bluuebunny/crossref_metadata_2025 huggingface dataset:
Structural properties of the extracellular domain of the synaptic adhesion protein α‐neurexin
Authors: Miller Meghan , Mileni Mauro , Harel Michal , Nakagawa Teru , Taylor Palmer
Published: June 2021

The neurexin (NRXN) family constitutes a class of membrane proteins ubiquitously expressed at pre‐synaptic glutamatergic and GABAergic terminals. NRXNs help maintain synaptic function and integrity through trans‐synaptic associations with neuroligins. Studies indicate that genomic aberrations play a role in mental disorders, including autism. NRXNs are expressed by 3 genes, each with 2 promoters corresponding to the α‐ or β‐NRXNs. α‐NRXNs differ from β‐NRXNs in their extracellular domain, which contains an additional 5 independently folding LNS domains and 3 EGF domains. Each LNS domain is structurally homologous to the extracellular domain of β‐NRXN. Neuroligins, α‐dystroglycan, neurexophilin, and the spider toxin α‐latrotoxin, are binding partners of 2 of the 6 LNS domains of α‐NRXN. Structural and functional roles of the remaining domains are unknown. By negative‐stain single particle EM we obtained a low resolution (~20 Å) structure of the extracellular domain of α‐ NRXN, providing a description of the conformation and insights into regions of flexibility. These insights enabled us to pursue higher resolution studies. From these data we present a model of the extracellular domain of α‐ NRXN that shows the structural arrangement of the domains, as well as the accessibility of known sites of protein interaction.

R37‐GM18360 and PO1‐ES10337
SIN‐1 cause downregulation of sirtuin expression in mouse astrocytes
Authors: Xie Qian Reuben , Shao Jiaxiang , Ying Weihai , Xia Weiliang
Published: June 2021

Astrocytes play essential roles in mediating inflammation in ischemic brain diseases. Sirtuins are important proteins regulating multiple biological processes including aging, inflammation and survival. The expression and function of sirtuins in astrocytes have not been reported. In this study, we use primary cultures of mouse astrocytes to study the expression of sirtuins, and how their levels respond in various conditions including oxidative stress induced by 3‐morpholinosydnonimine (SIN‐1), resveratrol, sirtinol, or pre‐treated with resveratrol or sirtinol followed by SIN‐1. Mouse astrocytes express all seven forms of sirtuins with sirt2, sirt3, sirt7 being the highest expressed. SIN‐1 (0.2–1 mM) profoundly down‐regulated the expression of all forms of sirtuins. Sirtinol (50–200 uM) significantly down‐regulated the expression of all forms of sirtuins except sirt3. Resveratrol (50–150 uM) did not significantly affect the expression of sirtuins. However, pretreatment of reseveratrol (100 uM) but not sirtinol attenuated the effect of SIN‐1 in downregulating the expression of sirtuins. Our findings suggest oxidative stress and the modulation of sirtuins expression might be closely linked. (Supported by “Rising Star” Grant from Science and Technology Commission of Shanghai and Young Investigator Grant from National Natural Science Foundation of China to W. X.)
Long‐term potentiation (LTP) induces co‐localization of calmodulin and neurogranin in the dendritic spines of mouse hippocampal CA1 pyramidal neurons
Authors: Huang Kuo‐Ping , L Freesia
Published: June 2021

Calmodulin (CaM) and neurogranin (Ng) are abundant neuronal proteins in the forebrain and their interactions have been hypothesized to enhance the synaptic plasticity. Deletion of Ng gene in mice causes deficits in learning the hippocampus‐ and amygdala‐dependent behavioral tasks, and the high‐frequency stimulation (HFS)‐induced LTP. Immunohistochemical staining of CaM and Ng in the hippocampal slices of wild type mice revealed that both proteins were co‐localized in the soma and dendrites of the principle neurons. However, unlike the relatively even distribution of Ng in the soma and dendrites of the CA1 pyramidal neurons, CaM exhibited an asymmetrical distribution in these neurons where the somatic concentration was at least ten times greater than that in the dendrites. Surprisingly, majority of the somatic CaM was localized in the nucleus. Thus, under the basal conditions the overwhelmingly high Ng/CaM concentration ratio in the distal dendrites renders very little free CaM for the activation of CaM‐dependent enzymes. In these slices, HFS (a single train of 1 s, 100 Hz) caused mobilization of CaM from soma to associate with Ng in dendrites that lead to the expression of LTP. After HFS, CaM exhibited a patchy distribution in the apical dendrites of the stimulated CA1 region. Both CaM and Ng was found to associate with the dendritic spines adjacent to the stimulating electrode. These findings suggest that the association of CaM and Ng at the stimulated dendritic spines may enhance the synaptic efficacy by increasing the Ca2+ transients as predicted by the “mass‐action mechanism” and they may also serve as synaptic tags for the stimulated dendritic branches. (Supported by Intramural Program of NICHD, NIH).
Effect of Giardiasis on Vitamin A Status in Mexican School Children
Authors: Lopez‐Teros Veronica , Valencia‐Juillerat Mauro E , Sotelo‐Cruz Norberto , Vazquez‐Ortiz Francisco , Rascon‐Duran Lucila , Tortoledo‐Ortiz Orlando , Pacheco‐Moreno Bertha Isabel , Mendoza‐Bermudez Diana , Astiazaran‐Garcia Humberto
Published: June 2021

Objective

Vitamin A deficiency (VAD) is a nutritional problem affecting the health of people in developing countries because VAD compromises innate and adaptive immunity, increasing a person's predisposition toward infectious diseases. In addition, a high prevalence of infectious diseases continues to be a problem in developing countries, including Giardia lamblia. Giardiasis may be related to VAD because of its ability to change the intestinal architecture, thereby compromising the absorption of vitamin A. The aim of this study was to evaluate the effect of giardiasis on vitamin A status in school children.

Methods

Thirty Giardia‐infected school children participated in this study. Vitamin A status was evaluated at baseline during active giardiasis and six months after anti‐parasitic treatment was administered. In addition, anthropometric and dietary data were collected.

Results

According to anthropometric indicators (age‐appropriate Z scores for weight, height, and body mass index) the children had a normal nutritional status. Although mean dietary vitamin A adequacy was >100%, only 73% (n=22) of subjects had an adequate daily vitamin A intake. There was a significant improvement on serum retinol levels once giardiasis was treated, leading to a 64.6% reduction of subclinical VAD (P<0.02).

Conclusion

Giardia lamblia has a negative effect on vitamin A status of Mexican school children, increasing their predisposition toward infectious diseases.
LACTATIONAL IRON DEFICIENCY; PERSISTANCE OF ANEMIA AND CELLULAR RESPONSES TO IRON REPLETION POST WEANING IN SPRAGUE DAWLEY RATS
Authors: Hegde Narasimha , Jensen Gordon L , Unger Erica L
Published: June 2021

Iron deficiency during infancy and its sustained effects on cognition and behavioral functions in later childhood have been well documented. Rodent models indicate that gestational/lactational iron deficiency reduces brain iron, an effect that persists into adulthood when iron supplementation begins at weaning. The molecular mechanisms related to persistent low brain iron in this model remain poorly understood and in part depends on iron delivery from peripheral iron stores to the brain. In this study, rats were fed iron sufficient or deficient diets from G15 until P21, and then maintained on an iron sufficient diet until P75. At P75, iron supplementation was unable to restore spleen and liver iron to control levels. This observation paralleled increases in hepcidin expression in liver and an upregulation of Nfkbia, Hif1á, and IL‐6 expression in spleen. These data indicate that altered transcription factor and inflammatory cytokine expression may play a role in the continued deficiency in iron homeostasis in iron supplemented rats. The molecular mechanisms for this disturbed iron regulation after supplementation is currently unknown, but these data begin to provide an explanation for these novel observations. This work is supported by the USDA National Institute for Food and Agriculture (2007‐35200‐18231).
Isatis indigotica induces hepatoma cell apoptosis via caspase‐independent pathway in vitro and in vivo
Authors: Chiang En‐Pei Isabel , Chung Ying‐Chun , Tang Feng‐Yao , Chung Chia‐Hua
Published: June 2021

We explored the chemotherapeutic potential of an Asian biennial herbaceous Cruciferae Isatis indigotica and investigated the mechanism by which it inhibits tumor growth in vitro and in vivo. Chloroform extract from dried leafs (CEDLI) had the strongest anti‐tumor activity among all fractions examined. CEDLI supplementation (0.26% for 2 mo) inhibited pre‐developed xenografted hepatoma growth by 43 % without inducing toxicity. Tumor immunohistochemical analysis showed a 33% decrease in proliferation and a 3.6‐fold increase in cell apoptosis in vivo. CEDLI induced sub‐G1 cell cycle arrest and apoptosis by activating p53, inducing Bax and reducing Bcl‐2, which in turn caused mitochondrial stress and led to the release of mitochondrial apoptosis inducing factor into the cytosol, followed by its translocation into the nuclear, resulting apoptosis in hepatoma cell‐lines. We provide novel in vivo evidence that Isatis indigotica has antitumor activity against liver tumorigenesis, and the chemotherapeutic activity of Isatis indigotica on hepatoma was associated with a distinguished caspase‐independent apoptotic pathway. Results from this study provide the molecular basis for Isatis indigotica as a potential anticancer treatment against human hepatoma.
Influence of quercetin supplementation on disease risk factors in community‐dwelling adults
Authors: Knab Amy M. , Shanely R. Andrew , Henson Dru A. , Jin Fuxia , Heinz Serena A. , Austin Melanie D. , Nieman David C.
Published: June 2021

In vitro and animal data indicate quercetin has antioxidative and anti‐inflammatory functions and lowers disease risk factors, but data in humans is limited. This study investigated the effect of quercetin supplementation (500 mg/d, or 1000 mg/d) on disease risk factors in a large group of adults (n=1,002, 60% female, 40% male) varying widely in age and BMI. Randomized groups (placebo, PL; Q500; or Q1000) were supplemented for 12 weeks. Plasma quercetin, inflammatory markers (C‐reactive protein, CRP, and five cytokines), diagnostic blood chemistries, blood pressure, and blood lipid profiles were measured. No differences in blood chemistries were found except for a small decrease in serum creatinine and increase in glomerular filtration rate in Q‐500 and Q‐1000 groups compared to PL. A small but significant decrease in mean arterial blood pressure (MAP) was measured for Q500 and Q1000 groups compared to PL. The change in serum total cholesterol was different between Q500 and PL groups, and there was a small but significant decrease in HDL in Q1000 compared to PL. Change in IL‐6 differed slightly between Q1000 and PL groups (interaction effect, P=0.021), but change in CRP and other inflammatory measures did not differ between groups. Two high doses of quercetin had a negligible influence on disease risk factors, with the most notable finding being a decrease in MAP. Supported by grants from Coca‐Cola and Quercetin Pharma
Effects of a diet rich in almonds on low‐density lipoprotein cholesterol (LDL‐C), LDL‐C particle size, abdominal adiposity and vascular health
Authors: Berryman Claire E. , Bordi Peter L. , West Sheila G. , Fleming Jennifer A. , Kris‐Etherton Penny M.
Published: June 2021

Controlled feeding and free‐living clinical studies have been conducted to evaluate the effects of tree nuts on lipids and lipoproteins, major risk factors for cardiovascular disease (CVD). Notably, benefits were observed consistently in all population groups studied. Of the tree nuts, almonds in particular, are a good source of monounsaturated fatty acids, arginine, α‐tocopherol, magnesium and manganese. Thus, the purpose of this research is to build the evidence base for approval of FDA health claims related to almonds and CVD. A randomized, 2‐period, crossover controlled feeding study was designed to compare the effects of two blood cholesterol lowering diets that meet the AHA recommendations. The nutrient profiles of the control diet (59.3% CHO, 15.4% PRO, 25.2% FAT, no almonds/day) and almond diet (51.8% CHO, 16.7% PRO, 31.5% FAT, 1.5 oz. of almonds/day) are matched for PRO (15–17%), saturated fat (~7%) and cholesterol (<200 mg/day). The study population consists of two cohorts of 25 men and women (BMI 20–35 m/kg2) with moderately elevated LDL‐C (50–95th percentile). We hypothesize that a cholesterol‐lowering diet designed with almonds confers greater cardioprotective effects than a cholesterol‐lowering diet without almonds on the basis of the LDL‐C lowering response, effect on LDL particle size, abdominal adiposity and vascular health. This study is supported in part by the Almond Board of California.
Influence of dietary protein on fecal quality and gene expression of apoptosis‐ and immune‐related genes in the colon of Miniature Poodles and German Shepherds
Authors: Nery Joana , Leray Véronique , Biourge Vincent , Martin Lucile , Dumon Henri , Nguyen Patrick
Published: June 2021

Digestive tolerance vary among dog breeds, which could be partially attributed to higher colonic fermentation. Highly digestible protein sources, such as wheat gluten, could lead to lower colonic protein fermentation. Our aim was to assess the effect of dietary protein on fecal quality, and mRNA expression of genes involved in colonocyte apoptosis and mucosa immune responses.

Five Miniature Poodles (MP) and 6 German Shepherds (GS) were fed with diets varying in protein source and level: low‐protein wheat gluten meal (LP) and high‐protein poultry meal (HP). Feces were scored daily (1=dry; 5=liquid) and fecal moisture was analyzed. Colonic biopsies followed each 14‐d trial period. The mRNA expression of proteins involved in apoptosis (Bcl‐2, Casp3), and in mucosa immune responses (IL7, TLR2, TLR4) was analyzed by RT‐real time PCR.

Fecal score and moisture were higher in GS than MP, and when feeding HP diet. The expression of IL7 was higher in GS than MP (P < 0.05), and when dogs were fed with HP diet (P = 0.03). The TLR2 mRNA expression was higher in the proximal colon of GS than MP (P = 0.08), and when feeding HP diet to GS (P = 0.03).

Up‐regulation of immune‐related proteins could indicate a higher response to commensal bacteria, leading to a lower quality of feces. Diets containing low levels of highly digestible proteins could be an adequate nutritional strategy to improve fecal quality. Research supported by Royal Canin.
Protein source modifies bone health in adult rats
Authors: Thorpe Matthew P , Norton Layne E , Moulton Christopher J , Johnson Amy J Wagoner , Evans Ellen M , Layman Donald K
Published: June 2021

Studies of protein and bone health report variable results. Differential effects of protein sources may explain incongruities. In particular, portions of sulfur amino acids may impose a bone‐demineralizing acid load while portions of leucine may enhance anabolic balance.

We randomized 52 rats, 52d old, to semi‐purified, isonitrogenous diets (16% energy as protein, 54% carb) with protein isolate from soy, egg white, wheat or whey. Animals were sacrificed at 11 wk, and femurs excised for 3 point bend tests of mechanical strength and dual x‐ray absorptiometry.

Greater force was required to break the femur midshaft with whey (119±10 N) compared to soy (110±9 N, p=0.02). Bone mineral mass was also greater given whey (480±25 mg) vs. egg (466±14 mg) and soy (456±27 mg); as well as in wheat (478±25 mg) vs. soy animals (p<0.05 for all comparisons).

Protein source modified bone health in this animal model independent of total protein and energy intake. These observations are not adequately explained by a negative effect of sulfur in protein sources, which is low in soy (39.8 mEq potential acid per 100g protein) compared to whey (46.2) and wheat (69.4).

Supported by National Dairy Council and Egg Nutrition Center.

Grant Funding Source: National Dairy Council
Secondary analysis of food group intake in prostate cancer patients undergoing radiotherapy and nutrient supplementation
Authors: Bartalone Rachel L , Brown Jean K , Browne Richard W , Wilding Greg , Tyrpak Donna , Horvath Peter J
Published: June 2021

Diminished nutritional intake and declining appetite are common in cancer patients during therapy. Prostate cancer patients (53) undergoing radiotherapy had 3‐day diet/activity records taken at baseline (T1), during therapy with supplementation (T3) and post‐therapy with supplementation (T4). Patients were given a Placebo, a multivitamin/mineral Dietary Reference Intake (DRI), or DRI with additional antioxidants (Orthomolecular, OM). As previously reported total sugar, carbohydrate and overall energy intake (EI) decreased significantly with Placebo, not DRI or OM. Food groups and discretionary calories (DC ‐ saturated fat, alcohol and added sugar) were examined to understand the decreased EI. Intakes of milk, vegetable, and grains did not change. DC decreased in the Placebo group during radiation. At T4 oil intake increased in the DRI group, and Placebo vs. DRI. There was a trend for an increase in meat intake at T3 in all groups and for fruit at T4 in OM. In conclusion, the decrease in EI in the Placebo group may be due to DC. Radiation therapy did not affect most food group intakes, regardless of treatment. Supported by a grant from The National Cancer Institute #R21CA102391‐2.

Grant Funding Source: The National Cancer Institute #R21CA102391‐2
The Relationship between Breastfeeding and Childhood Leukemia Risk in Oman
Authors: Wali Yasser , Waly Mostafa , Al‐Mukhaini Yahya , Ali Amanat
Published: June 2021

Recent case‐control studies on breastfeeding and childhood leukemia risk indicated that long duration (>6 months) is associated with decrease risk of the disease. Most of these studies have been conducted in the west and the matched controls were not selected from the same family siblings of the studied cases, this excludes the genetic causes of leukemia.

We decided to conduct a case‐control study to investigate the relationship between breastfeeding and childhood leukemia risk in Oman where breastfeeding is culturally favorable for a longer period of time (up to 24 months).

All of the recently diagnosed cases and registered cases at the national registry during (1999–2009) has been screened and a total number of 70 cases were recruited for this study. For each case, a matched control for age and sex was selected from the same family siblings and the ratio of cases to controls was 1:1.

Data of this study revealed that for breastfeeding duration of (6–12 months), 21% of cases were reported as compared to controls (12%). Meanwhile longer period of (12–24 months) was observed for 75% of cases as compared to controls (81%). For a period of (≥ 24 months) the percentage of cases was 4% as compared to controls (7%). All results were not statistically significant different (Chi‐square, χ2= 3.816), P>0.05.

Our study indicates that breastfeeding duration is negatively associated with childhood leukemia risk and other environmental and genetic factors might be synergizing for the disease occurrence in Oman.
Longitudinal associations of physical activity and cancer mortality ‐ the Third National Health and Nutrition Examination Survey
Authors: Parekh Niyati , Lin Yong , Vadiveloo Maya , Fitzgerald Nurgul , Lu‐Yao Grace
Published: June 2021

Background

Previous studies suggest that physical activity (PA) may afford protection against cancer risk.

Methods

This study evaluated longitudinal associations between PA and cancer mortality in the nationally representative Third National Health and Nutrition Examination Survey (NHANES III; 1988–1994), with an average follow‐up of 8.5y to mortality. Mortality was ascertained using National Death Index certificates. Metabolic equivalents (METS) were calculated from self‐reported PA collected in an in person‐interview. Cox‐proportional hazards models were used to compute hazard ratios (HR) for cancer mortality using SUDAAN in relation with level of PA.

Results

Any mention of PA vs. sedentary lifestyle was protective against cancer mortality (adjusted for age, race, sex, smoking, and BMI)(HR: 0.63, 95% CI: 0.47–0.84). High intensity PA was associated with further risk reduction (HR: 0.46, 95% CI: 0.27 – 0.78). Subjects in the highest vs. lowest tertile of METS had a 43% reduced risk of cancer mortality (HR:0.57, 95%CI: 0.41–0.80), p‐trend = 0.004. Leisure activity (gardening, dancing, yard work, and “other” active hobbies) was also associated with reduced risk of cancer mortality (HR:0.51, 95% CI:0.33–0.78).

Conclusion

In these preliminary analyses, all forms of PA confer protection against cancer mortality. PA should be considered as an integral part of cancer management.
Incorporation of conjugated linoleic acid into mouse tissues and the regulation of feed intake and body fat in the short term
Authors: Shelton Valerie J , Shelton Abigail G , Barnes Kimberly M
Published: June 2021

Conjugated linoleic acid (CLA) is known to decrease feed intake and body fat in mice. Our objectives were to determine the incorporation of CLA into tissues and the effect on serum hormones during the first 7 d of CLA consumption. Mice (n=150; 9‐wk‐old) were fed 0 (ad libitum or pair‐fed to CLA‐fed mice) or 2% CLA (ad libitum) for 1, 2, 3, 5, or 7 d. A body fat index was calculated; liver, adipose tissue, and brain were analyzed for fatty acid profiles; and serum insulin, leptin, and PYY were measured. CLA and pair‐fed mice ate less (P < 0.05) than control mice on d 1, 2, 3, and 7, but CLA‐fed mice were leaner than control and pair‐fed mice only on d 7 (P < 0.01). CLA was detected in adipose tissue starting on d 2 and in liver on d 1, but CLA was not detectable in the brain. Serum insulin tended (P = 0.11 and 0.13, respectively) to be increased in CLA‐fed mice on d 5 and 7, and serum leptin was non‐significantly reduced in CLA‐fed mice compared to control mice on d 7. The CLA‐induced reduction in feed intake was not sufficient to cause the loss of body fat as pair‐fed mice were not leaner than controls. The lack of CLA in the brain indicates that the effect on feed intake may be indirect.
Effect of a Short‐term Diet and Exercise Intervention in Children on Serum Lipomics and Markers of Metabolic Health
Authors: Izadpanah Ali , Barnard R. James , Burant Charles F. , Roberts Christian K.
Published: June 2021

The present study was designed to examine the effects of a short‐term diet/exercise program on markers of metabolic health and serum lipomics. 33 overweight children (15 boys, 18 girls, age 12.3±0.4 yr), were placed on an ad libitum, high‐fiber, low‐fat diet and daily exercise (2–2.5 hr) regimen in the Pritikin Longevity Center 2‐wk program. Fasting serum was taken pre‐ and post‐intervention for determination of glucose, lipids, metabolic risk markers, and lipomic analysis by gas chromatography. After 2 wks subjects lost weight (73.0±6.2 post vs. 75.7±6.5 kg pre) but remained overweight/obese (BMI: 27.2±1.7 vs. 28.2±1.8 kg/m2). Despite remaining overweight, no subjects had metabolic syndrome post‐ compared with 8 pre‐intervention. RHR and BP decreased significantly, p<0.005. Serum TG, Total‐C, LDL (p<0.0001), PAI‐1, resistin, leptin (p<0.001), HDL, IL‐8, amylin and insulin (p<0.05) decreased significantly. Serum IL‐10 (p=0.14) and IL‐1ra (p=0.29) increased and VEGF (p=0.07) decreased non‐significantly. Lipomic analysis revealed decreases in total lipids (79% of baseline), saturated fatty acids (14:0, 82%, 18:0, 87% and 20:0, 33%), and increases in 20:1 (125%), 22:6 (144%) and 18:1/18:0 (114%), all p<0.05. These results indicate changes in multiple indices of metabolic health and serum lipomics with short‐term, rigorous lifestyle modification, even in the face of remaining overweight/obese. This study was supported by a grant from the L‐B Research/Education Foundation and funding from UCLA.
Vegetarian diets are associated with a lower risk of metabolic syndrome. The Adventist Health Study 2
Authors: Rizzo Nico Samuel , Sabate Joan , Jaceldo Karen , Fraser Gary
Published: June 2021

Objective

To compare dietary patterns graded as vegetarian, semi vegetarian and non vegetarian in their relationship with metabolic risk factors and metabolic syndrome (MetS).

Design

Cross‐sectional analysis of 765 subjects (mean age 60 y) from the Adventist Health Study II calibration sub‐study. Type of diet was categorized based on food‐frequency questionnaire and classified as vegetarian (36%), semi vegetarian (17%) and non vegetarian (47%). ANCOVA was used to determine associations between type of diet and metabolic risk factors. Logistic regression analysis was used to calculate odds ratios (OR) for MetS risk. In all the analyzes adjustments were made for age, sex, ethnicity, socioeconomic status, alcohol intake, smoking, and physical activity.

Results

Vegetarians and semi vegetarians had significantly lower levels of fasting blood glucose, total cholesterol, waist circumference and BMI when compared to non vegetarians. Vegetarians had significantly lower levels of triglycerides, systolic and diastolic blood pressure. Vegetarians had the lowest risk for MetS, [OR 0.49, 95% CI 0.29–0.65, P<0.001] and semi vegetarians had a risk of [OR 0.59, 95% CI 0.38–0.94, P=0.025] when compared to non vegetarians.

Conclusions

Vegetarian and semi vegetarian dietary patterns are associated with more favorable levels of metabolic risk factors and a lower risk for MetS.
Pharmacological and toxicological evaluation of a synthetic analog of L‐carnitine on in vitro models
Authors: Gómez‐Solis Antonieta , De la Cruz‐Cordero Ricardo , Avalos‐Soriano Anaguiven , Duarte‐Vázquez Miguel Angel , Rodriguez‐Fragoso Lourdes , Reyes‐Esparza Jorge
Published: June 2021

The aim of this study was to evaluate the pharmacological and toxicological effects of a synthetic analog of L‐carnitine on in vitro models. For the study we used hepatic and a renal cell lines. For pharmacological evaluation were induced an in vitro model of insulin resistance. Cells were incubated in serum‐free DMEM containing either normal concentrations of glucose (5.5 mM D‐glucose) or high concentrations of glucose (30 mM D‐glucose) for 24 h. Cells were treated with L‐carnitine (2mM) and the analog (0.01, 0.1, 1, 10, and 100 uM, and 1and 2mM) and incubated for 24 h. Glucose and glycogen were determined in medium and cell extracts. For the toxicological evaluation were evaluated the viability and cell proliferation in hepatic and renal cells by using a MTT assay. The results showed that the analog reduced the glucose levels in conditioned media between 20–25% at concentrations of 100 uM, 1 and 2 mM, as compared with control group (p< 0.05). No changes were observed in intracellular glucose levels. However, an increase in the intracellular glycogen levels (100%) was observed. The analogue did no produce changes on cell viability or in cell proliferation in hepatic and renal cells. In conclusion, the synthetic analog is able to reduce the extracellular glucose and increase the intracellular glycogen levels. So, these results indicate that the analog might be used as l‐carnitine for treating the metabolic syndrome.
2‐ 15N Glutamine tracer overestimate rates of Arginine and Citrulline synthesis in healthy adult men
Authors: Tomlinson Christopher , Rafii Mahroukh , Ball Ronald O. , Pencharz Paul
Published: June 2021

Recent studies have documented transfer of labeled nitrogen from 2–15N glutamine (Gln) to citrulline (Cit) and arginine (Arg) in fasting human adults. Conversely, in neonates we have shown no synthesis of Arg from glutamate. Therefore, we performed a multitracer study with 2‐15N and 1‐13C Gln given enterally to 5 healthy men fed a standardized milkshake diet. There was no difference in plasma enrichments between the two Gln tracers. 1‐13C isomers of Cit and Arg were synthesized from 1‐13C Gln. Three isomers each of Cit and Arg were synthesized from the 2‐15N Gln tracer; 2‐15N, 5‐15N and 2,5 15N2. The enrichments of these isomers of Cit and Arg expressed as atom percent excess are shown in the table.

Enrichment of Arg and Cit isomers synthesized from 1‐13C and 2‐15N Gln

1–13C
2–15N
5–15N
2,5–15N

Cit (%)
1.41
2.61,2
4.02
0.8

Arg (%)
0.141
0.41
0.8
0.1

Significantly greater enrichment was found of both 5‐15N Arg and Cit compared to 2‐15N isomers2, indicating the amino N from Gln is mostly transferred to Arg and Cit by transamination. Similarly, the enrichment of the 1‐13C isomers was significantly less than the 2‐15N isomers 1 suggesting rapid formation of α‐ketoglutarate and recycling of the nitrogen label. Our results show Gln is acting primarily as a nitrogen donor for Arg synthesis and studies using 15N label overestimate Arg synthesis rates.

Grant Funding Source: CIHR
Effect of rapamycin administration in humans on the skeletal muscle protein anabolic response to essential amino acid ingestion
Authors: Dickinson Jared M , Drummond Micah J , Fry Christopher S , Glynn Erin L , Gundermann David M , Dhanani Shaheen , Volpi Elena , Rasmussen Blake B
Published: June 2021

Nutrient ingestion, in the form of essential amino acids (EAA), has been shown to concomitantly stimulate mTORC1 signaling and muscle protein synthesis (MPS) in humans. The goal of this study was to determine whether mTORC1 activation is required for the stimulation of MPS following the ingestion of EAA in humans. Assessment of mTORC1 signaling proteins were performed on muscle biopsies from the vastus lateralis of young (22±3 yr) individuals who were administered a single oral dose of a potent mTORC1 inhibitor (rapamycin) prior to ingesting 10 g of EAA. Preliminary analyses show little or no change in the phosphorylation status of mTORC1 and key downstream effectors, S6K1 and 4E‐BP1, from basal through 1 and 2 hours post EAA ingestion (P>0.05). In contrast, ingestion of 10 g of EAA in control subjects substantially increased phosphorylation of mTORC1‐associated proteins and MPS (P<0.05). These preliminary data suggest that administration of rapamycin attenuates the mTORC1 signaling response to EAA ingestion in human skeletal muscle and, therefore, the activation of mTORC1 may be necessary to fully stimulate MPS following ingestion of EAA.

Funding: NIH/NIAMS R01 AR049877 and NIA P30AG024832
Development and Evaluation of WillTry: An Instrument for Measuring Childrens Willingness to Try Fruits and Vegetables
Authors: Thomson Jessica Lee , McCabe‐Sellers Beverly J , Strickland Earline , Lovera Dalia , Nuss Henry J , Yadrick Kathleen , Duke Sara , Bogle Margaret L
Published: June 2021

This paper describes the development and evaluation of the WillTry instrument, a psychometric tool designed to measure childrens willingness to try fruits and vegetables. WillTry surveys were interviewer‐administered to 284 children in an elementary school and summer day camps located in rural Mississippi and Arkansas (United States) communities. Factor analysis was used to determine construct dimensionality. Additional evaluation included internal consistency, test‐retest reliability, and predictive validity. Factor analysis suggested a single dimension for the food items. The WillTry food scale had substantial reliability (intraclass correlation coefficients between 0.61 and 0.80) and sufficient internal consistency (Cronbachs alpha greater than or equal to 0.70). Results of the regression analysis for percent consumption of foods offered on WillTry response confirmed the predictive validity of the instrument. The results of these analyses provide psychometric evidence for the use of the WillTry instrument as a measure of willingness to try fruits and vegetables in rural, southern US children 5 to 14 years of age.
A pilot study evaluating the feasibility of a summer gardening program to prevent summer weight gain in overweight adolescents
Authors: Jacquart Sarah Rose , Schoeller Dale A , Adams Alexandra K , Larson Nathan , Dennis Samuel F , LaRowe Tara L , Carrel Aaron L
Published: June 2021

Background

Obesity in children is a growing problem in the US. Recent data shows that children experience greater weight gain during summer months than the school year, with overweight children experiencing the greatest weight gain.

Methods

The feasibility of an 8‐week summer gardening pilot program for overweight middle school children (11–14 years) to increase physical activity and fruit and vegetable (FV) consumption was evaluated. Six subjects were recruited to participate in 3 hours of gardening activities 5 days/week. Weight, energy expenditure (EE), and FV intake were measured.

Results

Program acceptance was evaluated using qualitative surveys and interviews. Mean weight change was −0.4 kg. Four subjects lost weight and 2 lost over 2.5 kg. Surprisingly, EE decreased. The study size was insufficient to detect significant changes; however, testing the acceptability of this type of program was essential. Qualitative data showed increased willingness to try new foods, liking at least one new food and a positive response to the program. Important logistic lessons learned were the importance of hiring a phlebotomist experienced in drawing blood from overweight children and the necessity of alternative activities to do on hot days. Future work is necessary to evaluate the effect of a larger gardening program on improving health outcomes. This research was supported by the Robert Woods Johnson Foundation.
The DIALBEST Type 2 diabetes peer counseling intervention: Process evaluation
Authors: Cruz Robert , Segura Sofia , Damio Grace , Perez‐Escamilla Rafael
Published: June 2021

These analyses include 60 participants that have completed the 12 month DIALBEST intervention that includes 17 home‐based peer counseling (PC) educational sessions in the topics of nutrition, diabetes and medication management. There were 930 PC contacts with patients with an average duration of 88 minutes per contact. The majority of sessions focused on nutrition (370) followed by diabetes complications (159). Participants evaluated the diabetes complications and food label sessions as very good/excellent (90%), the diabetes and medication management information as very good/excellent (88%), the portion size sessions as very good/excellent (85%), and the physical activity intervention as very good/excellent (83%). When asked about lifestyle changes, patients reported: eating out less (87%), eating less fast food (83%), planning their meals more (77%), being more physically active, routinely checking their feet (73%), eating lower fat foods and using food labels more (71%), as a result of their participation in DIALBEST. Funded by The Connecticut Center for Eliminating Health Disparities among Latinos (NIH‐NCMHD grant # P20MD001765).

Grant Funding Source: Nutrion (ASN)
Bid Regulates Murine Hepatocyte Proliferation by Controlling ER Calcium Homeostasis
Authors: Li Min , Ni Hong‐min , Baty Catherine J. , Ding Wen‐xing , Chen Xiaoyun , Yin Xiao‐ming
Published: June 2021

Bid, a BH3‐only Bcl‐2 family molecule, is generally known for its importance in activating the mitochondrial apoptosis pathway following death receptor engagement, particularly in hepatocytes. However, Bid also promotes hepatocyte proliferation during liver regeneration and carcinogenesis. This study aimed to determine the mechanism by which Bid regulated hepatocyte proliferation. Wild type and bid‐deficient primary hepatocytes were stimulated with serum and proliferation was measured by BrdU incorporation and immunoblot analysis of key cell cycle molecules. The hypothesis that Bid regulated endoplasmic reticulum (ER) calcium ([Ca2+]ER) homeostasis to affect proliferation was examined by measuring intracellular calcium level, by modulating the calcium level with pharmacological agents and by subcellular localization assay. We found that serum stimulated hepatocyte proliferation was dependent on calcium and depletion of calcium using thapsigargin or EGTA inhibited the proliferation. A portion of Bid was present in the ER and Bid negatively regulated [Ca2+]ER level, which was responsible for the delayed and reduced proliferation in bid‐deficient hepatocytes. Addition of ionomycin or reconstitution of Bid, particularly an ER‐targeted Bid, corrected the deficiency. In conclusion, we have revealed a novel regulatory mechanism of hepatocyte proliferation, exerted by Bid, a Bcl‐2 family protein otherwise known for its pro‐apoptosis activity under stress condition. This mechanism is mediate by the regulation on ER calcium homeostasis. We consider that this mechanism would be important for liver regeneration and carcinogenesis.
A protective function for interleukin 23 (IL‐23)‐IL‐17 axis in abdominal aortic aneurysm development
Authors: Xiong Wanfen , Knispel Rebecca , Baxter B. Timothy
Published: June 2021

Abdominal aortic aneurysm (AAA) is a multifactorial disorder in which T cell and macrophages infiltration is an important pathophysiological feature. Our previous studies have demonstrated that CD4+ cells and interferon gamma (IFN‐γ) play a critical role in the pathogenic mechanisms of AAA. The interferon 23 (IL‐23)‐IL17 axis have been linked to the pathogenesis of several chronic inflammatory diseases, including rheumatoid arthritis. However, whether IL23‐IL17 axis contributes to AAA formation is unclear. In the present study, we investigated the role of IL‐23 and IL‐17 in AAA formation. By studying a murine aneurysm model, we found that mRNA expressions of IL‐23 and IL‐17 were decreased in aneurysm tissue compared to normal aortic tissues. In addition, the response of IL‐23‐deficient mice to CaCl2‐aneurysm induction was tested. We found that relative to wild type control mice, the absence of IL‐23 resulted in larger aneurysms (P<0.05). Our data showed that IL‐23 deficiency increased MMP‐2 and MMP‐9 expression in AAA. The results demonstrate that IL‐23 may have a protective function in aneurysm development. This study demonstrates the pivotal role of lymphocytes in regulating matrix remodeling.
Monocyte Chemoattractant Protein‐1 and Interleukin‐8 Levels in Acute Inflammation Induced by Prolonged Exercise
Authors: Skenderi Katerina P , Papassotiriou Ioannis , Mitrakou Asimina , Mastorakos George , Anastasiou Costas A , Tsironi Maria
Published: June 2021

Monocyte chemoattractant protein‐1 (MCP‐1) and Interleukin‐8 (IL‐8) have important roles in the pathogenesis of many chronic inflammatory disorders. Our aim is to determine the association between MCP‐1 and IL‐8 levels and the degree of inflammation in athletes that participated in an ultradistance foot race. In 15 male athletes participating in the ultradistance foot race of the 246 Km (continuous, prolonged exercise for up to 36 hours), we measured circulating levels of MCP‐1 and IL‐8 before the race (phase I), at the end of the race (phase II) and 48h postrace (phase III). MCP‐1 levels at phase I (216.9±48.5 ng/L) (mean±SE), increased significantly at phase II (592.9±115.7 ng/L) and subsequently decreased at phase III (278.1±62.7 ng/L). At the same time period, IL‐8 followed a similar pattern (phase I: 9.4±4.5 ng/L, phase II: 28.5±8.8 and phase III: 8.9±4.3 ng/L). A significant positive correlation between MCP‐1 and IL‐8 was found at phase III (r=0.845, p<0.01), while this correlation was absent in the other two phases, indicating an independent response of each chemokine to inflammatory stimuli in each athlete. Prolonged exercise induces an inflammatory response that is expressed by an increase in circulating MCP‐1 and IL‐8 levels. Whether these changes have long‐term negative effects on the vasculature remains unknown.
Early detection of the acute phase response in sepsis by identification of small molecule metabolites in breath and biofluids
Authors: Butz Daniel E , Cook Mark E , Porter Warren P , Assadi‐Porter Fariba
Published: June 2021

The systemic inflammatory response associated with sepsis and septic shock causes significant morbidity and mortality in the critically ill, and early detection is critical to desirable outcomes. Septic shock is currently diagnosed using criteria such as body temperature and hypotension.

This study assessed the acute phase response to an infectious insult using breath 13CO2/12CO2 ratios (delta value) combined with metabolomic biomarker fingerprinting in serum.

Breath delta values drop 2–4 hours after endotoxin injection, whereas there is no breath response to dexamethasone or saline injected controls. In rat fecal peritonitis, the breath carbon delta value drops about 6 hours after injection while hypotension does not occur until 14 to 18 hours. Fingerprinting of small molecules in plasma by NMR spectroscopy indicates that concentrations of lactic acid, glucose, 2‐hydroxy‐butyrate, 3‐hydroxy‐butyrate, citrate, acetate, and ketogenic amino acids are increased. Similar metabolite profiles are exaggerated in plasma from ICU patients diagnosed with septic shock.

These data indicate that metabolic perturbations are evident at a very early stage of the acute phase response, and in the peritonitis model up to 12 hours prior to onset of septic shock. These technologies can provide an early warning for sepsis and may be used as a powerful diagnostic tool.

Supported by WARF and the Wisconsin Institute for Discovery.
Computational study on Trypanosoma brucei RNA editing terminal uridylyl transferase (TbRET2)
Authors: Demir Ozlem , Amaro Rommie E.
Published: June 2021

Trypanosoma brucei is the causative agent of African sleeping sickness, which is a major health problem in Africa. A large multiprotein complex, called the editosome, catalyzes a unique post‐transcriptional RNA editing in T. brucei and by this way, uridine nucleotides are inserted or deleted in mitochondrial pre‐mRNAs. RNA editing terminal uridylyl transferase 2 of T. brucei (TbRET2) is one of the key components of the editosome and is responsible for the uridine nucleotide addition. TbRET2 is shown to be vital for the bloodstream form of the parasite and no close human homologs are identified, which make TbRET2 a potential drug target. We will present insights about the structure and dynamics of TbRET2 based on our all‐atom molecular dynamics simulations of apo and UTP‐bound forms of the enzyme.
Evaluation of Angiogenic Activity in the Heart Using Isolated Primary Adult Cardiomyocyte Model
Authors: Konorev Eugene A , Leychenko Anna , Collier Eric , Matter Michelle , Kalyanaraman B
Published: June 2021

Cardiac pressure and volume overload is a potent stimulus for cardiac hypertrophy. The adaptive value of hypertrophic remodeling is largely determined by the ability of the heart to develop new vasculature, to accommodate the growth of myocardial tissue. This study focused on the production of proangiogenic vascular endothelial growth factor (VEGF) by cardiomyocytes. We cultured primary adult rat cardiomyocytes (ARCM) under hypoxic conditions, and collected culture media for VEGF measurements. Hypoxia caused a 3‐fold increase in VEGF release from ARCM. Primary adult mouse cardiomyocytes (AMCM) responded to hypoxia in a similar way. Hypoxia upregulated the expression of genes in AMCM that are controlled by hypoxia‐response elements. Expression of vegfa increased 5 to 8‐fold in hypoxic AMCM. Cobalt chloride, an agent that induces hypoxia‐like response, increased both VEGF production and vegfa expression in these cells. In separate experiments, mechanical stretch of ARCM caused a 3‐fold increase in VEGF release, as compared to non‐stretched cells. Thus, we conclude that cultured primary cardiomyocytes respond to hypoxia and mechanical stretch, factors that are present in overloaded hearts, with a significantly upregulated release of VEGF. The study suggests that the model may be valuable in designing agents that regulate the production of angiogenic factors by cardiomyocytes in the heart.
Inhibition Of Transmethylation (TM) Leads To Histone Modifications And Plays A Critical Role In The Epigenetic Regulation Of IL‐2 Gene Expression In Primary Human CD4+ T Lymphocytes
Authors: Patil Madhuvanti , Ghare Smita , Gobejishvili Leila , Joshi‐Barve Swati , McClain Craig J , Barve Shirish
Published: June 2021

IL‐2 is a critical lymphokine that exerts a wide spectrum of regulatory effects on the immune system. Recent work done in our laboratory has demonstrated that S‐adenosylmethionine (SAM) dependent transmethylation (TM) is essential for the function of CD4+ T lymphocytes including IL‐2 expression. SAM dependent TM impacts epigenetic changes involving histone modifications which play a key role in altering chromatin structure and transcriptional activity. The TM pathway can be significantly inhibited by the SAM metabolite S‐adenosylhomocysteine (SAH). Accordingly, the present work was carried out to examine the effect of decreased TM due to increased cellular SAH levels on IL‐2 gene expression in primary human CD4+ T cells.

Decreased SAM‐dependent methylation due to an increase in cellular SAH levels was achieved by treating the cells with an SAHH inhibitor, 3‐deazaadenosine (DZA). Examination of histone modifications occurring due to decreased SAM‐dependent methylation showed a decrease in the acetylated state of histone H3 at the IL‐2 proximal promoter region which is associated with the induction of gene transcription and expression. These data strongly support the notion that histone H3 acetylation is a critical epigenetic modification that regulates IL‐2 gene expression in primary human CD4+ T cells.
Evaluation of the Selective A2A Receptor Antagonist, SCH 412348, and the D2/D3 Receptor Agonist Pramipexole in Rat Models of Psychotic and Obsessive Behavior
Authors: Hodgson Robert , Bleickardt Carina , LaShomb Abigail
Published: June 2021

Dopamine D2/D3 receptor agonists, used to treat Parkinson's Disease (PD), have serious side effects, including hallucinations and obsessive behavior. Adenosine A2A receptor antagonists have been proposed as a PD treatment, but little has been done to evaluate potential psychiatric side‐effects. We tested the D2/D3 receptor agonist pramipexole, and the selective A2A receptor antagonist SCH 412348 in rodent prepulse inhibition (PPI). Pramipexole (0.3–3 mg/kg) induced a significant PPI deficit in rats and mice. PPI is used to preclinically model psychotic‐like behavior. Ropinirole (1–30 mg/kg) and SCH 412348 (0.3–3 mg/kg) had no effect. Currently there is no preclinical model of the obsessive behavior side effect associated with D2/D3 receptor agonists. To address this, we trained water‐deprived rats that licking a water spout would result in a mild footshock. We hypothesized that pramipexole‐treated animals would favor the rewarding stimulus over the punishing stimulus manifested as increased licking relative to vehicle‐treated animals. The resembles the clinical phenomenon in which individuals treated with a D2/D3 agonist appear to become hypersensitive to the rewarding effects of behaviors, but discount the negative consequences. Pramipexole (3 mg/kg) significantly increased licking; neither SCH 412348 nor ropinerole did. This represents a novel animal model of pramipexole‐induced obsessive behaviors. Taken together, these data indicate that treatment with an A2A receptor antagonist will not have the psychotic or obsessive side effects associated with pramipexole.