azevedo Isabel
Published: June 2021

Green tea dinking is being supported due to its anti‐obesity properties. In the current work, we intended to study the putative direct effects of green tea (GT) prolonged intake in the adipose tissue (AT) from different anatomical locations. Male Wistar rats (n=5) were fed for 6 months with GT as the sole liquid source and compared to water‐drinking controls. Aromatase expression was determined in subcutaneous and visceral AT by western blotting and plasma estradiol and testosterone levels were determined by RIA. Adipocyte size was measured in AT sections, proliferation determined by KI67 immunostaining and AT apoptosis assessed by TUNEL. GT treatment decreased weight gain and increased aromatase in scAT and vAT. Plasma estradiol levels, which were undetectable in controls, raised to 10.7±1.8 pg/ml in GT‐treated rats. AT of GT‐treated rats had more proliferating cells in both locations. GT downsized adipocytes and increased apoptosis in vAT. These results suggest that GT consumption increases AT estradiol synthesis and results in AT remodeling, favouring hyperplastic rather than hypertrophic AT development. In conclusion, the present study presents new mechanisms for GT effects on body weight and highlights the potential benefit of GT drinking in obesity management. Supported by FCT (POCTI, Feder, Programa Comunitário de Apoio, SFRH/BD/12622/2003 and SFRH/BD/19497/2004).
Cold and dehydration induce changes in expression of glyceroporin HC3 in three tissues from gray tree frogs Hyla chrysoscelis
Authors: Yaganti Sushmita , Pandey Ram Naresh , Frisbie James , Goldstein David L.
Published: June 2021

Gray tree frogs, Hyla chrysoscelis, are potentially subject to dehydrational challenges from evaporative water loss and from freezing. In anticipation of the latter, this species accumulates glycerol during cold acclimation. We hypothesized that expression of the AQP HC3, a protein from the glyceroporin family that transports both water and glycerol, is up‐ regulated in response to cold and dehydration. We used immunohistochemistry to compare the expression levels of aquaporin HC3 in liver, kidney and skin of control (hydrated, 23º C), cold (hydrated, 4º C) and dehydrated (loss of 20% of standard body mass over one week at 23º C) gray tree frogs. During dehydration, expression of HC3 was markedly increased in liver, kidney and skin compared with control tissues. In contrast, HC3 expression was diminished in kidney and skin from cold animals, but was highly expressed in cold liver compared with warm and dehydrated conditions. The up‐regulation in HC3 protein expression in cold liver may reflect the role of this organ in synthesis and release of glycerol, whereas up‐regulation in kidney and skin during dehydration may reflect a regulated role of the aquaporin in water conservation. Research supported by NSF IOB‐0517301.
Fiber capillarization is augmented in the gastrocnemius muscle of mighty mini‐muscled mice
Authors: Wong Lisa Elaine , Garland Theodore , Hepple Russ
Published: June 2021

House mice selectively bred for high voluntary wheel running display a mini‐muscle phenotype; they display twice the mitochondrial enzyme activities of normal in a muscle that has half the mass. To determine the structural alterations facilitating aerobic metabolism in these mini‐muscles, cross‐sections of the medial gastrocnemius of normal (n = 6) and mini–muscled (n = 6) mice were stained with LeadATPase and myofibrillar ATPase, and analyzed for fiber size, capillarization and fiber type. Specifically, we tested the hypothesis that mini‐muscled mice would have greater fiber capillarization. On average, mini‐muscled mice had smaller mean fiber cross‐sectional area, due in large part to the presence of many fibers with small size in the superficial region. In addition, mini‐muscle mice had higher capillary‐to‐fiber ratio and higher capillary density. When fibers were binned according to size it was revealed that it was primarily the normal‐sized fibers of the mini‐muscles that had higher capillarity. These results support the hypothesis that the mini‐mice have higher indices of capillarity, which may be important to facilitate the superior mass‐specific aerobic function of these muscles. This research was supported by NSERC and NSF grant IOB‐0543429.
Chronic hypoxia and hyperoxia alter the post‐hatching growth trajectory of the American alligator
Authors: Owerkowicz Tomasz , Hicks James , Elsey Ruth
Published: June 2021

Recent paleoatmospheric models suggest that over the past 550 million years ambient oxygen level fluctuated widely. Such environmental changes likely influenced animal life and the course of evolution – a drop in oxygen level from 30% to 12% correlates with the mass‐extinction event at the Permo‐Triassic boundary. To further understand how chronic hyperoxia and hypoxia affect the growth and physiology of contemporary vertebrates, we incubated eggs and raised hatchlings of the American alligator under hypoxia (12%), normoxia (21%) or hyperoxia (30%). All animals were kept at 30°C and fed ad libitum every other day. At hatching, there was no significant difference in gross, musculoskeletal and visceral morphology of alligators from normoxia and hyperoxia. In contrast, hypoxic animals were significantly smaller, and showed 45% larger hearts but 30% smaller lungs (organ wet mass/yolk‐free hatchling mass). Two months post‐hatching, growth rate (g/day) was slowest in hypoxia and fastest in hyperoxia. Interestingly, alligators incubated under hyperoxia and transferred to normoxia at hatching continued to grow as fast as animals remaining in hyperoxia. This suggests that exposure to hyperoxia during embryonic development may have permanently affected the alligators’ respiratory and/or metabolic control centers, subsequently altering their post‐hatching growth trajectory. Funded by NSF IOB 04445680 to JWH.
Probing the open complex contact of the discriminator region on T5 N25 promoter
Authors: Wiwczar Jessica M , Hsu Lilian M
Published: June 2021

Bacteriophage T5 N25 promoter is rate‐limited at the promoter escape step during transcription by E. coli RNA polymerase (RNAP) and undergoes extensive abortive initiation as a result. The high abortive potential of the N25 promoter has been attributed to RNAP binding to consensus‐like −35 and −10 elements via σ4 and σ2 domain contacts, respectively, to form a stable open complex. Recently, Haugen et al. (2006 Cell125, 1069) on the rrnB P1 promoter, which contains an 8‐bp discriminator (DIS) region, that an additional contact exists between the −7 nontemplate base when it is a G residue with the σ1.2 domain; this interaction resulted in longer open complex half‐life. These authors postulated that an equivalent contact would occur with a G residue 2 nucleotides downstream of the −10 box in other DIS regions. We set out to test this hypothesis on the N25 promoter, which has a 6‐bp DIS and a G at the −5 position, using a 6‐thioguanosine (S6G) crosslinking probe to investigate its open complex footprint. We found that the S6G residue at −5 did not crosslink to σ, but rather to β or β′ subunit. Experiments are ongoing to clearly delineate which. Our results suggest that the −7G base of rrnB P1 and the −5G base of N25 – although both are situated two nucleotides downstream of the −10 box – are not conformationally equivalent in different open complexes. Supported by an NSF grant (RUI #0418316) to L. M. Hsu.
Three different IgD cell populations in channel catfish, Ictaulurus punctatus
Authors: Edholm Eva‐Stina , Stafford James , Sahoo Manoranjan , Bengten Eva , Miller Norman , Wilson Melanie
Published: June 2021

IgM and IgD are currently the only immunoglobulin (Ig) molecules identified in catfish. While catfish IgM resembles mammalian IgM, catfish IgD is quite different. It is a chimeric molecule consisting of a rearranged VDJ region spliced to the Cμ1 domain, followed by seven Cδ domains and either a transmembrane or secreted tail. To study catfish IgD function, monoclonal anti‐Igδ antibodies were generated. Flow cytometry and cell separation techniques demonstrate the presence of three IgD+ populations in peripheral blood leukocytes (PBL). Two are B lymphocyte‐like populations with one expressing messages for the membrane forms of Igμ and Igδ (IgM+/IgD), and the other only expressing the membrane form of Igδ (IgM−/IgD+). Moreover, PCR protocols amplify appropriate rearranged Ig messages from both populations indicating that they contain functional B cells. In contrast to the situation in mammals where IgM−/IgD+B cells are rare, IgM−/IgD+ cells can represent as much as 60% of the total PBL in some catfish. The third population consists of IgM−/IgD+granular‐like cells of unknown origin that do not express Igδ message, but appear to be armed with exogenous IgD via an IgD‐binding receptor. At the present, the function of the granulocytes is unknown, however they degranulate in response to cross‐linking of the surface bound IgD. Supported by NIH grant R01 AI19530; USDA grant 2005‐01812.
Purchased ready‐to‐eat foods are positively associated with children's animal source food intakes in rural Ghana
Authors: Colecraft Esi K , Marquis Grace S , Sakyi‐Dawson Owuraku , Lartey Anna , Ahunu Ben , Butler Lorna M , Jensen Helen H , Reddy Manju B , Lonergan Elisabeth , Canacoo Emmanuel
Published: June 2021

Purchased ready‐to‐eat (RTE) foods or street foods are a common part of children's diets in many countries. These foods may contribute to improving children's diet quality. We assessed the relationship between consumption of RTE foods and animal source food (ASF) intake by 2‐ to 5‐y‐old children in rural Ghana. Interviewers collected data on purchase of RTE foods in the past week (N=454). Children's total ASF intakes also were recorded. Among caregivers who had purchased RTE foods (N=370) in the past week, 36 % (N=133) of them had obtained the foods specifically for their 2‐ to 5‐y‐old child. There were no significant differences in sociodemographic characteristics of caregivers who purchased RTE foods for children and those who did not with respect to years of education completed, weekly income, household wealth rank, marital status, and occupation. Children who consumed RTE foods were significantly more likely to have eaten livestock meats, organ meats, chicken, and eggs in the past week compared to children who did not consume any RTE foods (P<0.05). Mean ASF diversity score was significantly higher for children who had consumed RTE foods in the last week compared to children whose diets did not include RTE foods (5.3 ± 2.2 vs. 4.7 ± 2.3; P<0.02). Purchased RTE foods may be an important source of ASF in young children's diets in rural Ghana. GL‐CRSP funded in part by USAID, Grant # PCE‐G‐00‐98‐00036‐00.
Chinese red yeast rice food spice inhibits androgen‐dependent and –independent prostate tumor xenograft growth by inhibiting cholesterol biosynthesis in androgen‐dependent and independent prostate cancer
Authors: Hong Mee Young , Moro Aune , Zhang Yanjun , Seeram Navindra P. , Heber David
Published: June 2021

Large cohort studies demonstrate that users of cholesterol‐lowering drugs have a reduced risk of prostate cancer. Xenograft studies demonstrate that statins can inhibit prostate cancer (Pca) xenograft growth by depleting lipid raft cholesterol. Chinese Red Yeast Rice (RYR) is a food spice containing a family of monacolins one of which is identical to lovastatin (LV). We have previously demonstrated inhibition of cell proliferation with RYR treatment in human prostate cancer cells in vitro. The present study examined the effects of RYR on growth of androgen‐dependent and androgen‐independent human prostate cancer xenografts in SCID mice. LNCaP and androgen‐independent LNCaP‐AR cells were inoculated subcutaneously in mice receiving LV or 5% RYR diets over 8 wks. RYR inhibited both androgen‐dependent and .independent prostate tumor volume by more than 60 % in SCID mice (P<0.01). RYR showed a more potent effect in reducing tumor size than LV. RYR diet produced lower serum cholesterol and prostate specific antigen levels compared to control and LV in LNCaP‐AR xenograft mice (P<0.05). RYR food spice and nutritional strategies for lowering cholesterol are planned to explore the anti‐cancer activity of RYR and statins in humans. Supported by UCLA CNRU CA 42710 and DOD W81XWH‐07‐1‐0158
Effect of fructose‐feeding on intestinal and renal expression of genes involved in fatty acid synthesis
Authors: Douard V. , Gurunathan A. , Muduli A. , Ferraris R.
Published: June 2021

Fructose is a potent stimulator of liver lipogenesis but little is known about the regulation of small intestinal (SI) genes involved in fatty acid synthesis, even though the SI contributes 10–40% of total body triglycerides. We pair‐fed two groups of 16 d old rats high fructose or glucose (control) solutions for 16 consecutive d (16F or 16G), and another two groups for 8 d followed by 8 d without sugars (8F or 8G) to evaluate the long‐term effects of fructose on the regulation of fatty acid metabolism. The expression of the fructose transporter GLUT5 was high in the liver, SI and kidney but only in the SI did fructose markedly upregulate GLUT5. In the liver but not in the SI of 16F fed rats, expression of lipogenic enzymes (FAS, HmGCoA, and Malic Enzyme 1) and the triglyceride transfer protein MTP were much greater than that in 16G, 8F and 8G. In the SI, withdrawal of sugar feeding in 16F and 16G markedly inhibited the expression of Malic Enzymes 1 and 3, MTP and HmGCoA in comparison with 8F and 8G. Expression of these genes in the kidney was not affected by sugar consumption regardless of feeding duration, indicating that lipogenic genes cannot be regulated in organs not involved in fatty acid synthesis. In contrast, genes involved in fatty acid synthesis are specifically stimulated by fructose in the liver but inhibited by both glucose and fructose in the small intestine. Support: NIH RDK075617A, NSF IOS‐0722365
Permeability increase induced by light‐dye treatment
Authors: Fu Bingmei M , Zeng Min , Liu Qin
Published: June 2021

To investigate the effect of light‐dye treatment on permeability of mammalian microvessels, we measured hydraulic conductivity (Lp) of the post‐capillary venules of rat mesentery in vivo under various treatments. We perfused a single microvessel with sodium fluorescein (0.5mg/ml in 1%BSA mammalian Ringer). After control measurement, we exposed the vessel to the laser (495 nm, 2.2 mW), in 2–5 min after the laser treatment, Lp increased from a mean control of 0.84 (± 0.17 SE, n = 8) to a mean of 1.26 (± 0.09 SE) × 10−7 cm/s‐cmH2O, a 1.9 fold increase. Longer time exposure to the laser (up to 30 min) did not increase Lp further. To decrease the increased Lp by the light‐dye treatment, we pretreated the microvessel with a 2 mM cAMP analog, 8‐bromo‐cAMP, for 20 min. Lp decreased from a mean control of 1.40 (± 0.12 SE, n = 5) to a mean of 0.93 (± 0.08 SE) × 10−7 cm/s‐cmH2O in 20 min. Unexpectedly, cAMP did not decrease the Lp increase induced by the laser treatment. Instead, after 2–5min exposure to the laser, Lp increased to a mean of 4.13 (± 0.17 SE) × 10−7 cm/s‐cmH2O, a 3.6 (± 0.13 SE) fold increase from the control. We did not see the Lp decrease by cAMP in a longer time (30min) when exposure to the laser. One hypothesis for this is that the laser might change cAMP to cGMP, which was known to increase permeability. This work is in part supported by NSF.
Renal cyst fluid proteomics in autosomal dominant polycystic kidney disease (ADPKD)
Authors: Lai Xianyin , Bacallao Robert L. , Blazer‐Yost Bonnie L. , Hong David , Mason Stephen B. , Witzmann Frank A.
Published: June 2021

ADPKD is characterized by localized autonomous cellular proliferation, fluid accumulation within the cysts, and intraparenchymal fibrosis of the kidney. Little is known about the cyst fluid's protein composition. We hypothesized that the complex collection of cystic fluid proteins (cystic proteome) plabys a major role in cyst formation/maintenance and contains yet unknown diagnostic and mechanistic features that are common to all forms of PKD. We analyzed five kidney cyst fluids from four patients with ADPKD. Tryptic peptides from plasma‐protein immunodepleted (ProteoPrep®) and undepleted cyst fluid samples were analyzed by LC‐MS/MS. The proteins were identified by SEQUEST and validated via Trans‐Proteomic Pipeline. 391 proteins were identified with >90% confidence; 251 of them in undepleted and 362 in immunodepleted samples. Immunodepletion removed >98% of the cyst fluid protein. A surprisingly large and functionally diverse number of proteins common to all 5 cysts was identified. These proteins may be of mechanistic interest and include Ig γ κ and fragments; complement components; vitronectin; orosomucoid; prostaglandin D2 synthase; and vitamin D‐binding protein. Supported by AFOSR Grant FA9550‐06‐1‐0083.
Optic Nerve Re‐Growth in Danio rerio
Authors: Mosier Amanda L , Saul Katherine E , Koke Joseph R , Garcia Dana M
Published: June 2021

Regeneration in the central nervous system of teleost fish is well documented. A study using epifluorescence microscopy was conducted to correlate regeneration stages of severed axons of the optic nerve in adult zebrafish (Danio rerio) with analysis of changes in gene expression in the retina. Optic nerve injury was performed on wild‐type zebrafish which were then sacrificed at one day intervals for 10 days. DiI neurotrace paste was injected into the eye 24 hours prior to sacrifice. Fish were whole mounted leaving the retina, optic nerve and brain in situ. Regenerating axons were visualized by epifluorescence microscopy. Preliminary results indicate that after 3 days axons have crossed the lesion site and by 7 days have reached the optic tectum of the brain. This work supported by NSF IOB‐0615762.
Gastric Bolus Feeding Rapidly Stimulates Hepatic Protein Synthesis in Neonatal Pigs
Authors: Gazzaneo Maria Carolina , Suryawan Agus , Wilson Fiona A. , Orellana Renán A. , Kimball Scot R. , Nguyen Hanh V. , Davis Teresa A.
Published: June 2021

Growth and protein deposition rates are more rapid during the neonatal period than at any other stage of postnatal life. Feeding stimulates protein synthesis in the liver, as it does in other tissues of the neonatal pig. The purpose of this study was to examine the feeding‐induced time course of the changes in protein synthesis, the proportion of ribosomal protein (rp) mRNAs in polysomes, and translation factor phosphorylation in liver. Neonatal pigs (n=36; 5‐7 day old) received a gastric bolus meal and were sacrificed at fasting and after 30, 60, 90, 120, or 240 min post‐feeding. Liver protein synthesis reached a maximum 30 min post‐feeding, was sustained through 90 min, and returned to baseline by 240 min. The proportion of rp (S4 and S8) mRNAs in polysomes increased within 30 min of feeding, remained high at 120 min, and returned to baseline value by 240 min. Phosphorylation of eukaryotic initiation factor (eIF) 4E binding protein (4EBP1) and rp S6 kinase 1 (S6K1) increased by 30 min, remained elevated to 120 min, and returned to baseline by 240 min. These results suggest that feeding induced a rapid movement of hepatic rp mRNAs into polysomes and increased neonatal liver protein synthesis by enhancing translation initiation. Furthemore, these increases were sustained for at least 90 min after a meal, but returned to baseline by 240 min. (Supported by NIH grants AR‐44474 and DK‐15658 and USDA/ARS 6250510000‐33).
Acute Effects of Enteral Leucine Supplementation of a Low Protein Diet on Muscle Protein Synthesis in Neonatal Pigs
Authors: Murgas Torrazza Roberto , Suryawan Agus , Orellana Renan , Gazzaneo Carolina , Nguyen Hanh , Almonaci Rose , Davis Teresa
Published: June 2021

Protein synthesis and eukaryotic initiation factor (eIF) activation are increased in skeletal muscle of neonatal pigs parenterally infused with insulin and amino acids (AA), particularly leucine. We hypothesized that enteral Leu supplementation of a low protein diets in neonatal pigs will acutely increase muscle protein synthesis to a rate similar to that achieved by feeding a high protein (HP) diet. Fasted 5‐d‐old piglets (n=12) were gavage fed at 0 and 60 min either: 1) low protein diet (LP), 2) low protein diet supplemented with Leu (LP+L) to equal HP diet, or 3) HP diet. Diets were isocaloric and lactose was equal. Fractional protein synthesis rates and translational initiation control mechanisms were examined 90 min after feeding. Muscle protein synthesis increased in the LP+L group compared to the LP group alone and did not differ from the HP group. LP+L and HP increased the phosphorylation of S6K1, eIF4G, and 4E binding protein‐1 (4E‐BP1), decreased inactive 4E‐BP1▸eIF4E complex abundance, and increased active eIF4E▸eIF4G complex formation in muscle. There were no difference between groups in PKB phosphorylation. Our results suggest that low protein diets supplemented with leucine stimulates muscle protein synthesis to a rate similar to that achieved by high protein diets and this stimulation involves activation of mTOR downstream effectors. (Supported by Ajinomoto Amino Acid Research Program and USDA 58‐6250‐6‐001).