bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma fatty acids, human milk fortification and lipid oxidation in premature infants
Authors: Friel James , Kim Jongsoo , Hossain Zakir , McKay Dylan , Diehl‐Jones William
Published: June 2021

Human milk fortifiers (HMF) are routinely added to mother's milk (HM) for the premature infant. We reported data on 65 premature infants (birth weight < 1500 g) who were either fed HM with varying levels of HMF or formula (F) while they were in hospital receiving oral feeds. A significant increase (P<0.05) in urine_F2‐isoprostanes was found as HMF supplementation increased (Pediatr Res. 2011 Feb; 69(2): 160–4). Isoprostanes are prostaglandin‐like compounds formed in vivo from the free radical‐catalyzed peroxidation of essential fatty acids primarily arachidonic acid (AA C20:4n6). Linoleic acid (LA 18:2n6) is a polyunsaturated fatty acid used in the biosynthesis of AA. LA in HM ranges from 7 to 24 % of total fatty acids. LA in HMF is 140 mg per packet. We hypothesized that premature infants who had higher (n=7, providing total samples 20) F2 isoprostanes in urine would have more LA in their plasma from BM + HMF than those who were lower (n=10, providing total samples 31). Plasma samples from infants at bi‐weekly visits were analyzed for total fatty acids by standard assay using GS‐FID and heptadecanoic acid as the internal marker. Results (%w/w) were: High; LA 21.3 + 1.6 vs low 23.8 + 0.8, P < 0.05) and AA: High; 6.5 + 0.3 vs Low 6.8 + 0.4 NS). The hypothesis is not supported and suggests that other reasons exist for elevated isoprostanes in premature infants fed HMF than plasma lipid levels. Supported by CIHR and MICH
Dietary‐resistant starch improves glycemic control in STZ‐induced diabetic rat
Authors: Huang Hui Yu , Chao Huan‐Chun , Kao Yu‐Yu , Tsao Shu‐Ping , Sie Chang‐Hong , Chen Wen‐Chi
Published: June 2021

Dietary resistant starch (RS) is dietary carbohydrates that resist digestion in the small intestine and reach the large intestine. In the current study, we hypothesize that supplement dietary resistant starch with meal can improve glycemic control in diabetic rat. The RS was added into diary high fat diet, and used STZ to induced hyperglycemia with the Sprague Dawley (SD) rats for 10 weeks. The High fat diet and the RS diet contained 7 % ( weight/weight) resistant starch. SD rats (n=40) were divided into four groups, and served as control, high fat‐fructose group (HFD), HFD+RS group. The body weight, food intake were monitored weekly. The blood glucose, OGTT, insulin level, and HbA1C were measured during or after treatment period. The result showed that RS reduction of postprandial and fasting blood glucose level, HbA1C and AUC. Also RS improved insulin sensitivity in the HFD+RS group as indicated by HOMA‐IR. In clusion, RS is beneficial to overcome from the high fat diet/STZ induced diabetic complicated.
Post‐feeding serum metabolic responses to extensively hydrolyzed whey (WPH) or native whey concentrate (WPC) in rats
Authors: Roberts Michael D. , Cruthirds Clayton , Lockwood Christopher , Pappan Kirk , Company Joseph M. , Brown Jacob E. , Booth Frank W.
Published: June 2021

The purpose of this study was to compare how feeding extensively hydrolyzed whey protein (WPH) versus its native whey protein concentrate (WPC) in 8‐h fasted rats transiently affects serum metabolic profiles. Male rats (~300 g) were fed an isonitrogenous amounts of WPH or WPC and sacrificed 15 min‐, 30 min‐and 60 min post‐gavage for serum extraction. Rats fasted for 8–9 h were used as controls (CTL). Serum biochemicals were detected using GC/MS and LC/MS/MS. 333 serum metabolites were detected amongst all groups. Both WPH and WPC increased several amino acids (1.2–2.8‐fold), branched chain amino acids (BCAAs, 1.2–1.7‐fold), and serum peptides (1.1–2.7‐fold) over the 60 min time course compared to CTL rats (p < 0.05). However, WPH feeding rapidly (at 15‐min post) increased select amino acids (including BCAAs) and peptides compared to WPC and CTL feedings (p < 0.05). WPH and WPC led to increases in numerous free fatty acids (FFAs) over the post‐feeding time course compared to CTL rats (up to 6‐fold increases, p < 0.05). However, WPH feeding uniquely resulted in a 30‐min post‐feeding elevation in numerous FFAs compared to WPC feeding (p < 0.05), which may be due the WPH‐induced increase in circulating epinephrine compared to CTL and WPC (p < 0.05). These data provide a unique postprandial time‐course perspective as to how WPH versus WPC feedings affect circulating biochemicals.

Supported by Bionutritional Research Group
Cholanemia induces skeletal muscle wasting despite stimulation of translation initiation, decreased autophagy, activation of Yes Associated Protein (YAP) and proteosomal signal activation in mice
Authors: Desai Moreshwar S. , Koo Sue Jie , Eblimit Zeena , Suryawan Agus , Coss‐Bu Jorge A , Nguyen Trung C , Davis Teresa , Orellana Renan A
Published: June 2021

Skeletal muscle wasting is a common co‐morbidity in cholestatic liver diseases. We hypothesize that cholestasis and bile acid (BA) excess (cholanemia) directly induces muscle atrophy by decreasing translation and stimulating protein degradation in muscle. Biliary fibrosis and cholestasis was induced in 6–8 week old male C57BL/6J mice by feeding a DDC supplemented diet for 3 weeks and compared with isocaloric chow fed mice (n=3/group). DEXA scan, exercise tolerance, and fasting circulating BA, glucose and insulin were analyzed, and quadricep muscle signaling was determined. Compared to controls, DDC had 1/3 less food intake, lower body weight (17±0.6 vs. 25±0.5g), lean (11.4±0.8 vs. 16.1±0.4g) and fat mass (2.3±0.1 vs. 3±0.2g) and demonstrated early fatigue on the treadmill. DDC had liver injury, cholestasis, cholanemia (BA: 1000±250 vs. 7±1 μmol/L), and lower glucose (60±5 vs. 150±15mg/dl) and insulin levels (0.15±0.01 vs. 0.53±0.01 ng/L) than controls. DDC had lower muscle mass (0.05±0.006 vs. 0.14±0.01g), higher phosphorylation of AKT, 4E‐BP1 and eIF4G, lower AMPK, eIF2α, YAP phosphorylation, and lower LC3‐II and MuRF1 abundance than controls. These findings suggest that cholanemia stimulates translation initiation, and represses autophagy and protein degradation signaling despite physical evidence of muscle atrophy. (Supported by Pediatric Critical Care Internal Funding)
Dyadic relationships between young athletes’ and parents’ competition goals and eating styles
Authors: Frith Amy , Kaye Miranda , Vosloo Justine
Published: June 2021

While parenting influences child eating behavior, our aim was to examine dyadic relationships between parent and young athletes’ competition goals and eating styles. In male and female competitive youth swimmers, 8 to 17 y (Mage = 12.30; SD = 2.70), and their parent (n=37 dyads), we assessed achievement goals with the 2×2 Achievement Goals Questionnaire for Sport: mastery approach (MAp) and avoidance (MAv), and performance approach (PAp) and avoidance (PAv) goals. Parent and swimmer eating styles were measured using Dutch Eating Behavior Questionnaire (DEBQ and DEBQ‐C) and Emotional Eating Scale (EES and EESC). We used Actor Partner Independence Models (APIM) in Mplus to examine dyadic relationships. For female swimmers, parental MAp, MAv and PAp goals were associated with swimmers’ DEBQ‐C scores (i.e., better eating style) (β = −0.54, p < 0.01; β = − 0.49, p < 0.01; β = −0.45, p < 0.05, respectively). For male swimmers, parental and own MAp goals were associated with swimmers’ DEBQ‐C scores (i.e., worse eating style) (β = 0.60, p < 0.01; β = 0.50, p < 0.05, respectively). Female (PAp) and male (MAv) swimmers’ goals were related to parental eating style (β = 0.61, p < 0.01; β = −0.59, p < 0.05). Patterns of association were similar for EES and EES‐C. Results suggest that parental competition goals more strongly influenced child eating styles than the child's own goals, yet young athletes also influenced parental eating styles.
PLASMA VITAMIN D LEVEL AMONG WOMEN WITH AND WITHOUT POLYCYSTIC OVARIAN SYNDROME
Authors: Abdi Jemilla , Redinger Derek , Broughton K Shane
Published: June 2021

Polycystic ovarian syndrome (PCOS) is an endocrine problem in women of reproductive age. While Vitamin D is involved in calcium and phosphorus homeostasis, evidence indicates an association between vitamin D deficiency and metabolic syndromes in PCOS.

This study evaluated plasma vitamin D levels in polycystic and non‐polycystic women following supplementation with long chain omega‐3 PUFA and Vitamin D3. Twelve polycystic and 8 non polycystic women consumed 2.3g n‐3 PUFA for three months and 1000IU of Vitamin D3 and n‐3 PUFA for an additional two months. Fasting blood samples were collected monthly with plasma Vitamin D levels assessed by HPLC.

Plasma Vitamin D3 was not affected by supplementation in either group nor was there a difference in plasma Vitamin D3 between PCOS and non‐PCOS at baseline. Plasma Vitamin D2 was 2‐fold higher in PCOS than non‐PCOS at baseline. Plasma vitamin D2 was reduced with exclusive n‐3 PUFA ingestion and further reduced with Vitamin D3 supplementation. However, plasma vitamin D2 levels remained over 2‐fold higher in PCOS than non‐PCOS throughout (P≤ 0.05). Vitamin D3 supplementation may not be critical for women with PCOS. However, as Vitamin D2 was significantly higher in polycystic versus non‐ polycystic women, and was responsive to n‐3 PUFA ingestion, D2 supplementation may value in both groups.
Inhibitory effect of novel red ginseng treated with high temperature and high pressure on oxidative damage
Authors: Yu Seok‐Yeong , Yoon Bo‐Ra , Hong Hee‐Do , Lee Young‐Chul , Kim Young‐Chan , Cho Chang‐Won , Kim Kyung‐Tack , Lee Ok‐Hwan
Published: June 2021

Ginseng is one of the most commonly used traditional herbal medicines. This study was to investigate the inhibitory effect of novel red ginseng treated with high temperature and high pressure in oxidative stress‐induced mouse by high intensity exercise. Experimental groups were no exercise group(CON), high‐intensity exercise group(E), common red ginseng extract supplementation with high‐intensity exercise group(CGE), novel ginseng red extract supplementation with high‐intensity exercise group(NGE). High‐intensity exercise was performed by treadmill running for two weeks. After the experimental period, changes of glutamic pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase(GOT) in serum was measured, and MDA levels, glycogen contents, glucose‐6‐phosphate dehydrogenase(G6PDH), lactate dehydrogenase(LDH) activities and gene expressions of Cu/Zn SOD and myogenic factor4(MRF4) on tissue were analyzed. Our results show that the levels of GOT, GPT and MDA in NGE group were significantly lower than those of other groups. Moreover, the levels of muscular glycogen, G6PDH activity, LDH activity of NGE group were markedly higher than those of other groups. In the case of gene expression on muscle tissue, CGE or NGE groups led to up regulation of SOD and MRF4 gene expression. These results indicate that NGE can inhibit high‐intensity induced oxidative stress and enhance exercise capacity.
Deletion of TNFα receptor ‐1 and 2 exacerbates trans‐10, cis‐ 12 CLA induced hepatic steatosis in the mouse
Authors: Hussein Mahmoud , Giesy Sarah , Bauman Dale , Boisclair Yves
Published: June 2021

Chronic intake of trans‐10, cis‐12 conjugated linoleic acid (t10, c12 CLA) causes detrimental effects such as lipid accumulation in liver and increased expression of the pro‐inflammatory cytokine tumor necrosis factor alpha (TNFα) in adipose tissue depots. TNFα is involved in dysregulation of hepatic lipid metabolism and insulin signaling. The role of TNFα in mediating the stimulatory effects of t10, c12 CLA on lipid accumulation in liver is unclear. To examine this issue, wild type C57BL/6J female mice (WT) and their counterparts lacking both TNFα receptor‐1 and 2 (TNFRKO) were fed diets containing 0.5% of either oleic acid or t10, c12 CLA for 2 weeks. As expected, t10, c12 CLA reduced adipose tissue mass and caused hyperinsulinemia and hepatic steatosis in WT mice. Hepatic steatosis in WT mice was associated with increased expression of many enzymes involved in lipogenesis. Surprisingly, the effects of t10, c12 CLA in TNFRKO liver were maintained for lipogenic gene expression and exacerbated for steatosis. We conclude that the effects of t10, c12 CLA on hepatic lipid metabolism do not require TNFα signaling.
Consumption of cranberry powder shifts urinary protein profile in healthy human subjects
Authors: Krueger Christian Gerald , Meudt Jennifer J , Howell Amy B , Khoo Christina , Shanmuganayagam Dhanansayan
Published: June 2021

We hypothesize that consumption of cranberry products containing A‐type proanthocyanidins (PAC) modulates expression of endogenous proteins by cells of the urinary tract. Urine samples at baseline and post 7 day consumption of cranberry powder (36mg PAC equivalent/day) were collected from ten human subjects. We performed liquid chromatography mass spectrometry (LC‐MS/MS) analysis on an LTQ Orbitrap to monitor changes in urinary proteome in response to cranberry powder consumption. This new method has advantages over the classical proteomic approach in that it allows for the detection and determination of relative ratios of a large number of proteins. Supplementation of cranberry powder resulted in statistically significant lowering of eight proteins/peptides in the urine. The identification of these proteins in context of cranberry powder supplementation serves as a novel discovery using a unique LD‐MS/MS methodological approach to assess shifts in urinary proteome. Many of these proteins seem to play regulatory role in immune processes and tumorigenesis. Additionally, the methodology utilized in this discovery phase only provides a semi‐quantitative assessment of the shifts in the urinary proteome that occurred. Support for this research was provided by Ocean Spray Cranberries, Inc. and Complete Phytochemical Solutions, LLC.
Cirsium setidens Nakai extract inhibits adipocyte differentiation and High‐Fat Diet‐induced obesity in Mice
Authors: Lee YoungJun , Cho Ju Hyun , Kim Bong Kyun , Choi Hyeon‐Son , Lee Boo Yong , Lee Ok‐Hwan
Published: June 2021

Cirsium setidens Nakai, a wild perennial herb, grows mainly in Kangwon province, Korea, and it has been reported to contain bioactive ingredients with various medicinal activities including treat edema, bleeding and hemoptysis. However, the potentiality of anti‐obesity effects of C. setidens Nakai has not been fully investigated. Thus, we investigated the anti‐obesity effect of C. setidens Nakai extract (CNE) on lipid accumulation in 3T3‐L1 cells during the differentiation and obesity in high‐fat diet (HFD)‐ induced obese C57BL/6 mice. Furthermore, evaluate the concentrations of serum lipids, including total cholesterol, triglycerides, HDL cholesterol, and LDL cholesterol, in mice fed a high‐fat diet. Our results show that CNE inhibited adipocyte differentiation of 3T3‐L1 preadipocytes. Exposure to 100 ¥ìg/mL, CNE significantly reduced lipid accumulation (~30%) in 3T3‐L1 cells compared to control cells by downregulating gene expression of adipogenesis‐related mRNA such as peroxisome proliferator‐activated receptor‐¥ã (PPAR‐¥ã). In addition, CNE supplementation in mice fed a HFD revealed that significantly decreased HFD‐induced increases in body and organs weights as compared to the HFD group. These results suggest that CNE can be useful in the treatment of obesity as non‐toxic natural resources.
Lutein and zeaxanthin for the prevention and treatment of age‐related macular degeneration (AMD): a systematic review and meta‐analysis
Authors: Vishwanathan Rohini , Berger Samantha , Johnson Elizabeth J
Published: June 2021

Lutein (L) and zeaxanthin (Z) are associated with AMD. The objective was to evaluate the evidence on dietary and supplemental L and Z in the prevention and treatment of AMD through a systematic review and meta‐analysis. Medline® was searched from inception to the 2nd week in February 2012 and abstracts were screened based on predefined eligibility criteria. Sixteen studies were included, 2 randomized controlled trials (RCTs), 7 prospective, 2 retrospective and 6 case‐control studies. One RCT showed 10 mg/d of L for 1 year significantly improved visual acuity (VA) in AREDS category 3 AMD patients. No improvement in VA was observed in the second RCT with up to 20 mg/d L for 6 months in AREDS category 2, 3 or 4 AMD patients. Two out of 7 prospective studies found increased intake of L and Z to be associated with reduced risk of AMD. The two retrospective studies did not find significant associations. Three out of 4 case‐control studies that evaluated serum L & Z did not find significant differences between AMD cases and controls. In one study, serum Z, but not L, was inversely associated with exudative AMD. Two case‐control studies that assessed dietary L & Z intakes found low intakes to be associated with higher odds of AMD. Meta‐analysis is currently being performed and will be crucial in drawing substantial conclusions. Long term L supplementation seems to be an effective treatment option for AMD. Support: Bausch & Lomb.
The Relationship of Lutein and DHA in Cognitive Function
Authors: Mohn Emily , Vishwanathan Rohini , Lichtenstein Alice H. , Matthan Nirupa , Hausman Dorothy B. , Woodard John L. , Davey Adam , Johnson Mary Ann , Poon Leonard W. , Green Robert C. , Gearing Marla , Johnson Elizabeth J.
Published: June 2021

Alzheimer's disease (AD) likely stems from many mechanisms, two of which are oxidative stress and inflammation. Lutein, an antioxidant, and DHA, an anti‐inflammatory fatty acid, are preferentially stored in neural tissue and are positively associated with cognition. While DHA decreases neuroinflammation, lutein prevents oxidation of DHA. DHA may also increase lutein in the brain by modulating levels of HDL, a main transporter of lutein. Therefore, the aim of this study was to determine if a synergistic relationship exists between brain levels of lutein and DHA and cognition. Sixteen cognitive tests were completed pre‐mortem by subjects from the Georgia Centenarian Study. Brain tissues from 48 decedents were analyzed for carotenoids and fatty acids using HPLC and GC, respectively. Regression analysis revealed a significant, non‐synergistic lutein/DHA interaction for 5 cognitive tests, 4 of which are from the Consortium to Establish a Registry for AD test battery. Eight significant interactions were found between the temporal and frontal cortices (P<.05). The results indicate that the association of lutein or DHA with cognition is dependent on the other; however, their combined effect is less than expected if added together. Mechanistic studies on lutein and DHA are needed to further characterize the relationship between these two nutrients. Support: NIH 1PO1AG1755, DSM, Abbott Nutrition, USDA (5819500014).
Inhibition of in vitro pancreatic cancer cell migration by circulating anthocyanins and their metabolites from plasma of human subjects
Authors: Kuntz Sabine , Kunz Clemens , Rudloff Silvia
Published: June 2021

Pancreatic tumor cell migration has important roles in cancer progression and is associated with poor prognosis. We investigated whether circulating plasma anthocyanins and their metabolites (PAM) deriving from fruit juice intake affect cell migration through modulation of the expression of matrix metalloproteinases MMP‐2 and MMP‐9. Ten healthy subjects consumed an anthocyanin‐rich fruit juice (840.9 mg/L). Before (PAM0min) and after (PAM60min) ingestion, blood samples were taken and PAM were prepared by centrifugation and subsequent solid phase extraction. PAM were subjected to the pancreatic cancer cell lines PANC‐1 and AsPC‐1 for 24 and 48 h. Finally, cell migration was determined in a Boyden chamber, the mRNA expression of MMP‐2, MMP‐9 as well as NF‐kB by real‐time PCR. Application of PAM60min lead to a reduced migration of PANC‐1 cells after 48h, whereas AsPC‐1 migration was not affected. Concomitantly, we observed reduced MMP‐2 and MMP‐9 as well as NF‐kB mRNA expression levels in PANC‐1. In contrary, AsPC‐1 showed lower level of NF‐kB which were not influenced by PAM60min. Also, MMP‐2 and ‐9 were not altered. Thus, it can be concluded that in dependency of the cancer phenotype anthocyanins and/or their metabolites may inhibit pancreatic cancer cell migration by reducing NF‐kB dependent MMP‐2 and ‐9 expressions in physiological concentrations.
Cardiometabolic risk factors are influenced by Stearoyl‐CoA Desaturase‐1 (SCD1) polymorphisms and n‐3 polyunsaturated fatty acids supplementation
Authors: Rudkowska Iwona , Julien Pierre , Couture Patrick , Lemieux Simone , Vohl Marie‐Claude
Published: June 2021

Polymorphisms (SNPs) in the stearoyl‐CoA desaturase‐1 (SCD1) gene may play a role in the development of the metabolic syndrome.

Objective

To determine if SCD1 gene SNPs are associated with cardiometabolic risk factors prior and after n‐3 polyunsaturated fatty acids (PUFA) supplementation.

Methods

210 subjects completed a 2‐wk run‐in period followed by 6‐wk supplementation with 5g of fish oil (1.9g EPA and 1.1g DHA). Risk factors were measured pre‐ and post‐ supplementation. FA composition of plasma phospholipids was analyzed by gas chromatography and the desaturase indices were calculated SCD16 (16:1n‐7/16:0) and SCD18 (18:1n‐9/18:0). Genotyping of 8 SNPs of the SCD1 gene was performed.

Results

N‐3 PUFA supplementation decreased plasma triglycerides (TG), SCD16 and SCD18 ratios with an increased plasma glucose levels. In repeated measures analysis of variance, where the effects of interest were SNP, supplementation and interaction between the SNP*supplementation on cardiometabolic risk factors, SNPs in SCD1 modified: TG (rs508384), interleukin‐6 (rs3071), C‐reactive protein (rs3829160) and SCD18 ratio (rs2234970). Interaction effects were observed for plasma glucose (rs508384) and insulin levels (rs508384 and rs3829160).

Conclusion

Cardiometabolic risk factors are modulated by genetic variations in the SCD1 gene alone or in combination with n‐3 PUFA supplementation.

Funding: CIHR.
Selection of reference gene for qPCR analysis of hypothalamic appetite regulation
Authors: Colley Danielle L , Castonguay Thomas W
Published: June 2021

Identification of an internal control gene for normalization is required for qRT‐PCR experiments. Vandersompele et al (2002) have demonstrated the need for housekeeper genes that are unaffected by experimental treatments. Several investigators have selected β‐actin as the preferred housekeeper in examining the effects of sugars on hypothalamic appetite regulatory genes. We set out to determine if access to different sugar solutions could alter hypothalamic gene expression in rats. Presented are three sets of observations related to the selection of a housekeeper. First β‐actin varied across treatments, making it a poor choice. Second, a poorly selected housekeeper can result in reporting of false conclusions. When fold change was calculated using β‐actin, differences in AGRP were observed among treatment groups. However, when analyzed with RPLP as housekeeper no differences were observed. Third, sometimes using a poor housekeeper does not lead to false conclusions. Differences in some genes were significant independent of the housekeeper. Here we demonstrate that housekeeper selection can impact conclusions. Supported in part by a grant from the Maryland Agricultural Experiment Station.
Association of dietary and supplemental folate intake and polymorphisms of the MTHFR, MTHFD1, and TYMS genes and colorectal cancer in a population‐based study
Authors: Ashmore Joseph H. , Berg Arthur , Gallagher Carla , Miller Paige , Muscat Joshua , Zhu Junjia , Hartman Terryl , Lazarus Philip , Lesko Samuel
Published: June 2021

We evaluated the role of dietary and supplemental folate and 40 polymorphisms from three genes involved in folate‐mediated one‐carbon metabolism (FOCM) on colorectal cancer (CRC) risk in a population‐based case‐control study in Pennsylvania. Cases and controls reported dietary intakes and supplement use for the year before diagnosis or interview, respectively, via a modified food frequency questionnaire. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using unconditional logistic regression to assess the effect of folate from the diet and supplements and from FOCM genes (MTHFR, MTHFD1, and TYMS) on CRC risk among 686 incident cases and 740 controls. After multivariate adjustment, CRC risk was not significantly associated with dietary or supplemental folate intake. However, compared to no use, folate supplement use was inversely associated with colon cancer risk (OR = 0.77; 95% CI = 0.59–1.00). In preliminary analysis, only the rs17824591 MTHFD1 polymorphism was significantly associated with CRC risk (OR per allele = 1.21, 95% CI = 1.01–1.45). Further investigation of gene‐environment interactions is needed to elucidate the impact of diet and FOCM‐related genes on CRC risk. Partial support by the Pennsylvania Department of Health – Grant #4100038714.

Grant Funding Source: Pennsylvania Department of Health
Sexual dichotomy in calcium buffering capacity may be dependent on the severity of endoplasmic reticulum stress in the skeletal muscle of the vitamin D3 deficient transgenic G93A mouse model of amyotrophic lateral sclerosis
Authors: Shahsavar Shayan , Taheri‐Shalmani Siavash , Solomon Jesse Adam , Gianforcaro Alexandro , Hamadeh Mazen Jamal
Published: June 2021

Background

We previously demonstrated that dietary vitamin D3 (D3) restriction hastened the decline in functional capacity following disease onset in the G93A mouse model of amyotrophic lateral sclerosis (ALS). ALS is a progressive neuromuscular disease characterized by the degeneration and death of motor neurons.

Objective

In this pilot study, we analyzed the quadriceps of G93A mice following dietary D3 restriction at 2.5% the adequate intake for vitamin D receptor and markers of intracellular calcium trafficking (calbindin d28k, calretinin), endoplasmic reticulum (ER) stress (SERCA2, CHOP) and apoptosis (bax/bcl2 ratio, caspase 12 cleaved/pro ratio).

Methods

Beginning at age 25 d, 42 G93A mice were provided food ad libitum with either adequate (AI; 1 IU D3/g feed; 12 M, 11 F) or deficient (DEF; 0.025 IU D3/g feed; 10 M, 9 F) D3. At age 113 d, the quadriceps were analyzed for protein content. Differences were considered significant at P ≤ 0.10.

Results

In females, DEF had 30% lower calbindin d28k (P = 0.075), 69% higher cleaved/pro caspase 12 ratio (P = 0.100), and 103% higher bax/bcl2 ratio (P = 0.057) vs. AI. DEF males had 67% higher CHOP (P = 0.048) vs. AI.

Conclusion

Dietary D3 at 2.5% the AI reduces calcium buffering capacity and increases prolonged ER stress‐induced apoptosis in female, but not in male, G93A mice. We hypothesize a reciprocal interaction between ER stress and calcium buffering. (Supported by NSERC and Faculty of Health, York University)

Grant Funding Source: NSERC and Faculty of Health, York University
The structure of associated diseases in children with alopecia areata
Authors: Dzhetpisbaeva Zulfiya , Abduldayeva Aigul
Published: June 2021

The aim of our study was to investigate the interdependence of the severity degree of alopecia areata and comorbidity in children under the age of 14 years. We observed 17 children aged 1 to 15 years suffered from alopecia areata. Paraclinical examination included general clinical, clinical and biochemical laboratory tests, ultrasound study of the internal organs. History of disease revealed in 2 children (11.8%) food allergy, 3 (17.6%) children had the bottle‐feeding since a birth day. Various factors played a provocative role in the occurrence of relapses and severity of process. Thus, more than 50% of these were chronic foci of infection (early caries, chronic tonsillitis, dysbiosis), somewhat less deferred protracted psycho‐emotional disorders (stuttering, fear, insomnia, cries at night), mainly between the ages of 1,5 to 5 years and in children of 7–9 years of old. One quarter of the patients had colds and their treatment. They were predisposing factors for development of severe alopecia areata and its recurrence. In assessing the severity of clinical symptoms by counting the percentage of hair loss, we obtained the following data: in 4 (23.5%) patients diagnosed the total form, and in 12 (70.6%) ‐ the various forms of alopecia with a number of local lesions from 1 to 5. Only 1 (5.9%) had a diffuse form of alopecia.
Evaluation of Macrophage Migration Inhibitory Factor Level in Sera of Patients with Alopecia Areata
Authors: Younan Doreen Assaad , Ebeid Nancy , Agamia Naglaa , El‐Shafaei Adel
Published: June 2021

Background

alopecia areata(AA) is a common recurrent non‐scarring type of hair loss. the pathogenesis of AA is still uncertain. The immune system has been implicated in its pathogenesis and certain immunomodulatory cytokines play a role in its development. Migration inhibitory factor(MIF) was reported to the first lymphokine which prevents the random migration of macrophages. A recent finding demonstrated that MIF innitiates inflammation and immune response by stimulating the secretion of several pro‐inflammatory cytokines. As regards skin diseases, MIF production by peripheral blood mononuclear cells has been reported to be markedly increased in patients with atopic dermatitis.

The aim of the present work was to measure the serum MIF concentration in patients with AA and in health indivduals.

Subjects and methods

The present study was carried out on 30 patients with AA and 15 healthy controls. The serum level of MIF was measured by enzyme linked immunosorbent assay (ELISA).

Results

the mean level of MIF in AA patients was 6.535ng/ml+3.51(mean+SD), while in healthy individuals was 1.89ng/ml+0.91(mean+SD)with statistical sinificant difference (p‐0.016).

Conclusion

Mean serum MIF in patients with AA was higher than that in healthy subjects indicating that MIF has a role in the etiopathogenesis of AA
The effects of corn and soybean grain dusts on the asthmatic phenotype using a murine model of fungal allergic asthma
Authors: Sharma Akshat , Hoselton Scott A , Ghosh Sumit , Schuh Jane M
Published: June 2021

Agricultural workers and farming communities are repeatedly exposed to grain dust during harvest, transport, and storage. The recognition that this complex mixture of particulates can trigger acute respiratory distress is not new, but the ways in which it may exacerbate both immediate and long‐term outcomes of allergic asthma remain unclear. The objectives of this study are to investigate the impact of repeated grain dust inhalation on the immunologic mechanisms that drive acute and chronic changes in the allergic lung. Using an inhalation model of Aspergillus fumigatus‐induced allergic asthma in mouse, we are investigating the effect of repeated exposures to corn and soybean grain dusts (collected from the rafters of commercial grain elevators). Our early data has revealed a prominent, but short‐lived, neutrophilia in the airways of naïve mice subjected to a single, 20‐min dust exposure. Continuing, we will compare airway hyperresponsiveness and inflammatory responses after multiple grain dust exposures in naïve and allergic lungs, as well as the chronic architectural changes that account for much of the morbidity of long‐term asthma.

This pilot project was funded through Central States‐Centre for Agricultural Safety and Health (CDC/NIOSH/OD).
Inhaled Carbon Monoxide Accelerates Resolution of Inflammation via Novel Pro‐resolving Mediators and Heme Oxygenase‐1
Authors: Shinohara Masakazu , Chiang Nan , Dalli Jesmond , Mirakaj Valbona , Choi Augustin M.K. , Serhan Charles N.
Published: June 2021

Resolution of acute inflammation is an active event accompanied by biosynthesis of specialized pro‐resolving mediators (SPM). We employed a systems approach to determine the impact of carbon monoxide (CO) in resolution active programs. In zymosan induced murine peritonitis model, inhaled CO gas (250 ppm) significantly limited PMN infiltration (~44% reduction at 6h) into peritoneum and shortened resolution interval. We profiled exudate lipid mediators (LM), CO reduced leukotriene (LT) B4 (21±11 vs. 59±24 pg/mouse at 6h) and elevated SPM including resolvin (Rv) E2 (1,201±197 vs. 864±125 pg/mouse), RvD1 (27±4 vs. 16±5 pg/mouse) and maresin 1 (MaR1; 26±9 vs. 15±3 pg/mouse). With human macrophages, SPM (10 pM–10 nM) elevated heme oxygenase (HO)‐1 (~50% at 8h). CO also enhanced HO‐1 expression (~25%), an action reversed by blockage of a key SPM biosynthesis enzyme 15‐lipoxygenase (LOX) type 1. CO increased phagocytosis with human macrophage, that was further enhanced by SPM. This CO increased phagocytosis was blocked by 15‐LOX inhibition and SPM stimulated phagocytosis was diminished by HO‐1 inhibition. In murine peritonitis, CO inhalation significantly increased numbers of macrophages carrying ingested apoptotic PMN at 12h in both exudates (1.9 vs. 3.4%) and inguinal lymph nodes (2.0 vs. 4.7%).

These results indicate that CO accelerates resolution of acute inflammation, shortens resolution intervals, enhances macrophage efferocytosis and temporally regulates local levels of LM/SPM. Moreover, they provide pro‐resolving mechanisms for HO‐1/CO that is part of the SPM‐initiated resolution circuit.
Concussive injury elicits human cerebrovascular endothelial cell activation in vitro
Authors: Augustine Claudia , Cepinskas Gediminas , Fraser Douglas
Published: June 2021

Concussive brain injury may directly injure cerebrovascular endothelial cells (CVEC) of the blood‐brain barrier and/or CVEC may be altered by circulating trauma‐induced inflammatory mediators. Thus, we assessed for activation of CVEC by either a concussive injury or by application of plasma from trauma patients.

Methods

Plasma was obtained from pediatric multisystem trauma patients (injury severity score≥12) and age/sex‐matched controls. Human immortalized CVEC (hCMEC/D3; provided by Dr. P. Couraud, INSERM) were employed to assess for activation in vitro. Concussive injury was induced in hCMEC/D3 grown on flexible supports with a compressed air pulse at 4 psi/well. In parallel, uninjured CVEC were treated with 20% v/v blood plasma collected from trauma patients or healthy controls.

Results

A single concussive injury to hCMEC/D3 resulted in increased nitric oxide (NO; DAF‐FM nitrosation) at 15 and 30 minutes, increased reactive oxygen species (ROS; DHR‐123 oxidation) at 2 hours and increased 51Cr‐PMN adhesion at 6 hours. In contrast, application of 20% v/v trauma patient plasma to hCMEC/D3 failed to induce oxidative stress, activate NF‐κB (ELISA) or elicit PMN adhesion assessed under “flow” (relative to control plasma‐treated hCMEC/D3).

Conclusions

Direct concussive injury induces hCMEC/D3 activation in vitro, whereas application of 20% v/v trauma plasma is not potent enough to activate CVEC.
Selective Inhibition of PKC theta Protects Murine Hearts against Ischemia/Reperfusion Injury: Cross‐talk with T Lymphocytes
Authors: Li Zhao , Abdullah Chowdhury S. , Jin Zhu‐Qiu
Published: June 2021

PKC theta is vital for activation of mature T lymphocytes. Infiltration of T cells in myocardial tissue has been observed in patients with coronary heart disease. The mechanistic role of T cells in heart injury remains elusive. In this study, effect of deactivation of mature T cells on myocardial ischemia/reperfusion (IR) injury was elucidated with two approaches: Pharmacological inhibition with a selective PKC theta inhibitor (PKCI) and genetic manipulation with Rag 1 knockout mice. Male C57BL/6J mice and Rag1 knockout mice (20 ~ 25 g) were anesthetized. Langendorff hearts perfused with Krebs‐Henseleit were prepared. Hemodynamic parameters were recorded. Creatine kinase (CK) and infarct size were measured. T cells counting, phosphorylation of PKC theta at T538 and PKC theta translocation were determined. Pretreatment with PKCI at 1 microM or 100 nM protected hearts against 50 min of global ischemia and 50 min of reperfusion as evidenced by low CK release and decreased infarct size in C57BL/6J hearts. PKCI reduced the expression of phosphorylation of PKC theta and PKC theta translocation. Myocardial IR injury was attenuated in Rag1 knockout mice, denoted by small portion of infarct size and low CK release. PKCI‐induced cardioprotection was abrogated in Rag1 knockout mice. The present results reveal that deactivation of mature T cells protects hearts against ischemia and reperfusion injury.
Prostaglandins, but not sympathetic nervous system, modulate Renin/Prorrenin receptor expression during normal pregnancy and pregnancy‐induced hypertension in rats
Authors: Lopez Pedro , Cortes‐Tinajero Viridiana , Cardenaz‐Vega Rocio , Anguiano‐Robledo Liliana
Published: June 2021

Pregnancy is associated with blunted response to contractile agents as angiotensin II (Ang II), catecholamines and prostaglandins (PG's). In contrast, their responses are increased in pregnancy‐induced hypertension (PIH). The renin‐angiotensin system (RAS) has become more complex with new components with cardiovascular functions. Renin/Prorenin receptor (RPR) is one of these components. The aim of this work was to elucidate if sympathetic nervous system (SNS) and/or PG's modulate RPR expression in normal and pregnancy‐induced hypertensive pregnant rats. We used a subrenal aorctic coarctation (SRAC) model. Using immunoblot, RT‐PCR and immunohistochemistry, we determined RPR expression in aorta and kidney from normal and PIH animals. Similar groups were treated with Prazosine (P, 2mg/Kg), Metoprolol (M, 5 mg/Kg), Indomethacine (I, 3 mg/Kg) or Celecoxib (C, 10 mg/Kg) during the third week of pregnancy. Neither P, M nor Cox inhibitors altered blood pressure. In normal pregnancy, RPR increased its expression significantly. RPR expression during PIH decreased even lower than control levels. Adrenergic blockers did not affect RPR expression significantly. I and C, in contrast, normalized RPR expression during non‐complicated pregnancy and increased it to higher levels during PIH. These results suggest that PG's, but no SNS, modulates RPR expression. Supported by SIP‐IPN 20120122 and 20120929 grants
Identification of a novel dopaminergic agonist that selectively activates the D3 dopamine receptor
Authors: Moritz Amy E. , Free R. Benjamin , Conroy Jennie , Doyle Trevor , Southall Noel , Ferrer Marc , Donthamsetti Prashant , Javitch Jonathan A. , Sibley David R.
Published: June 2021

Dopamine receptors (DARs) are involved in the development and/or treatment of many neuropsychiatric disorders, with specific receptor subtypes playing distinct roles. D2 DAR antagonism is thought to be responsible for the efficacy of antipsychotics, while D3 DAR agonists may be useful as neuroprotective and neurorestorative antiparkinsonian medications. Many antipsychotic drugs belong to the class of 1,4‐disubstituted aromatic piperazines and piperidines (1,4‐DAPS), although many produce limiting side effects due to the high conservation of the orthosteric binding site among DAR subtypes and other related GPCRs. In order to identify novel small‐molecule ligands of DARs with the potential to be highly selective and therefore better drug candidates, our lab has employed a high throughput screening (HTS) approach. Through the NIH Molecular Libraries Program, a novel 1,4 DAP, Compound 3843, was originally identified as a D2 antagonist in a D2‐Gqi5 calcium HTS of a 370,000+ small molecule library. Counter‐screening assays of beta‐arrestin recruitment revealed that this compound selectively activates the D3 DAR and acts as an antagonist at the D2 DAR. Compound 3843 also functions as a D3 DAR agonist in a Go BRET activation assay. We ultimately hope Compound 3843, or its analogs, will be a useful in vitro or in vivo pharmacological tool.
Inhibition of voltage‐gated Ca2+ channels by sst4 stimulation is mediated by Gβγ and PKC activation
Authors: Farrell Spring R , Rankin Donald R , Brecha Nicholas C , Barnes Steve
Published: June 2021

Healthcare System, Los Angeles, CA Stimulation of somatostatin subtype‐4 receptors (sst4) in isolated retinal ganglion cells (RGCs) inhibits Ca2+ channel current (ICa). Reducing intracellular Ca2+ is known to be neuroprotective; thus, stimulating sst4 may prevent RGC loss in trauma and disease. The aim of this study was to determine the signaling pathways involved in sst4 stimulation leading to suppression of ICa in RGCs. Isolated RGCs were prepared using a modified Thy‐1 immunopanning procedure. Electrophysiology experiments were performed using whole‐cell patch clamp to isolate ICa. L‐803,087 (sst4 agonist, L‐803) inhibited ICa by 40.4% and reduced calcium conductance (2.1 vs. 1.4 nS; p<0.05). Pretreatment of cells with pertussis toxin did not prevent the action of L‐803 (34.7%). Pre‐pulse facilitation did not reverse the inhibitory effects of L‐803 on ICa (11.4 vs. 9.7 %). However, pharmacologic inhibition of Gβγ prevented ICa suppression by L‐803 (23.0%, p<0.05). Inhibition of PKC (GF109203X; GFX) showed a concentration‐dependent effect in preventing the action of L‐803 on ICa (1 μM GFX, 34.3%; 5 μM GFX, 18.4%, p<0.05). This suggests that sst4 stimulation modulates RGC Ca2+ channels via Gβγ and PKC activation. Thus, modulating these signaling pathways may provide unique therapeutic targets to reduce intracellular Ca2+ levels in RGCs. Support from CIHR‐NSHRF RPP, NIH.
Depolarization of collecting lymphatic endothelium with acetylcholine or TRPV4 activation
Authors: Behringer Erik J , Scallan Joshua P , Davis Michael J , Segal Steven S
Published: June 2021

Smooth muscle cells of collecting lymphatic vessels undergo coordinated contractions to propel lymph centrally. Whether endothelial‐derived hyperpolarization [via activation of small‐ and intermediate‐conductance Ca2+ activated K+ channels (SKCa/IKCa)] shown in blood vessels is manifest in governing lymphatic function has not been established. To test this possibility, lymphangions dissected from mouse popliteal fossae were treated to yield intact lymphatic endothelial cell tubes (LECTs; width: ~50μm; length: ~500 μm) for intracellular recording (bath pH 7.4; 30°C). Resting membrane potential (Vm) of LECTs was −60±5 mV and depolarized (P<0.05) to −49±5 mV with BaCl2 (100 μM; n=6). Activation (NS309, 1 μM) or blockade [apamin (300 nM) + charybdotoxin (100 nM)] of SKCa/IKCa did not alter Vm (n=4). Acetylcholine (ACh, 1 μM) or GSK (TRPV4 channel agonist; 100 nM) depolarized LECTs by 29±7 and 49±4 mV, respectively (P<0.05; n=4 each). Thus blockade of inward‐rectifying K+ channels and agents promoting Ca2+influx (via TRPV4) as well as release from the endoplasmic reticulum (ACh) evoke LECT depolarization that is facilitated by lack of functional SKCa/IKCa expression. With myoendothelial coupling, endothelium‐derived depolarization may coordinate contraction of lymphatic smooth muscle cells and thereby determine the effectiveness of lymph transport. (NIH R37HL041026, R01HL086483, F32HL110701, R01HL089784)
Dynamics of Nitric Oxide Activity are Sufficient and Optimal to Drive Lymphatic Pumping
Authors: Kunert Christian , Padera Timothy P , Munn Lance L
Published: June 2021

The lymphatic system is critical for maintaining fluid homeostasis and immunosurveillance in tissues, but it malfunctions in many pathologies. Active pumping of fluid by collecting lymphatic vessels is a critical component of lymphatic function, transferring fluid from initial lymphatic beds, but the details of this mechanism are not well understood. Previous work has shown that lymphatic vessels are equipped with one‐way intraluminal valves to maintain unidirectional flow, and that the contraction of collecting lymphatic vessels is effected by nitric oxide (NO). Here we present a computational model that reproduces the relevant dynamics of lymphatic contraction based on known biological principles of nitric oxide combined with fluid dynamics. Shear stresses imparted by the flowing lymph induce nitric oxide locally, which cause a relaxation (dilation) of the vessel wall. The subsequent rapid degradation of NO allows for vessel contraction and fluid movement, which combined with intraluminal valves results in efficient and robust pumping.

We hypothesized that cyclic changes in shear stress and the resulting nitric oxide levels are able to produce and sustain pumping in a lymphatic vessel. To test this, we simulated lymphatic pumping using a lattice Boltzmann (LB) approach. The vessel walls consist of moving boundaries affected by a force proportional to the NO concentration and a damped, harmonic restoring potential. The NO decay and diffusion are calculated by a finite difference scheme. Our simulations show that lymphatic pumping is a stable and robust process, able to operate over a range of parameter space. Contributing to this stability is the short lifetime of NO, optimized for efficient, sustained pumping. We also demonstrate how gravity influences the pumping and that no external steering mechanism is needed.
Involvement of NO/sGC, but not ROCK, in histamine‐induced collecting lymphatic relaxation
Authors: Kurtz Kristine M , Souza‐Smith Flavia M , Breslin Jerome W
Published: June 2021

We hypothesized that histamine causes endothelial NO release, activating soluble guanylate cyclase (sGC), leading to inhibition of Rho Kinase (ROCK) to decrease lymphatic contractility. Isolated rat mesenteric collecting lymphatics displaying spontaneous contractions at a 2 cm H2O luminal pressure at 37ºC were studied before and after 10−6–10−4 M histamine treatment. Vessels were loaded with Fura‐2 to determine intracellular free calcium. The role of NO/sGC signaling was tested with 10−5 M L‐NAME or 10−4 M ODQ. The role of ROCK was investigated by transfecting vessels with constitutively active (ca)‐ROCK protein (2 μg/ml). Videos were acquired and end diastolic diameter (EDD), end systolic diameter (ESD), and contraction frequency were measured. Phasic contraction amplitude was calculated as AMP = EDD – ESD. Passive diameter (MaxD) was measured in Ca2+‐free Ringer's and used to normalize data and calculate tone: (MaxD‐EDD)/ MaxD)*100%. Histamine increased lymphatic EDD, decreased tone and AMP, and caused cessation of phasic contractions and associated Ca2+ transients. Pretreatment with L‐NAME or ODQ attenuated the response. Transfection of ca‐ROCK decreased EDD and increased tone within 5 min, but did not block histamine‐induced lymphatic relaxation. The data suggest that NO/sGC mediates histamine‐induced lymphatic relaxation, but inhibition of ROCK is not involved. Supported by NIH R01HL098215.
Effect of Acute Moderate Exercise on Vaccination Induced Inflammation and Arterial Function in Older Adults
Authors: Ranadive Sushant Mohan , Kappus Rebecca M , Yan Huimin , Lane Abbi D , Cook Marc D , Woods Jeffery A , Wilund Kenneth W , Vanar Vishwas , Fernhall Bo
Published: June 2021

Acute systemic induced inflammation negatively affects arterial function in young healthy adults. An acute bout of aerobic exercise reduces arterial stiffness and improves endothelial function. Therefore, we evaluated endothelial function and wave reflection in older adults in response to acute systemic inflammation and the effect of moderate intensity endurance exercise immediately prior to induced inflammation. 59 healthy volunteers (range 55 – 75 years) were randomly allocated to an exercise or control group. Arterial function and inflammatory markers were measured at baseline, 24 h and 48 h after influenza vaccine and sham injections. CRP increased approximately 1.5 times, 24 and 48 h after vaccination while IL‐6 increased 24 hours after vaccination compared to the sham injection. Unexpectedly, arterial function was unaltered (Table). Inflammation and endothelial function were dissociated following vaccine‐induced acute systemic inflammation in older adults. Acute exercise prior to vaccination had no effect of inflammation or vascular function. Support: AHA #10PRE3500083 (SMR).