bluuebunny/crossref_metadata_2025 huggingface dataset:
Anaemia in Infancy in Rural Bangladesh: the Contribution of Iron Deficiency (ID), Infections, and Poor Feeding Practices
Authors: Rawat Rahul , Saha Kuntal Kumar , Kennedy Andrew , Rohner Fabian , Ruel Marie T , Menon Purnima
Published: June 2021

This study aimed to determine the contribution of ID, infections and feeding practices to anaemia in Bangladeshi infants aged 6–11 months. Baseline data from 1600 infants recruited into a cluster‐randomized trial testing the effectiveness of micronutrient powder sales by frontline health workers on the prevalence of anaemia were used. Multivariate logistic regression was used to identify risk factors for anaemia and ID, and population attributable fractions (PAF) were computed to estimate the proportion of anaemia that might be prevented by the elimination of individual risk factors. Sixty‐eight percent of infants were anaemic, 56 percent iron‐deficient, and one‐third of infants had evidence of sub‐clinical infections. The prevalence of anaemia and ID increased rapidly, until 8 to 9 months of age, while the prevalence of sub‐clinical infections was constant. ID (Adjusted Odds Ratio (AOR): 2.5–2.7; p<0.001) and sub‐clinical infections (AOR: 1.4–1.5; p<0.01) were major risk factors for anaemia, in addition to age, and male sex. Sub‐clinical infections, age, and male sex, were similarly significant risk factors for ID. Previous day consumption of iron‐rich foods was very low and not associated with anaemia or ID. These results suggest a multipronged strategy is needed to prevent anaemia during infancy in Bangladesh that combines improvements in dietary iron intake alongside infection control strategies.
Characterization of anemia and iron deficiency in 6–12 month old infants from rural India
Authors: Nair K Madhavan , Fernandez‐Rao S , Balakrishna N , Radhakrishna KV , Ravinder P , Flower A Little , Hurley K M , Tilton N , Harding K , Reinhart G , Black M M
Published: June 2021

Determining anemia and iron deficiency (ID) prevalence estimates in infants using stringent criteria are a critical step in designing effective prevention strategies. The objective was to characterize biomarkers of iron, folate and vitamin B12 status and inflammatory marker C ‐ reactive protein (CRP) in 6–12 month old infants. We enrolled 497 infants from rural Andhra Pradesh, India as part of an interventional study of simultaneous early learning stimulation and at‐home micronutrient fortification. At baseline, blood was collected from 483 infants. ID was diagnosed based on ferritin <12 μg/L or serum transferrin receptor (sTfR) >;2.5 mg/L and ID with hemoglobin (Hb) <11 g/dL as iron deficiency anemia (IDA) and cases which did not fit into the above categories as indeterminant. Prevalence of anemia was 67%: iron replete infants accounted for 27%; IDA 52%; ID 6% and indeterminant cases 15%. The prevalence of folate and vitamin B12 deficiency was <1% and 19%, respectively. Mean folate status was comparable among groups while vitamin B12 status was significantly lower in IDA compared to normal infants. Indeterminant infants exhibited significantly lower Hb and sTfR and higher ferritin and CRP compared to normal infants. These findings suggest that IDA is highly prevalent among infants. Research support: Mathile Institute & Micronutrient Initiative
The predictive ability of inflammatory biomarkers in patients with newly diagnosed Inflammatory Bowel Disease [IBD]: Analyses of preliminary pre‐ and post‐therapy data
Authors: Zello Gordon , Morris Marc W , Fowler Sharyle , Alcorn Jane , Jayeoba Wemi , Rezaie Elham , Jones Jennifer
Published: June 2021

Predictors of disease course in IBD would be beneficial for early determination of patient risk and appropriate medical therapy. To evaluate their predictive performance, endoscopic appearance, fecal calprotectin (Cp) and serological high sensitivity C‐reactive protein (hsCRP), markers of inflammation, were assessed in a prospective IBD (Crohn's disease [CD] and ulcerative colitis [UC]) inception cohort (<12 mo of diagnosis). Patients were observed at baseline and subsequent clinical visits (3 or 6 mo), where detailed endoscopic, clinical disease activity, and biomarker measurements were assessed. Of the 24 patients enrolled in the study, 14 were diagnosed with CD and 10 with UC. At enrolment, the median hsCRP was 4.1 mg/L (normal 0–7.0), and Cp was 644 μg/g (<50 normal). After therapy, the median Cp was 194 and 149 ug/g in CD (n=7) and UC (n=6), respectively. When stratified by clinical disease activity, no meaningful correlation between hsCRP and Cp was apparent, in contrast to a positive correlation with endoscopic appearance. Although the sample size is small, preliminary data suggest that the use of clinical disease activity measures in newly diagnosed IBD may be inadequate for determining inflammatory burden and for risk stratification. (Support from the Royal University Hospital Saskatoon and NSERC)
A prudent diet attenuates lung function decline in cigarette smokers
Authors: Tang Wenbo , Guertin KA , Arnold KB , Hartline JA , Minasian LM , Lippman SM , Klein E , Cassano PA
Published: June 2021

Background

Most previous studies of diet and lung function focus on individual nutrients and/or foods. Dietary pattern analysis is a strong alternative approach, but very few studies examine the dietary pattern—lung function decline association.

Methods

The association between dietary patterns and decline of forced expiratory volume in 1 s (FEV1) was investigated in 2,560 male adults. Dietary patterns were derived from food frequency data using principal component analysis, and linear mixed‐effects models examined these patterns with FEV1 decline over ~3 years, adjusting for confounders. Given the importance of smoking, dietary pattern × smoking interactions were tested.

Results

A more prudent pattern (higher intake of fruits, vegetables and whole grains) was associated with a slower decline in FEV1 in smokers only (top vs. bottom tertile of pattern: +37.8 mL/y; trend Pinteraction = 0.0008). A more Western pattern (higher intake of meat, potatoes and snacks) was associated with a lower FEV1 at study baseline (top vs. bottom quintile: −162.4 mL; P for trend = 0.001), but not with FEV1 decline either overall or by smoking.

Conclusions

In males, a prudent dietary pattern was associated with a slower decline in lung function in smokers only, and a Western dietary pattern was associated with lower levels of lung function, but not change over time.

Grant Funding Source: NHLBI R01HL071022 (PAC), and, in part, NCI/DCP CA37429, NCCAM.
Relationship of maternal vitamin C intake with fetal and infant growth: Mothers and Children′s Environmental Health (MOCEH)
Authors: Kim Hyesook , Jang Won , Ha Eun‐Hee , Park Hyesook , Ha Mina , Kim Yangho , Hong Yun‐Chul , Chang Namsoo
Published: June 2021

Oxidative stress during pregnancy plays an important role in fetal growth. Vitamin C plays a major role in providing the antioxidant defense against oxygen free radicals. We examined the associations of maternal vitamin C intake with fetal and infant growth using data from 1216 participants in the Mothers and Children's Environmental Health (MOCEH), a multi‐center prospective cohort study in South Korea. Dietary intakes in mid pregnancy were estimated by a 24‐hour recall method, and infant weights and heights were measured at birth and 6 months after birth (n = 1216 and 933, respectively). Multiple regression analysis showed that maternal vitamin C intake was positively associated with birth height (P = 0.0513) after adjustment for covariates that may affect fetal growth. We also observed a positive association between vitamin C intake and height at 6 months after birth (P = 0.0222). Results of this study suggest that higher intake of vitamin C in mid pregnancy is associated with higher infant height at birth and 6 months after birth.
Fathers’ acceptance of videoconferencing for remote lactation consultation: a pilot study
Authors: Lewis Daniel Gregory , Habibi Mona F , Kavanagh Katherine F
Published: June 2021

The objective was to pilot a survey designed to assess fathers’ perceived acceptability of videoconferencing technology for the purpose of remote lactation consultation (RLC) for their partners. Data were obtained via a voluntary web‐based survey advertised on social media websites. Fathers >; 18 years old, with an infant < 6 months of age who was receiving breast milk at the time of study participation were eligible. Forty‐one fathers were eligible, consented, and completed the survey. Fathers were asked about their comfort with their partners’ use of web‐based communication software, general comfort levels with their partners’ breastfeeding, about any breastfeeding problems encountered, and any subsequent actions taken to correct these problems. Comfort with using web‐based communication software was generally high, with 81% (n=33) of fathers reporting comfort with their partners using email or instant messaging to communicate with health professionals, and 84% (n=35) reporting that they would feel comfortable with partners using web‐cams to communicate with health professionals. Of those reporting partners experiencing breastfeeding problems, 74% (n=30) of these problems would be included within the scope of practice of an International Board Certified Lactation Consultant. Results of this pilot study indicate acceptance among fathers of videoconferencing for the purpose of RLC.

Grant Funding Source: University of Tennessee Internal Seed Funds
The role of protein O‐mannosylation during mouse development
Authors: Strahl Sabine , Lommel Mark , Willer Tobias , Schneider Marlon R , Dahlhoff Maik , Wolf Eckhard
Published: June 2021

Protein O‐mannosylation is an essential modification among fungi and mammals. It is initiated at the endoplasmic reticulum by a conserved family of dolichyl phosphate‐mannose:protein O‐mannosyltransferases (PMT/POMTs). Mutations in POMTs cause dystroglycanopathies covering a wide spectrum of clinical severities ranging from Walker‐Warburg syndrome to limb‐girdle muscular dystrophy. In mammals, the role of O‐mannosyl glycans in the binding of α‐dystroglycan to components of the extracellular matrix such as laminin is well defined. Other functions of O‐mannosyl glycans, however, are poorly characterized. Here, we will discuss our recent data demonstrating a role of O‐mannosyl glycans during embryo development that is largely independent of α‐dystroglycan.
Estrogen replacement prevents ovariectomy‐induced muscle degradation via lowering local IGF‐1 production
Authors: Ezzat‐Zadeh Zahra , Navaei Negin , Akhavan Neda , Park Bong‐Sup , Chase P. Bryant , Kim Jeong‐Su , Arjmandi Bahram H.
Published: June 2021

The prevalence of sarcopenia in women increases following menopause. Although muscle mass and circulating estrogens are positively correlated, the underlying mechanisms by which estrogen deficiency causes muscle atrophy is poorly understood. The present study examined the role of systemic and local IGF‐I in muscle maintenance of estrogen deficient rats. Eighteen 10‐month old female SD rats (n=6/group) were divided into either a sham operated group (sham) or two ovariectomized (ovx) groups receiving 17β‐estradiol (ovx+E2) or solvent vehicle (sham and ovx). All animals were pair‐fed for five months. Ovarian hormone deficiency did not cause atrophy in gastrocnemius (GAS) and soleus (SOL) muscle; however when muscle weight was normalized to body weight, the mean GAS value of ovx group was significantly lower than those of sham and E2 groups. Similarly, ovx mean SOL value tended (P<0.094) to be lower compared with other two groups. E2 treatment was able to prevent the ovx‐induced decreases in SOL, and GAS normalized values by 50% and 100%, respectively. Ovx significantly (P<0.05) increased systemic IGF‐I by 38% while local IGF‐I mRNA expressions were significantly decreased in both GAS (29%) and SOL (43%) samples of ovx rats. E2 administration prevented these decreases in IGF‐I levels. Our findings suggest that ovx‐induced elevation of systemic IGF‐I may down‐regulate its local mRNA expressions which in part can explain the loss of muscle mass in ovarian hormone deficiency.

Grant Funding Source: Internal Funding
DHA dampens endocannabinoid tone related to cannabinoid receptor expression in proliferating C2C12 myoblasts
Authors: Kim Jeffrey , Li Yong , Alzghoul Borhan , Watkins Bruce A
Published: June 2021

Emerging evidence indicates that the peripheral endocannabinoid signaling system (ECS) has a pronounced effect on health status, specifically glucose homeostasis. Focus on skeletal muscle is of great interest as it is a major storage for glycogen in the body. However, overactivation of the ECS in skeletal muscle has been shown to diminish insulin sensitivity. In particular, an elevation of CB1 activity has been reported in diabetic rodent models. Our research intends to identify changes in ECS tone by fatty acid (FA) enrichment. Proliferating C2C12 myoblasts were cultured until 50% confluency and treated with either 25 μM of AA, EPA, AEA, 2‐AG, or a BSA control for 24 h. Cells were collected and labeled with antibodies for CB1 and CB2 for FACS flow cytometry and FA analysis by GC. Compared to the control, FACS revealed a significant reduction in CB1 abundance with DHA treatment, while a significantly higher expression of CB1 was found with AA. No difference in CB2 level was observed. FA composition of myoblasts reflected their respective FA treatment. Interestingly, between the two AA‐derived ECS ligands, only 2‐AG enrichment had a doubling of its precursor, AA, but not with AEA. These findings suggest that DHA may dampen an overactive EC tone.

Grant Funding Source: Diet and Health Initiative, UConn, Storrs, CT, USA
Recommended Dairy Intake: Impact on Lipid Profile and Surrogate Markers of Cholesterol Homeostasis in Healthy Adults from Two Canadian Cities
Authors: Abdullah Mohammad , Cyr Audrey , Couture Patrick , Jones Peter , Lamarche Benoît
Published: June 2021

We assessed the impact of dairy intake on lipid profile and surrogate markers of cholesterol absorption, synthesis, and clearance in a free‐living, randomized, multicentre crossover trial of two 4‐wk dietary phases, DAIY and CONTROL, separated by a 4–8 wk washout, in Winnipeg and Quebec City, Canada. As part of a healthy‐style eating pattern, participants (n= 126) consumed 3 servings/d of conventional dairy products (375 ml/d low‐fat milk, 175 g/d low‐fat yogurt and 30 g/d regular cheddar cheese) or energy‐equivalent control products (290 ml/d fruit juice, 156 ml/d vegetable juice, 20 g/d cashews and 1 cookie). The DAIRY diet had no significant impact on circulating lipids, except for low density lipoprotein (LDL) levels, which were slightly higher relative to the CONTROL diet (2.98 vs. 2.90 mmol/L, P=0.03). This was accompanied by a reduction in plasma phytosterols’ levels (−11.0%, P<0.001), with no changes in lathosterol and proprotein convertase subtilisin/kexin‐9 (PCSK9) concentrations. These data indicate that the short‐term intake of DAIRY diet had possibly lowered cholesterol absorption, but not synthesis or clearance, in comparison to the CONTROL diet. Although plasma LDL levels were raised subsequent to the dairy consumption, the variability in the response of LDL (range: −1.29 to 1.03 mmol/L) across individuals suggested a genetic determinant. Supported by the Dairy Farmers of Canada.

Grant Funding Source: Dairy Farmers of Canada
Toxicological and Cancer‐Chemopreventive Evaluation of a Novel Soluble Silicate
Authors: Vattem Dhiraj Anil , Maitin Vatsala , Richardson Charles Reed
Published: June 2021

Silicates are ubiquitous in nature and exhibit diverse speciation and unique physiochemical properties. Though abundant in diet, their biological role in animals is not clearly understood. In our previous study, we showed that a unique soluble species of silicate [Na8.2Si4.4H9.7O17.6] modulated stress response signaling pathways in vivo and may have implications in human health. Here we present results from toxicological and cancer chemopreventive evaluation of Na8.2Si4.4H9.7O17.6. Results suggest LD50 >;5000 mg/kg for acute oral toxicity of silicates in Sprague‐Dawley rats. Undiluted silicate was found to be non‐corrosive in rabbit skin corrosion study. A statistically significant reduction in fibroblast, platelet‐derived and vascular endothelial growth factor induced angiogenesis was observed in the chick embryo chorioallantoic membrane model at >;18.8 nM. A significant reduction in the size of the NM2C5 tumors was observed in vitro in the three‐dimensional tumor microenvironment model at a dosage of >;18.8 nM. At a dosage of >; 7.5 mM the silicate also decreased the tumor development and metastasis of LOX‐GFP human melanoma in Male NCr nude mice by ~15%. Our results suggest that soluble silicates may have potential cancer‐chemopreventive effects and merits further detailed investigation to understand mechanism of action.
High‐dose (4000 IU) Vitamin D Supplementation Does Not Increase Circulating Concentrations of Free 25‐hydroxyvitamin D
Authors: Belenchia Anthony M , Tosh Aneesh , Hillman Laura , Peterson Catherine A
Published: June 2021

Free/bioavailable circulating 25‐hydroxyvitamin D (25OHD) makes up a small fraction of total 25OHD concentration. We demonstrated the efficacy of a daily 4000 IU vit D dosage in raising total 25OHD concentrations of obese adolescents. However, it remained unclear whether the increase in total 25OHD was due to increases in the free form or that bound to vit D binding protein (DBP). The ratio of total 25OHD:DBP has been used as a proxy measure of free 25OHD. 44 patients (age=13.9; BMI =39.2) were recruited from the University of Missouri‐ADOBE clinic and were randomized to receive either placebo or vit D3 (4000 IU/day) treatment for 6 months. At baseline (BL), there was no difference between the vit D and placebo group in the ratio of 25OHD:DBP (0.19 vs 0.23; P=0.28). There no difference between groups in the change from BL (0.11 vs 0.05; P=0.10). There was a significant gender difference in the ratio at BL (M=0.17 vs F=0.12; P=0.015). Change from BL of total 25OHD was positively correlated with the change in the ratio (r=0.57; P<0.001) but not DBP(r=0.28; P=0.14). Supplementing obese adolescents with vit D3 does not increase free circulating 25OHD.

Grant Funding Source: J.R. Albert Foundation, inc
Trends in Mineral Supplement Prescriptions from Military Treatment Facilities: 2007 to 2011
Authors: Attipoe Selasi , Jones Donnamaria , Stephens Mark B. , Deuster Patricia A.
Published: June 2021

INTRODUCTION

Mineral supplements (MS), such as calcium and iron, are readily available over the counter and are some of the most frequently consumed dietary supplements (DS). Healthcare providers also prescribe MS for treatment of certain conditions and maintenance of health. This study examines trends in MS prescriptions at Military Treatment Facilities (MTF).

METHODS

Data from the Department of Defense (DoD) Pharmacoeconomic Center (PEC) was used to determine all MS prescriptions disbursed by MTF pharmacies between 2007 and 2011.

RESULTS

Overall, 1,797,982 calcium; 847,370 iron; 166,318 magnesium; and 23,297 zinc prescriptions were dispensed over this five‐year period. Whereas the number of zinc prescriptions decreased considerably by an average of 30% across the five‐year period, calcium and magnesium prescriptions increased slightly by 4% and 9%, respectively. Iron prescriptions dispensed were relatively stable across the five years.

CONCLUSION

Patterns of MS prescriptions change over time. Whether the patterns within the DoD medical system are comparable to the civilian medical system is unknown. Also, it is not known whether the DS being prescribed are effective for the intended use.

FUNDING:

This study was supported by Center Alliance for Dietary Supplement Research (NA91FD).
Anti‐inflammatory effect of Spirluina platensis in macrophages is beneficial for adipocyte differentiation and maturation by inhibiting NF‐κB pathway in 3T3‐L1 adipocytes
Authors: Pham Tho Xuan , Lee Jiyoung
Published: June 2021

We showed organic extract of Spirulina platensis, a blue‐green alga, (SPE) had potent anti‐inflammatory effects in macrophages. We hypothesize that by cross‐talking with adipocytes, the anti‐inflammatory effects of SPE in macrophages can be beneficial for adipocyte differentiation/maturation by inhibiting pro‐inflammatory cytokine production. Ten % conditioned medium from LPS‐stimulated RAW 264.7 macrophages (CMC) and LPS‐stimulated but SPE‐treated macrophages (CMS) were treated on 3T3‐L1 preadipocytes at different stages of differentiation. Expression of adipocyte differentiation markers, such as CCAAT/enhancer‐binding protein α, peroxisome proliferator‐activated receptor γ, and perilipin, was significantly repressed by CMC but attenuated by CMS. Oil Red O staining confirmed adipocyte maturation in CMS‐treated cells, but not CMC‐treated cells, were equivalent to control cells. Nuclear translocation of nuclear factor κB was attenuated by CMS compared to CMC. In lipid‐laden adipocytes, CMC promoted the loss of lipid droplets, while CMS had minimal effects. Histone deacetylase 3 and 9 are down‐regulated during the adipogensis, but CMS treatment reduced their abundance more than control. Our results show that SPE can promote adipocyte differentiation/maturation in inflammatory conditions, which otherwise can be compromised as in several metabolic disorders.

Grant Funding Source: NIH R21AT005152
Anti‐diabetic effect and antioxidant activity of the roasted Gugija (Lucii fructus)
Authors: Lee Kyongae , Park Jinsook
Published: June 2021

This study investigated the chemical characteristics (proximate compositions, free amino acid and mineral composition analyses), anti‐diabetic effect and antioxidant activity of its hot water‐ and ethanol‐extracts. Contents of carbohydrate, protein, lipid, and ash of the roasted Gugija were 56.3, 13.8, 12.5 and 5.1%, respectively. Three major free amino acids of the hot water‐extract of the roasted Gugija were aspartic acid (244.4 mg%), alanine (132.1 mg%), and arginine (114.7 mg%), whereas those of the ethanol‐extract were aspartic acid (174.7 mg%), alanine (10.0 mg%), and methionine (85.7mg%). The most abundant mineral in the hot water‐ and ethanol‐extracts of the roasted Gugija was potassium at 962.2 and 1023.0 mg%, respectively. Higherα‐glucosidase activity observed in the ethanol‐extract, when compared to the hot water‐extract. DPPH and ABTS radical scavenging activities of hot water‐extract were 85.1 and 80.3%, respectively, whereas those of ethanol‐extract were 88.5%, and 83.1%, respectively. The ethanol‐extract showed higher DPPH and ABTS radical scavenging activities than the hot water‐extract. Higher antioxidant activity of the ethanol‐extract than that of the hot water‐extract may be related with free amino acid compositions as well as higher polyphenol content in the ethanol‐extract (14.9 mg/g) compared to that in the hot water‐extract (12.7 mg/g).
Mango polyphenols reduce inflammation in MDA‐MB231 breast cancer cell and targets MicroRNA‐21
Authors: Arbizu Shirley , Krenek Kimberly , Mertens‐Talcott Susanne
Published: June 2021

Mango polyphenols have received some attention based on their antioxidant, anticarcinogenic and anti‐inflammatory properties. Our objective was to determine the anti‐inflammatory and cytotoxic properties of mango polyphenols (MP) in MCF12A non‐cancer breast fibroblasts and MDA‐MB231 breast cancer cells. mRNA, miRNA and protein expression of MDA‐MB231 and MCF12A cells were analyzed by real time PCR and Western blotting.

Results show that MP (5mg of gallic acid equivalents/L (GAE/L)) decreased the proliferation of MB‐231 breast cancer cells by 90%; but not of MCF‐12A cells. In MB‐231 breast cancer cells, MP downregulated the expression of NF‐□B (to 0.6‐fold of the control), VEGF (0.5‐fold), TNF‐α (0.8‐fold), survivin (0.5‐fold) and Bcl2 (0.5‐fold). In MCF‐12A breast cells, MP downregulated TNF‐α‐induced expression of IL‐6 (0.8‐fold), IL‐1β (0.6‐fold) and NFKb (0.4‐ fold). miRNA 21 is up‐regulated in breast cancer and targets Bcl2 which is involved in apoptosis. MP reduced the expression the miR‐21 in a dose dependent manner and these findings were confirmed using the specific antagomir for miR‐21, where it reversed the effect of the polyphenolic treatment in MB‐231 cells.

These results indicate that the cytotoxic effects of mango polyphenols are specific to cancer cells where inflammation was reduced in both, cancer and non‐cancer cells, seemingly through the involvement of miR‐21.

Grant Funding Source: Research Assistanship
Anti‐inflammatory activity and metabolism of α‐mangostin by cells of human origin
Authors: Gutierrez‐orozco Fabiola , Chitchumroonchokchai Chureeporn , Lesinski Gregory B , Failla Mark L
Published: June 2021

Xanthones in mangosteen (Garcinia mangostana), a fruit native to Southeast Asia, have been reported to exhibit anti‐proliferative and pro‐apoptotic activities. Recent data from our laboratory suggest ingested xanthones and their metabolites are absorbed and delivered to tissues where they may be accumulated and further metabolized. Neither the anti‐inflammatory activity of xanthones for cells of human origin nor the metabolism of these compounds by animal or human cells have not been examined with the exception of our previous studies with Caco‐2 human intestinal cells. We have found that ≤15 μM α‐mangostin (α‐MG), the most abundant xanthone in mangosteen, suppresses secretion of a) IL‐8 by activated human Caco‐2 intestinal, HT‐29 colonic, and THP‐1 macrophage‐like cell lines, and b) TNFα by activated HepG2 and human blood monocyte‐derived macrophages. α‐MG was taken up and metabolized by the various cell types and results with Caco‐2 suggest bioavailability may be enhanced during inflammatory conditions. Furthermore, unconjugated xanthones were predominant in HepG2, THP‐1 and HT‐29 cultures in the pro‐inflammatory environment. In addition to glucuronidation/sulfation of α‐MG, conversion to 9‐hydroxycalabaxanthone, garcinones C and D, and other unknown xanthone compounds was observed. Ongoing studies are examining anti‐inflammatory activity of α‐MG using a preclinical model.

Grant Funding Source: OSU FIC and Conacyt Fellowship (Mexico)
Proteomics of early azoxymethane‐induced intestinal tumorigenesis in F344 rats consuming a normal diet.
Authors: Swain James , Chance Mark , Zhao Hong , Wang Li , Yohannes Elizabeth
Published: June 2021

Aberrant Crypt Foci (ACF) are putative preneoplastic lesions, representing the earliest morphological lesion visible in colon cancer. The purpose of this study was to determine changes in the protein profile of ACF as they transform into adenomas under normal dietary conditions. Protein expression profiles of azoxymethane (AOM)‐induced F344 rat colon ACF were compared at 4, 8, 16, and 24 wk post AOM administration; time points correlate to induction and transformation events. At each time point, microdissected ACF/adenoma and adjacent normal tissue protein samples from four animals were analyzed together across multiple quantitative two‐dimensional (2D‐DIGE) gels using a Cy‐dye(Cy3, Cy5)/dye‐swap technique and a pooled internal standard sample to quantify expression changes with statistical confidence. Principle component analysis, Western blot, and subsequent network pathway mapping were used to identify/confirm differentially expressed proteins of interest, which included changes in vinculin (Vcl; p=0.007), ornithine aminotransferase (Ota; p=0.001), and stress‐induced phosphoprotein 1 (Stip1; p=0.003) in ACF and adenomas. By determining proteomic changes that occur in ACF as they transform into adenomas, key proteins related to adenocarcinoma development were elucidated. Support provided by Clinical and Translational Science Collaborative (CTSC) Core, CWRU.
Innate immune sensing of a bacterial protein in the ER by IRE1α
Authors: Cho Jin Ah , Lencer Wayne
Published: June 2021

The plasma membrane and all membrane‐bound organelles except for the Golgi and endoplasmic reticulum (ER) are equipped with pattern‐recognition molecules to sense microbes or their products and induce innate immunity for host defense. Here, we report that Inositol‐REquiring‐1α (IRE1α), an ER protein that signals in the unfolded protein response (UPR), is activated to induce inflammation by cholera and Shiga toxins as they co‐opt the ER to cause disease. Other known UPR transducers, including the IRE1α effector XBP1 are dispensable for this signaling. The inflammatory response depends instead on the RNase activity of IRE1α and retinoic‐acid inducible gene 1 (RIG‐I), a cytosolic sensor of RNA fragments. These observations implicate Regulated IRE1α‐Dependent Decay of mRNA (RIDD) in a novel innate immune mechanism of host defense originating from within the ER lumen.

Grant Funding Source: NIH R01/CCFA
Ω‐3 Fatty Acid Effects on Obese Leptin Knock‐Out Mice
Authors: Ali Arfaa , Nachnani Jagdish , Bulchandani Deepti , Alba Laura , Mansour Ahmad , Hamdan Hana , Quinn Tim , Molteni Agostino , Herndon Betty
Published: June 2021

Liver steatosis in wild‐type rats on methionine‐choline deficient diets is decreased with fish oil (Nachnani, Diabetologia 53:153,2010). Effects of Ω‐3 fatty acid were studied here in OB/OB (leptin k/o) mice with natural obesity and steatosis. Ten mice received purified Ω‐3 (20% EPA, 50% DHA) in food pellets, equivalent (mg/kg) to a human dose of ~4 grams/day. Ten more OB/OB mice received corn oil in their food. At 60 day necropsy, livers were harvested for study. Five Ω‐3 treated mice survived and unexpectedly blood glucose of >;600 mg/dL was found in 3 of them. Since leptin receptor k/o mice (DB/DB) are diabetes models, liver homogenate and sera of Ω‐3 and controls were analyzed for murine leptin receptor (ELISA), but no difference was found between groups. Ω‐3 treated mice showed reduced fatty liver infiltration, 27.6% Ω‐3 and 38.5% untreated, with less periportal and portal triad inflammation in Ω‐3 mice. Heart histopathology showed irregular, eosinophilic myocardial fibers with pyknotic nuclei, suggesting necrosis. Such changes in heart cell morphology is reported in human diabetes. Sera showed significant lowering of insulin, IL‐6, adiponectin, and TNFα in the Ω‐3 group compared to untreated. Ω‐3 fatty acid treatment may reduce liver steatosis in OB/OB mice but other physiological changes with diabetes induction may limit its use as a model.

Funded: St. Luke's Foundation, Kansas City
Inflammatory cells and cytokines in the Olfactory Bulb of a rat model of neuroinflammation; Insights into neurodegeneration?
Authors: Doursout Marie‐Francoise , Schurdell Michael S. , Young Lauren M. , Osuagwu Uzondu , Hook Diana M. , Poindexter Brian J. , Schiess Mya C. , Bick Diane L. , Bick Roger J.
Published: June 2021

This study examined inflammatory cell and cytokine production in brain tissue from a lipopolysaccharide (LPS) treated rat model that mimics many of the neuropathologic changes associated with neurodegenerative diseases. We monitored the appearance of glial cell line‐derived neurotrophic factor (GDNF) and circulating nitric oxide (NO) levels, as well as immune system associated cells in a selected area of the brain, the olfactory lobe. Our hypothesis was that LPS treatment stimulates temporal changes within the brain and these responses included immune cell recruitment, increased tissue levels of immune modulating cytokines and NO, as well as greater glial cell activation resulting in increased production of GDNF. Our animal model of systemic LPS treatment lead to an increase in the concentrations of circulating cytokines including TNF‐α, IL‐Iβ and IL‐6, with a maximum response 6 hours post LPS. Concomitant with cytokine elevations, circulating NO levels were elevated for several hours post LPS. Brain content of GDNF was also elevated over a similar time frame. Lymphocytes, neutrophils, macrophages, plasma cells and cytokines were all seen in various areas of LPS treated brains. Our results suggest an involvement of both the peripheral and the central nervous system immune components in response to inflammation and inflammatory episodes.
Silencing of drpr leads to muscle and brain degeneration in adult Drosophila
Authors: Draper Isabelle , Mahoney Lane J. , Mitsuhashi Satomi , Freeman Marc R. , Salomon Robert N. , Kang Peter B.
Published: June 2021

Mutations in MEGF10 underlie a novel autosomal recessive congenital muscle disease in humans. To explore the role of MEGF10 in muscle, we studied its Drosophila homolog, drpr, in muscle function and anatomy. We show that adult drpr null flies display shortened lifespan and locomotor defects associated with abnormal positioning of the legs. RNAi‐mediated down‐regulation of drpr expression in the entire organism, or targeted to muscle cell progenitors, results in an age‐dependent decrease in motor activity. Histological analysis of drpr mutants reveals abnormalities in both the brain and muscle, in contrast to the human phenotype which only affects skeletal muscle. The absence of functional redundancy for drpr in Drosophila may help elucidate whether paralogs of MEGF10 in humans (e.g. MEGF11) contribute to maintaining wild‐type function in the human brain. Homologous mammalian proteins may be introduced in the fly mutant background to study the in vivo function of the wild‐type protein and the effects of mutations. Our Drosophila models will help delineate the physiologic relevance of the MEGF10/drpr pathway in muscle function and disease. This study was funded by the Muscular Dystrophy Association and a Boston Children's Hospital Pilot Grant.
CAP37: A Marker of Neuroinflammation
Authors: Brock Amanda J , Kasus‐Jacobi Anne , Lerner Megan , Adesina Adekunle M , Pereira Anne
Published: June 2021

As the pathology of many neurodegenerative diseases begins several years before symptoms manifest, there is an urgent need to determine early biomarkers so proper therapies can be implemented to halt or slow disease progression. Previous findings demonstrated cationic antimicrobial protein of m.w. 37kDa (CAP37), an innate immune system molecule, to be a potent modulator of microglial functions. The objective of this study was to investigate whether CAP37 could serve as an early biomarker of neuroinflammation. Tissues from brains of Alzheimer's disease (AD), Pick's disease (PiD), and Vascular Dementia patients, and from retinas of Age‐related Macular Degeneration (AMD) patients and age‐matched controls were analyzed for the presence of CAP37 using immunohistochemistry. Our results show that CAP37 is expressed in cortical neurons in temporal and parietal lobes of AD and PiD patients, and expressed at a lower frequency in controls. CAP37 was also found in CA3 and CA4 regions of the hippocampus. In Vascular Dementia patients, CAP37 was confined to endothelial cells and neutrophils. Staining also demonstrated CAP37 in photoreceptors, endothelial cells, and drusen of AMD patients, and was absent in these regions in controls. In conclusion, based on the presence of CAP37 in key neurodegenerative diseases, our hypothesis is that CAP37 is a potential early biomarker. Support: OCAST HR12–068 & NEI 5R01EY015534
Equine Navicular Syndrome: Historical, Pathological and Biomechanical Insights into Pathogenesis and Treatment
Authors: Uhl Elizabeth W , Cornille Jean Luc
Published: June 2021

Because horses were critical for transportation and the military, equine diseases have been studied for centuries. Navicular syndrome (NS), a common cause of lameness in modern horses, was definitively described in 1816 and rapidly increased in incidence. Historical records indicate this time frame coincides with the development of macadam pavement. NS was immediately recognized as a disease of ‘work’, yet most studies have exclusively focused on the lesions in the foot. For this study the pathology of NS was reviewed and found to be consistent with compression–induced injury, which implicates overloading by aberrant biomechanical forces as a potential cause. Assessment of 4 cases of clinical NS revealed all had significant transversal rotation of the vertebral column, associated with the metacarpal bone passing the vertical before the commencement of the propulsive stage of the stride, a kinematic that was confirmed to compress the navicular structures on dissected specimens. Correction of these biomechanical abnormalities through targeted training exercises restored soundness and resulted in all horses returning to work and remaining sound. Incorporation of historical and biomechanical insights with the pathology of equine degenerative joint diseases can provide the broad perspective needed both understand and treat these devastating conditions.
Statins Inhibit NADPH Oxidase Activity by Interference with Membrane Raft Clustering Independent of Rac1 Inactivation in Endothelial Cells
Authors: Zhang Yang , Li Xiang , Li Pin‐Lan
Published: June 2021

Statins, the inhibitor of 3‐hydroxy‐3‐methylglutaryl coenzyme A (HMG‐CoA) reductase, inhibit mevalonate pathway to block cholesterol biosynthesis, thereby lowering plasma cholesterol and being used for prevention and treatment of atherosclerotic diseases. Our previous studies have shown that statins including pravastatin and simvastatin interfere with the formation of OxLDL‐induced membrane raft (MR)‐redox signaling platforms in human coronary arterial endothelial cells (HCAECs) and in mouse coronary arterial endothelium. However, it remains unknown how statins produce this action that is independent of plasma cholesterol lowering. Here, we tested a hypothesis that inhibition of enhanced MR clustering is essential for the inhibitory effect of statins on Rac1 and NADPH oxidase (NOX) activity. We first demonstrated that statins had no effect on protein expression of NOX subunits, gp91phox or p47phox and Rac1 in HCAECs and mRNA levels of gp91phox or p47phox in coronary arteries of mice, confirming that statins primarily inhibit NADPH oxidase activation rather than alter their gene expression. Overexpression of oncogenic Rac1 in HCAECs was unable to reverse the inhibitory effects of statins on OxLDL‐induced MR clustering suggesting that the effect of statins on MR clustering does not require Rac1 activation. Electron spin resonance spectrometry demonstrated that incubation of HCAECs with sphingomyelinase, a strong stimulator of MR clustering significantly increased O2•− production, which was abolished by pravastatin and simvastatin or by a MR disruptor methyl‐β‐cyclodextrin. Taken together, these results indicate that statins inhibit NOX activity by interference with MR clustering independent of Rac1 activation. This action of statins on MR clustering may be associated with reduction of cholesterol in cell membrane of ECs (supported by NIH grants HL057244, HL091464 and HL075316).
Inducible Endothelium‐Derived Hyperpolarizing Factor (iEDHF): A New Role for the Endothelial 15‐Lipoxygenase Pathway
Authors: Campbell William B , Gauthier Kathryn M , Pfister Sandra L
Published: June 2021

Endothelium‐derived hyperpolarizing factors (EDHFs) mediate a portion of the vasorelaxations to agonists such as acetylcholine (ACh). In rabbit aorta and mesenteric arteries, ACh and arachidonic acid (AA) caused endothelium‐dependent relaxations that were inhibited by lipoxygenase (LO) inhibitors. Arteries metabolized AA to 15(S)‐hydroxy‐11,12‐epoxyeicosatrienoic acid (HEETA) and 11(R),12(S),15(S)‐trihydroxyeicosatrienoic acid (THETA). LO inhibitors reduced their synthesis. HEETA and THETA are EDHFs that activate smooth muscle cell 24 pS, calcium‐activated potassium channels causing membrane hyperpolarization and relaxation. Endothelial 15‐LO expression was increased by adenoviral transduction of arteries and resulted in increased synthesis of HEETA and THETA and increased relaxations to AA and ACh. Hypoxia, hypercholesterolemia, estrogen and interleukin‐13 increased the endothelial expression of 15‐LO. These increases in endothelial 15‐LO expression enhanced HEETA and THETA synthesis, increased membrane hyperpolarization and increased relaxations to ACh and AA. 15‐ LO is an inducible enzyme and changes in its expression regulate EDHF activity. Thus, the 15‐LO/HEETA/THETA pathway represents the first example of an inducible EDHF. (Supported by HL‐37981, HL‐93181 and HL‐103673)
A‐kinase anchoring proteins (AKAPs) regulate airway smooth muscle secretory function
Authors: Poppinga Wilfred J , Oldenburger Anouk , Holtzer Laura J , Driessen Saskia , Heijink Irene H , Maarsingh Harm , Schmidt Martina
Published: June 2021

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory disease mainly caused by cigarette smoke. Current pharmacotherapy involves β2‐agonists and phosphodiesterase inhibitors. Both drugs elevate cAMP, but relieve a distinct subset of COPD symptoms. This may be mediated by A‐kinase anchoring proteins (AKAPs) that compartmentalize cAMP signaling. Here we studied the role of AKAPs in human airway smooth muscle (ASM) secretory function. Using western blot, radio‐assays for PKA binding (RII overlay) and RT‐qPCR we show that human ASM cells express AKAP5, AKAP8, AKAP9, AKAP12 and ezrin, and that cigarette smoke extract (CSE) decreases AKAP5, AKAP9 and AKAP12 expression. In line, we observed reduced mRNA expression of these AKAPs in lung tissue homogenates of COPD patients compared to healthy controls. Disruption of AKAP functions using st‐Ht31 augmented CSE‐induced IL‐8 release and prevented the inhibitory effect of the β2‐agonist fenoterol on CSE‐induced IL‐8 release. St‐Ht31 also diminished the inhibitory effect of fenoterol on CSE‐induced ERK phosphorylation, but not NFκB activation. Thus, the anti‐inflammatory effect of fenoterol on ASM secretory function is mediated by AKAPs. The reduction of specific AKAPs in COPD may disrupt compartmentalized cAMP and thereby affect pharmacotherapy. Supported by Dutch Asthma Foundation grant 3.2.11.015 and a Rosalind Franklin Fellowship.
Adenosine receptor subtype 3 (A3AR) agonists as novel analgesics in chronic neuropathic pain
Authors: Salvemini Daniela , Janes Kali , Finley Amanda , Doyle Timothy , Chen Zhoumou , Bryant Leesa , Tosh Dilip , Jacobson Kenneth
Published: June 2021

Chronic neuropathic pain is poorly managed and represents a huge unmet medical need. In chronic constriction injury (CCI) and chemotherapy‐induced neuropathic pain (CIPN), we demonstrate the analgesic effects of potent and specific A3AR agonists, IBMECA or Cl‐IBMECA; now in clinical trials as anti‐inflammatory and antitumor agents. In mice and rats with CCI, systemic A3AR agonists reversed mechano‐allodynia in a dose‐dependent manner. These effects were blocked by a selective A3AR antagonist (MRS1523), lost in A3AR−/− mice, and had both peripheral and spinal sites of action that were attenuated by intraplantar or intrathechal injection of MRS1523. CIPN is the major toxicity of widely used antitumoral agents and one of the most common causes of dose reduction and discontinuation of an otherwise life‐saving therapy. In rats, IB‐MECA and Cl‐IB‐MECA blocked or reversed mechano‐allodynia and mechano‐hyperalgesia induced by paclitaxel, oxaliplatin and bortezomib in a dose‐dependent manner without interfering with antitumor effects. The selective and oral A3AR agonists, MRS1898 and MRS5698 were equally effective. Our findings provide the pharmacological rationale towards the clinical development of A3AR agonists for chronic pain of diverse etiologies. Supported by NIH/NIDA [R01 DA024074], NIH/NIHMS [5T32GM008306], Saint Louis University President Research Funds, St. Louis Cancer Center.
Cannabidiol protects against hepatic ischemia/reperfusion injury by attenuating inflammatory signaling and response, oxidative/nitrative stress, and cell death
Authors: PACHER PAL , Mukhopadhyay Partha , Rajesh Mohanraj , Horvath Bela , Batkai Sandor , Park Ogyi , Haskó György , Liaudet Lucas , Wink David , Mechoulam Raphael
Published: June 2021

Ischemia/reperfusion (I/R) is a pivotal mechanism of liver damage after liver transplantation or hepatic surgery. We have investigated the effects of cannabidiol (CBD), the nonpsychotropic constituent of marijuana, in a mouse model of hepatic I/R injury. I/R triggered time‐dependent increases/changes in markers of liver injury, hepatic oxidative/nitrative stress, mitochondrial dysfunction, inflammation, stress signaling, and cell death. CBD significantly reduced the extent of liver inflammation, oxidative/nitrative stress, and cell death and also attenuated the bacterial endotoxin‐triggered NF‐κB activation and TNF‐α production in isolated Kupffer cells, likewise the adhesion molecule expression in primary human liver sinusoidal endothelial cells stimulated with TNF‐α and attachment of human neutrophils to the activated endothelium. These protective effects were preserved in CB2 knockout mice and were not prevented by CB1/2 antagonists in vitro. Thus, CBD may represent a novel, protective strategy against I/R injury by attenuating key inflammatory pathways and oxidative/nitrative tissue injury, independent of classical CB1/2 receptors.
Ethanol increases expression of macrophage metalloelastase (MMP‐12) via NADPH oxidase‐dependent ROS production in RAW264.7 macrophages
Authors: Kim Mi Jin , Nepal Saroj , Park Pil‐Hoon
Published: June 2021

The present study was undertaken to investigate the role of NADPH oxidase in ethanol‐induced MMP‐12 expression in RAW 264.7 macrophages. Here we found that ethanol induces significant increase in expression of NADPH oxidase subunits via p38 MAPK/NF‐κB pathway. Furthermore, knockdown of NOX2 (gp91phox), a membrane subunit of NADPH oxidase, by siRNA significantly inhibited ethanol‐induced ROS production, suggesting a critical role of NADPH oxidase in ROS production by ethanol. Ethanol treatment also causes significant increase in MMP‐12 expression at both mRNA and protein level. Inhibition of NOX2 by siRNA or by a selective pharmacological inhibitor prevented ethanol‐induced MMP‐12 expression. Taken together, these results demonstrated that NADPH oxidase‐derived ROS production plays a pivotal role in MMP‐12 expression in RAW 264.7 macrophages.
NOSH‐aspirin a dual nitric oxide‐and hydrogen sulfide‐releasing hybrid reciprocally regulates NF‐κB: S‐nitrosylation vs S‐sulfhydration
Authors: Kashfi Khosrow , Chattopadhyay Mitali , Kodela Ravinder , Song Teresa
Published: June 2021

Recently we reported the design of a hybrid aspirin with the ability to release nitric oxide (NO) and hydrogen sulfide (H2S), two important signalling molecules of physiological relevance, we have dubbed this new agent NOSH‐aspirin. It has been shown that NO can directly modify sulfhydryl residues of proteins through Snitrosylation and induce apoptosis. S‐sulfhydration of proteins by H2S may be analogous to that of protein S‐nitrosylation by NO leading to hydropersulfide (‐SSH) formation. In vitro: aspirin and NOSH‐ASA inhibited HT‐29 human colon cancer cell growth, the IC50s being >; 5000 μM and 0.05 ± 0.003 μM at 24h, respectively. NOSH‐aspirin reduced NF‐κB protein levels, and activated caspase‐3 enzyme in a dose‐and time‐dependent manner. Based on the biotin switch assay, which detects S‐NO and S‐SH proteins depending on presence or absence of ascorbate, we observed that a number of proteins from NOSH‐aspirin treated HT‐29 cells showed increased S‐nitrosylation and decreased S‐sulhydration. Pretreatment of the cells with carboxy‐PTIO and NaF, abrogated these effects. NOSH‐aspirin also S‐nitrosylated and S‐sulfhydrated NF‐κB p65 and denitrosylated and desulfhydrated caspase‐3 in a time‐dependent manner in these cells. In vivo: Liver lysates from rats treated with NOSH‐ASA showed dose‐dependent increases in S‐nitrosylation and decreases in S‐sulfhydration of NF‐κB p65. Thus, sulfhydration and nitrosylation both appear to regulate NF‐κB p65 in a reciprocal manner. These data provide a posttranslational mechanistic role for NO and H2S, and a rational for the chemopreventive effects of NOSH‐aspirin. Supported in part by NIH grant R24 DA018055