bluuebunny/crossref_metadata_2025 huggingface dataset:
Micronutrient Deficiencies Are Still Common among Chinese Children and Adolescents
Authors: Wang Huijun , Zhang Bing , Du Shufa , Popkin Barry
Published: June 2021

Protein‐energy malnutrition is rapidly decreasing in China, while few studies reported the status of micronutrient malnutrition. To explore the trends and associated factors of micronutrients intakes, including calcium, iron, zinc, Selenium, Vitamin A, vitamin B1, vitamin B2 and vitamin C, we analyzed dietary intake data among Chinese children and adolescents aged 4 to 17 years collected in an open cohort study, the China Health and Nutrition Survey (CHNS) from 1991 to 2011. We collected dietary data by using consecutive three 24‐hour recalls in combination with weighing methods. We assessed nutrient intake status by using the newly released estimated average requirement (EAR) for Chinese children. On average, daily calcium intake increased from 297.2 mg/day in 1991 to 310.2 mg/day in 2011 among boys and from 283.0 mg/day to 301.6 mg/day among girls during the same period. 11.4% to 18.5% of boys and 9.0% to 13% of girls reached the EAR levels for calcium. The average daily zinc intake of Chinese children decreased from 10.4 mg/day in 1991 to 9.6 mg/day in 2011, with only 21.5% of children reaching the EAR levels. Other micronutrient deficiencies are common. Multivariate analyses showed that micronutrient deficiencies are more common among girls than boys (p < 0.05) and among low income groups (p < 0.05). Micronutrients deficiencies should not be overlooked while most attentions are paid to childhood obesity in China.

This study was supported by the NIH (R01‐HD30880) and the Fogarty International Center, NIH (5D43TW007709).
Can Iodine Status be Predicted by Food Group Intake?
Authors: Lee Kyung Won , Song Won O. , Cho Mi Sook
Published: June 2021

Iodine deficiency leads to developmental failure, mental retardation, hypothyroidism, and goiter in children and adults. Dietaray assessement of iodine status is challenged as iodine contents in foods are not available. We aimed to assess if iodine status can be predicted by food group intakes in American adults. US adults (n=5,967, 蠅20y) in NHANES 2007‐12 with data on urinary iodine concentration (UIC), 24‐h dietary recalls, sociodemographic and lifestyle variables were included. All foods consumed are classified into 15 groups based on the USDA food code. Pearson correlations, χ2 and logistic regression were conducted to identify sociodemographic determinants and predictability of UIC by food group intake. A considerable percentage of women (38.8%), non‐Hispanic black (37.0%), 40‐59 year‐old (35.7%) and underweight (43.2%) adults were iodine deficient (UIC <100 ug/L). In contrast, the prevalence of excess iodine (UIC 蠅200 ug/L) is higher among men (38.8%), adults over 60 yr (38.3%), Mexican American (37.5%) and those who are obese (38.4%). UIC (ug/mg creatinine) correlated (+) with intake of milk, fish/seaweed, and breads groups, inversely (‐) with meat/poultry, eggs, other grain products, fruits, and beverages. The risk of iodine deficiency (<100 ug/L UIC) was lower (p<0.05) in consumers of milk, eggs, breads, fruits and vegetables. Subgroups with high iodine deficiencies need nutritional education to prevent iodine deficiencies and emphasize consumption of milk, eggs, breads, fruits, and vegetables, particulary in high risk sociodemographic subgroups.
Socially marketing micronutrients: Caregiver recall of a multi‐channel communications campaign in Bolivia
Authors: Harding Kimberly , Roche Marion , López de Romaña Daniel , Escobari Max , Merino Delsy
Published: June 2021

We designed and implemented a communications campaign in Bolivia, in partnership with the Ministry of Health and Sports, to increase demand and appropriate use of micronutrient supplements for children as part of the national Zero Malnutrition Program. The full campaign (TV and radio ads and print materials) was implemented in prioritized municipalities and only mass media was implemented in non‐prioritized municipalities (prioritization was based on population size, supplement coverage and anemia burden). We aimed to assess caregiver recall of the communications campaign, with and without community‐level print communication. 9 mo after campaign dissemination, we conducted a household‐level survey with caregivers of young children 6‐23 mo of age (n=363) in randomly selected prioritized (n=19) and non‐prioritized (n=18) municipalities. We administered a questionnaire on campaign exposure and related knowledge, attitudes and practices, and showed images of materials to facilitate recall. 30% of caregivers surveyed reported recall of the campaign, which was higher in urban than rural areas (37 vs 23%, p<.05) and higher in prioritized than non‐prioritized areas (37 vs 19%, p<.05). Ownership of TV was 84% and radio was 75%. Median daily use of each was 2 h. Overall recall of TV ads was 17% and radio was 9% (20 vs 13%, p=.1 and 7 vs 12%, p=.07 in prioritized and non‐prioritized areas, respectively). Recall of TV and radio ads was lower than expected, based on high TV and radio ownership and formative research findings which identified these mass media channels (stations and hours) as the preferred form of receiving health information.

Funding: Micronutrient Initiative
Status of the Fresh Fruit and Vegetable Program – 4 Years since Expansion
Authors: Ohri‐Vachaspati Punam , Turner Lindsey , Chaloupka Frank
Published: June 2021

The United States Department of Agriculture (USDA) provides funding for the Fresh Fruit and Vegetable Program (FFVP) across the US with the goal to create healthier food environments in lower‐income elementary schools. Four years of data collected from 2009‐10 to 2012‐13 by the Bridging the Gap program from more than 2,200 public elementary schools participating in the National School Lunch Program were used to examine trends in FFVP participation. In 2012‐13, 36% of schools participated in the program, compared to 26% in 2009‐10. Schools with more low‐income students, with smaller enrollment, having a dietitian on staff and participating in the USDA's Team Nutrition program had significantly greater odds of participation in the FFVP. Presence of a full service kitchen was not associated with FFVP participation. Schools with dietitian on staff saw a 41% increase in participation from 2009‐10 to 2012‐13, compared to a 32% increase in schools with no dietitian. Although Team Nutrition schools started off with lower rates of participation, participation in non‐Team Nutrition schools increased at a higher rate in the last two years of reporting. Rates of participation have increased at similar rates among schools of different sizes. FFVP participation has increased in elementary schools over four years and additional resources such as having a dietitians on staff, participation in USDA's Team Nutrition programs, and a smaller school size are likely facilitators for FFVP participation.
Effects of a World Food Program‐Sponsored Malnutrition Intervention Program on Undernutrition and Stunting in La Estancia, El Salvador
Authors: Rosales Rachael , Gosselin Kerri , Kasper Jennifer
Published: June 2021

Background

6% and 19% of Salvadoran children 0‐59 months of age are moderately or severely underweight and stunted respectively. Estancia consists of nine rural villages Northeastern El Salvador; 33% of population lives in extreme poverty; housing, safe water and sanitation are lacking; food insecurity is among highest nationwide.

Methods

Convenience sample of 190 children under two years age received education talks, anthropometric measurements, and distribution of WFP Super Cereal plus, beans, rice, and vegetable oil. Data entered into WHO Anthro database. Children were classified as normal (>‐2SD) or malnourished (= or < ‐2 SD). Analysis conducted using STATA.

Results

At enrollment, mean age was 11.2 months, 45% female, 29 (15.3%) had WAZ <=‐2, 69 (36.3%) had HAZ <=‐2. Average length of intervention was 179 days. After supplementation, 22 (11%) had WAZ <=‐2 and 86 (45%) had HAZ <=‐2.
Limitations: Small sample size, lack of attribution, possible measurement and data entry errors, lack of health data. Likely distribution of nutritional supplements among all family members, and Super Cereal's incomplete coverage of children's RDA, potentially dilute effect of nutritional supplement on improving nutritional status.

Conclusions

Baseline acute undernutrition and stunting rates nearly double country‐wide prevalence, highlighting Estancia's higher burden of disease. Older children were more likely to be stunted. WPF nutritional supplement decreased acute undernutrition, but was insufficient to reduce chronic malnutrition and stunting.
Children in Child Care Centers that Participated in the Healthy Apple Award (HAA) Program in San Francisco had Greater Decreases in BMI Percentile than Children in Other Local Child Care Centers
Authors: Stookey Jodi , Evans Jane , Nelson Amy , Chan Curtis
Published: June 2021

First Lady Michelle Obama's Let's Move! Child Care initiative aims to improve nutrition and physical activity (PA) practices in child care sites to promote healthy childhood weight, nationwide. This analysis tested if a new program in San Francisco, the HAA program, designed to support implementation of Let's Move!, impacts childhood weight, locally. Child care centers served by the Child Care Health Program (CCHP) were randomly selected for early (2013) or lagged (2014) invitation to the HAA. Child care centers that enrolled in 2013 (CCHP+HAA) were offered a self‐assessment modeled on Let's Move! and the University of North Carolina's Nutrition and Physical Activity Self Assessment in Child Care (NAP SACC) in Spring, technical assistance for goal setting and staff trainings about drinking water and PA in Summer, and re‐assessment and award recognition in the Fall. All CCHP+HAA (n=18) and CCHP‐only (n=20) centers were offered child‐sized water pitchers in Fall 2013, health worker assessment of drinking water and PA resources and BMI screening for all enrolled children in Fall 2013 and Spring 2014 (n=871, 3‐4y). In CCHP+HAA centers, the proportion of children with access to visible and self‐serve drinking water (61% to 83%) and a PA curriculum (36% to 50%) increased. Corresponding changes in CCHP‐only centers were 63% to 73% and 45% to 34%. Mean BMI percentile decreased by 2.4 (1.2) more percentage points in CCHP+HAA vs. CCHP‐only centers (p<0.05). The results suggest that the HAA program supports local implementation of Let's Move!.
Nutritional Knowledge and Dietary Behaviors and Intakes of Vietnamese Marriage Immigrant Women According to Receiving of Nutritional Education
Authors: Hwang Ji‐Yun , Joe Mee‐Young
Published: June 2021

This study was to investigate nutritional knowledge and dietary behaviors and intakes of Vietnamese marriage immigrant women according to receiving of nutritional education. A total of 233 Vietnamese marriage immigrants were recruited from public health centers, and multicultural family support centers in Seoul Metropolitan areas, and the Seoul Immigration Office. The methods included anthropometric measurements and face‐to‐face interview by a structured questionnaire. The mean weight and BMI of those who had experienced nutritional education were higher than those without such experience. With regard to dietary management, those with nutritional education showed a more balanced diet and higher total scores on the Mini Dietary Assessment compared to those without nutritional education (p<0.001). For nutritional knowledge, subjects with nutritional education showed higher scores on questions about function of cereals (p=0.010) and meat and fish (p=0.010), as well as a higher overall score (p=0.004) than those without. With regard to food intake, there were significant differences in nutritional intakes of iron (p=0.009), sodium (p=0.008), vitamin A (p=0.029), vitamin B2 (p=0.047), vitamin C (p=0.012), and cholesterol (p=0.037). The subjects having experience with nutritional education had more balanced dietary habits, and nutritional knowledge, and greater food and nutrient intake compared to the subjects without such experience. Systematic and continuous nutritional education should be provided to establish a healthy diet for marriage immigrant women and her multicultural family members. This study was supported by the grant from National Research Foundation of Korea (2011‐0014986)
The Unfolding Tail of CHIP Mutation in Gordon Holmes Syndrome
Authors: Rubel Carrie , Soss Sarah , McDonough Holly , Chazin Walter , Patterson Cam , Schisler Jonathan
Published: June 2021

Gordon Holmes Syndrome (GHS) is a rare neurodegenerative disorder characterized by cerebellar ataxia with hypogonadism. The pathophysiological mechanisms and genetic causes of GHS are largely unknown and therapies limited. We identified a homozygous mutation (c.737C→T, p.Thr246Met) in STIP1 homology and U‐box containing protein 1 (STUB1), the gene that encodes for C‐terminus of Hsp70 Interacting Protein (CHIP) in siblings that presented with GHS. CHIP plays a central role in regulating protein quality control as both an E3 ligase and molecular chaperone. Human mutations of CHIP are increasingly linked to ataxias, but the molecular mechanisms underlying this disease pathology remain undefined. Biophysical and cell culture studies reveal T246M CHIP has a disorganized Ubox domain and exists mostly as aggregates. Consequently, T246M CHIP exhibits loss of E3 ligase activity, but unexpectedly, the amount of T246M CHIP interacting with substrates, E2 enzymes and molecular chaperones is higher than wild‐type CHIP. We hypothesize that the sequestering of CHIP interacting proteins drives disease pathology. Interestingly, T246M CHIP maintains some chaperone activity as measured by its in vitro potentiation of AMPK and translocation/activation of HSF1 in cells. Thus the T246M mutation disrupts CHIP structure and function resulting in a dominant negative protein impairing ubiquitin‐dependent protein degradation. We developed a T246M CHIP mouse model of GHS and present preliminary data consistent with our hypothesis that T246M CHIP functions as a dominant negative protein causing GHS.
Elucidation of Resolvin and Protectin Sulfido‐Conjugated Mediators: New Pro‐Resolving and Tissue Regenerative Pathways
Authors: Ramon Sesquile , Dalli Jesmond , Norris Paul , Colas Romain , Serhan Charles
Published: June 2021

Resolution of inflammation is an active cellular and biochemical process mediated by autacoids. Here, we investigated self‐limited infectious exudates for novel bioactive chemical signals in tissue regeneration and resolution. Using spleens from E. coli infected mice, self‐resolving infectious exudates, human spleens and blood from sepsis patients we identified two new families of potent molecules. Characterization of their physical properties and isotope tracking demonstrated that the bioactive structures contained a docosahexaenoate backbone and sulfido‐conjugated triene or tetraene double bond systems. Activated human phagocytes converted 17‐hydro(peroxy)‐4Z,7Z,10Z,13Z,15E,19Z‐docosahexaenoic acid to these bioactive molecules. Regeneration of injured planaria was accelerated with nanomolar amounts of 16‐glutathionyl, 17‐hydroxy‐docosahexaenoic acid and 16‐cysteinylglycinyl, 17‐hydroxy‐docosahexaenoic acid (coined protectin conjugates in tissue regeneration) as well as 8‐glutathionyl, 7,17‐dihydroxy‐docosahexaenoic acid and 8‐cysteinylglycinyl, 7,17‐dihydroxy‐docosahexaenoic acid (coined resolvin conjugates in tissue regeneration). Each protectin and resolvin sulfido‐conjugate stimulated human macrophage bacterial phagocytosis, phagolysosomal acidification and efferocytosis. Together these results identify two novel pathways and structural elucidation of new resolution moduli. These pathways and mediators display host protective, pro‐resolving and tissue regenerative actions, properties essential for tissue homeostasis.
Ion Efflux and Influenza Infection Trigger NLRP3 Inflammasome Signaling in Human Dendritic Cells
Authors: Fernandez Melissa , Miller Elizabeth , Krammer Florian , Greenbaum Benjamin , Bhardwaj Nina
Published: June 2021

The NLRP3 inflammasome, a multi‐protein complex, is an essential intracellular mediator of anti‐viral immunity. In murine dendritic cells (DCs) this complex responds to ion efflux and influenza A virus (IAV) infection to activate caspase‐1 mediated proteolysis of interleukin (IL)‐1β and IL‐18 into biologically active cytokines. However, the presence and function of the NLRP3 inflammasome in human DCs in response to various triggers, including viral infection, has not been clearly defined. Here, we delineate the contribution of the NLRP3 inflammasome to the secretion of IL‐1β, IL‐18, and IL‐1α by human DCs (monocyte‐derived DCs (MDDCs) and primary conventional (cDC)). Activation of the NLRP3 inflammasome in human DCs by various synthetic activators resulted in the secretion of IL‐1β, IL‐18, and IL‐1α, and induction of pyroptotic cell death. Cellular IL‐1β release depended on potassium efflux and the activity of proteins NLRP3 and caspase‐1. Similarly, IAV infection of DCs resulted in priming and activation of the NLRP3 inflammasome and secretion of IL‐1β and IL‐18 in an M2 and NRLP3 dependent manner. The magnitude of priming by IAV varied amongst different strains and inversely corresponded to type I IFN production. To our knowledge this is the first report describing the existence and function of the NLRP3 inflammasome in human DCs and the ability of IAV to prime and activate this pathway in human DCs, with important implications for viral pathogenesis.
Histone Variant MacroH2A1 Marks Liver Aging and Orchestrates the Escape from Senescence Induced by DNA Hypomethylation in Hepatocellular Carcinoma
Authors: Vinciguerra Manlio , Borghesan Michela , Rappa Francesca , Panebianco Concita , Cappello Francesco , Pata Illar , Buschbeck Marcus , Sedivy John , Fusilli Caterina , Mazza Tommaso , Pazienza Valerio
Published: June 2021

The epigenetic basis of age‐associated progression of liver diseases towards hepatocellular carcinoma (HCC) is unclear. MacroH2A1 is a variant of histone H2A1, present in the two isoforms, with fundamental roles in cell homeostasis. MacroH2A1 is a marker of senescence associated heterochromatic foci (SAHF) and synergizes with DNA demethylating chemotherapic agent 5‐aza‐2'‐deoxycytidine (5‐aza‐dC) in silencing tumor suppressor genes in human fibroblasts.

We show that protein levels of macroH2A1 isoforms are increased in the livers of old rodents and humans, and in human HCC tissue. Human HCC cells overexpressing macroH2A1 escape a 5‐aza‐dC‐induced senescent phenotype, as determined by cell proliferation/migration/cell cycle assays and b‐gal staining, and exhibit a synergistic global DNA hypomethylation. Deletion of macroH2A1 had opposite, pro‐senescence effects in presence of drug dependent DNA hypomethylation. System analysis of ChIP‐Seq combined with RNA‐Seq data revealed complex macroH2A1 genome occupancy correlated with anti‐senescence transcriptional patterns.

We propose macroH2A1 as a strong candidate target for epigenetic‐ and senescence‐based HCC chemotherapy.
Umbilical Cord Mesenchymal Stromal Cell Isolation Expansion and Validation for GMP Compliance
Authors: Smith J Robert , Pfeiffer Kyle , Delzeit Jennifer , Akel Salem , McGuirk Joseph , Weiss Mark
Published: June 2021

Developing methods for isolating, expanding and validating umbilical cord mesenchymal stromal cells (UC‐MSCs) with Good Manufacturing Practice (GMP) compliance will allow quicker clinical translation from in‐vitro and preclinical findings. MSCs are used in clinical trials for their abilities to reduce inflammation by modulating the immune system and differentiate into several tissues, such as adipose, cartilage and bone. UC‐MSCs provide a limitless supply and higher quality cells than MSCs from other tissues, however clinical translation requires GMP compliant SOPs, which have not yet been developed. We developed a closed system isolation method for UC‐MSCs using Miltenyi dissociator tubes coupled with enzymatic digestion, Red Blood Cell lysate treatment, and filtering to isolate MSCs. Expansion methods analyzed a non‐xenogeneic medium supplemented with human platelet lysate (HPL). The MSCs were validated by analysis with flow cytometry, multilineage differentiation and Colony Forming Unit‐Fibroblast assay (CFU‐F). The new method provides a higher cell yield than previous methods and eliminates cumbersome steps including mincing and blood vessel removal. MSCs had the highest growth rate in media supplemented with the highest concentration of HPL. Flow Cytometry data shows > 95% positive for MSC markers and CFU‐F data shows high frequency of CFU's. This method provides a platform that can be transferred easily to a GMP facility to speed the clinical translation of UC‐MSCs.
Blended Learning in The Context of Over‐The‐Counter Drugs: An Approach to Medical Pharmacology Education
Authors: McHugh Douglas , Macica Carolyn
Published: June 2021

Medical students often report anxiety and difficulty with assimilating the extensive pharmacology content required of them. To mitigate this we created blended learning events for students to build cognitive schemas into which they will integrate therapeutic drug knowledge. We developed an online series of modules that introduced pharmacokinetics and pharmacodynamics into the first year curriculum. Students were assigned to smallgroups of ~5 individuals. Modules included a case vignette, online didactic teaching video, over‐the‐counter (OTC) drug wiki assignment and cumulative concept map. Wiki assignments encouraged active learning with students answering 1 of 5 questions and providing at least 1 comment in response to other student posts. The questions were designed to not have neat, easily 'Google‐able' answers and were delivered in the context of common OTC medications that students were likely to have experienced themselves. Wiki assignments also provided a safe environment for intellectual risk‐taking and promotion of adaptive learning behaviors. In so doing, they fostered students' ability to reason scientifically based on their understanding of pharmacokinetic and pharmacodynamics principles, and provided a space for making mistakes and tactful peer correction/teaching. The modules integrated with lectures delivered throughout the systems biology curriculum of year 1. Together, these blended learning components provided a format for the students to develop a conceptual framework that fostered mechanism‐based learning rather than rote memorization. Preliminary formative and summative assessment data suggest an improvement in pharmacology performance.
A Mentorship Toolkit for Teachers and Researchers in Pharmacology and Other Biomedical Sciences in Developing Countries
Authors: Kwanashie Helen
Published: June 2021

Formal mentorship programs are frequently not available in higher education institutions in the less developed countries. However, the demonstrated benefits of mentoring to mentees, mentors and institutions, provide strong rationale for its more widespread application. Younger pharmacologists and other early‐career biomedical scientists (especially women), are identified primary target beneficiaries. This paper describes simple logistics (objectives, formats, guidelines for monitoring and evaluation, mentoring agreement and progress record forms, etc) that could facilitate the setting‐up of mentorship programs in resource‐poor settings. Special attention is paid to cross‐gender mentoring, including the advantages and disadvantages of women mentoring women. Application of internet resources (such as email and social media) as well as cross‐cultural mentorships are also incorporated. The output is 'A Mentorship Toolkit for Teachers and Researchers in the Biomedical Sciences in Developing Countries'; and is ready be field‐tested in practice. Although the overall goal of the current presentation is to improve Pharmacology Education across West Africa, the mentorship toolkit can readily be adapted to suit other disciplines, and be used even in the more developed world.
Global Curve Fitting Approach to Receptor Allosterism
Authors: Zhang Rumin , Kavana Michael
Published: June 2021

G‐protein coupled receptors (GPCRs) are known to be modulated by both orthosteric agonists and positive/negative allosteric modulators (PAMs/NAMs). Proper experimental design and data analysis for dual dose dependence between agonist and PAM or NAM are critical to elucidate key parameters for understanding molecular mechanism and structure‐activity relationship (SAR) in drug discovery. Toward that end, we have developed a global curve fitting approach to analyze simultaneously all curves in dual dose dependence data of receptor allosterism to report all key parameters. A general equation based on operational model of receptor allosterism is reformulated in GraphPad Prism to allow global curve fitting. Specific stepwise instructions with graphic illustrations will be outlined.
PDI via Nrf2 Contributes to the Regulation of Renal AT1 Receptor Function and Blood Pressure
Authors: wang xitao , Asghar Mohammad
Published: June 2021

We recently reported that age‐associated oxidative stress plays a causal role in high renal angiotensin AT1 receptor (AT1R) function and blood pressure (BP) in the aging Fischer Brown Norway (FNB) rats. This indicates that redox balance is crucial for normal AT1R function and BP. Our objective was to understand the mechanism responsible for the higher oxidative stress in the aging kidneys contributing to impaired AT1R function and high BP in the aged FBNs. Keap1‐Nrf2 system is the master cytoprotective regulatory pathway in the kidney against oxidative stress. Nrf2 transcription factor protects kidney from oxidative stress by activating the transcription of diverse antioxidant and detoxifying enzymes. Kelch‐like ECH‐associated protein 1 (keap1) constitutively represses Nrf2 activity by sequestering Nrf2 in cytosol. We hypothesize that protein disulfide isomerase (PDI), an enzyme involved in isomerization reaction, regulates keap1 and Nrf2. Nrf2, keap1 and PDI were determined by Western blotting and/or immunohistochemistry in both the kidney cortical tissue of FBNs and human kidney 2 (HK2) cells. We found that the aging kidney had low levels of PDI, which was associated with low nuclear levels of Nrf2 and high cytosolic levels of keap1 in the aging FBNs. Moreover, PDI inhibition by bacitracin in HK2 cells significantly reduced Nrf2 accumulation in the nuclei. PDI siRNA treatment in HK2 cells decreased the level of PDI while increased the level of keap1. These data suggest that PDI via keap1 and Nrf2 may be involved in redox balance, contributing to the regulation of AT1R function and BP in aging.

Source of Research Support: NIH/NIA, AG039856
CB1 receptor transgenic mice in the cannabinoid triad: a novel approach to assess in vivo efficacy of CB1 ligands.
Authors: Grim Travis , Negus Steve , Lichtman Aron
Published: June 2021

The abuse of structurally diverse synthetic cannabinoids has resulted in untoward effects in users and concomitant spike in emergency room visits, which signify a significant public health concern. Synthetic cannabinoids bind and activate cannabinoid CB1 receptors, which are responsible for the psychomimetic effects of the primary active constituent of marijuana, D9‐tetrahydrocannabinol (THC). While THC is classified as a partial agonist, many of the synthetic cannabinoids have been identified as full agonists. While various in vitro methods assess CB1 receptor efficacy (e.g. [35S]GTPgS binding, cAMP production), only drug discrimination procedures are currently utilized to determine in vivo efficacy. Here, we assess efficacy utilizing a cumulative dose‐response procedure of several cannabinoids (WIN55,212‐2, CP55,940, CP47,497, and THC) on CB1 receptor‐mediated pharmacological effects (i.e., catalepsy, hypothermia, and thermal antinociception) in CB1 (+/+), (+/‐), and (‐/‐) mice. As expected, these compounds produced little or no effects in CB1 (‐/‐) mice, consistent with the premise that these effects are CB1 receptor mediated. Each of these compounds produced maximum effects in CB1 (+/+) mice, but (+/‐) mice displayed differential sensitivity to these CB1 receptor agonists that was positively associated with their in vitro efficacy. Analysis of potency ratios between CB1 (+/+) and (+/‐) mice revealed a rank order of efficacy (from highest to lowest) of WIN55,212‐2 > CP55,940 > CP47,497 > THC. Together, these experiments demonstrate a quick and accurate method to determine in vivo efficacy, and this design correlates well with prior in vitro efficacy determinations in [35S]GTPgS binding assays.
Hypothermic Effects of Δ9‐THC and Nicotine in Kynurenine‐3‐Monooxygenase (KMO) Knockout Mice
Authors: Cocke Morgan , Moerke Megan , O'Connor Jason , McMahon Lance
Published: June 2021

Kynurenic acid, a metabolic product of the tryptophan‐kynurenine synthesis pathway, is both a competitive antagonist at the glycine co‐agonist site of NMDA receptors and a negative allosteric modulator at α7 nicotinic acetylcholine receptors (nAChRs). α7 nAChRs are important for mediating the effects of nicotine, but have also recently been implicated in the effects of Δ9‐tetrahydrocannabinol (Δ9‐THC), the primary psychoactive component of Cannabis. Transgenic mice lacking kynurenine‐3‐monooxygenase (KMO) exhibit increased levels of kynurenic acid and thus presumably decreased function of α7 nAChRs. The current study examined the extent to which the hypothermic effects of nicotine and Δ9‐THC are modified in KMO knockout mice (n=7) as compared to wild‐type C57BL/6J controls (n=7). Δ9‐THC (10 mg/kg) produced a maximum decrease in temperature of 3.0°C at 60 min; hypothermia was no longer evident at 240 min. Nicotine (1.78 mg/kg base) produced a maximum hypothermic effect of 7.2°C at 30 min; this effect was no longer evident at 90 min. The magnitude of effect and the time course of hypothermia produced by Δ9‐THC and nicotine were not significantly different between KMO knockout and wild‐type mice. These data suggest that at least some of the in vivo effects of Δ9‐THC and nicotine are not affected by differences in endogenous levels of kynurenic acid. Supported by USPHS grant DA25267 and MH090127.
Neutralization of Aβ‐amyloid induced toxicity by oligomer 'trapping'
Authors: Multhaup Gerhard , Barucker Christian , Althoff Veit , McKinney Anne
Published: June 2021

Objectives

The transmembrane sequence (TMS) of APP (Aβ residues 29 to 49) is a key determinant of Aβ42 oligomerization and toxicity, with G33 being the critical residue (Harmeier et al., 2009). A systematic analysis of Aβ‐lipid interactions revealed that residues 29 to 49of the neurotoxic Aβ42 bind more strongly to sphingomyelin (SM), GM1, phosphatidylcholine (PC), and a mixture of neuronal lipid membranes (BTLE) than its non‐toxic counterparts Aβ40 and Aβ42 G33I. As the contact site likely represents a drug target site, potential molecules interfering with Aβ‐lipid interactions should enable the design of amyloid selective therapeutics.

Methods

Surface plasmon resonance (SPR), Size‐exclusion chromatography (SEC), organotypic slice cultures, MALDI‐MS, electron microscopy, and Drosophila melanogaster as an in vivo model for AD.

Results

To date, we have found that an eight amino acid peptide (a so‐called “Aβ42‐oligomer interacting peptide” ‐ AIP) with alternating hydrophobic and hydrophilic amino acids neutralizes Aβ42‐induced neurotoxicity by binding to preferentially highly toxic Aβ tetra‐/hexamers. Our data indicates (i) that co‐administration of AIP with Aβ42 prevents the loss of synaptic spine density and rescues long‐term potentiation (LTP) in organotypic slice cultures, and (ii) that the AIP exhibits protective effects in our in vivo model for AD.

Conclusions

Membrane regions with a high percentage of SM, such as “lipid rafts” could be of particular importance for the interaction with Aβ42 and may play a crucial role in mediating amyloid in vivo toxicity. The development of compounds that interfere with toxic Aβ‐lipid interactions may have a preventive and therapeutic potential.
Reducing NADPH Oxidative Stress following Acute Neurotrauma
Authors: Lucke‐Wold Brandon , Robson Matthew , Logsdon Aric , Turner Ryan , Rosen Charles
Published: June 2021

1.7 million traumatic brain injuries occur each year in the USA. Utilizing a validated rodent blast model, we investigated how NADPH oxidase expression and associated oxidative stress contributes to cellular apoptosis following blast injuries. We found a visible increase in NADPH Nox4 following single blast injury in Sprague Dawley rats. Interestingly, Nox4 was also increased in post‐mortem human samples obtained from deceased athletes. Nox4 activity led to an increase in superoxide formation. Lipoic acid decreased Nox4 when given i.p. 10mg/kg 5‐minutes after blast exposure. Lipoic acid also significantly decreased the subsequent formation of superoxide. Lipoic acid increased the anti‐apoptotic markers Bcl‐2 (t = 3.079, p<0.05) and heme oxygenase 1 (t = 8.169, p<0.001) compared to blast injured animals. Two weeks following the last blast exposure (6 blasts total over 2 weeks), the lipoic acid treatment group had significantly reduced pro‐apoptotic markers Bax (t=4.483, p<0.05), caspase 12 (t=6.157, p<0.001), and caspase 3 (t=4.573, p<0.01) compared to blast injured animals. Lipoic acid treated animals also had significantly reduced tau hyperphosphorylation. Tissue from the NFL player had increased Bax expression (t=5.424, p<0.01) compared to control. A significant difference in impulsive‐like behavior between experimental animal groups was seen 7 days after repetitive blast injury (F(2,21)=4.327, p<0.05). Rats exposed to repetitive blast displayed increased time in the open arm compared to control (t=3.632, p<0.05), but lipoic acid ameliorated this impulsive‐like behavior (t=3.573, p<0.05). Lipoic acid warrants further investigation as a therapeutic for the treatment of acute neurotrauma.
Noribogaine is a Mixed Agonist/Antagonist Opioid Ligand with Profound Functional Selectivity
Authors: Maillet Emeline , Milon Nicolas , Fishback James , Schürer Stephan , Heghinian Mari , Garamszegi Nandor , Mash Deborah
Published: June 2021

Noribogaine is the primary metabolite of the anti‐addictive substance ibogaine, which modulates opiate analgesic activity and the components of drug addiction in animal models at brain concentrations of 0.5‐15 µM. In this study, molecular activities of noribogaine at mu (OPRM) and kappa (OPRK) opioid receptors were characterized. Noribogaine was a moderately potent antagonist of the OPRM G‐protein and β‐arrestin signaling pathways (20 µM; 48 µM). Noribogaine was a partial agonist at the OPRK G‐protein pathway, activating at 75% the maximal efficacy of Dynorphin A (Dyn‐A) at a potency of 9 µM, and had weak inhibitory properties (40 µM, 25% against Dyn‐A). Noribogaine was a biased agonist and poorly activated the OPRK β‐arrestin pathway at 12% of Dyn‐A maximal efficacy. In turn, noribogaine was able to functionally inhibit Dyn‐A‐induced β‐arrestin recruitment (Dyn‐A EC50: 82 nM) at physiologically relevant concentrations (IC50 of 1.45 µM at 370 nM Dyn‐A). Computational simulations indicated that noribogaine may bind to the orthosteric morphinan binding site of the receptor. This study clarifies the action of noribogaine at modulating opioid receptor function, uncovering explanatory mechanisms as well as new avenues of therapeutic development.
Pharmacogenomics of Methotrexate in Rheumatoid Arthritis Patients
Authors: Hagras Magda , Al Attar Suzan , Abuzenadah Adel , Fathaldin Omer , Aseri Ahmed , Sulaiman Mansour , Raddadi Rajaa , Goeb Vincent
Published: June 2021

Patients with rheumatoid arthritis receiving methotrexate show different responses to treatment. Therefore, this study investigate whether changes in efficacy and toxicity were correlated with allele frequency of different enzymes dealing with methotrexate. Sixty Adult Saudi patients of both sex diagnosed with rheumatoid arthritis receiving methotrexate were recruited, from King Abdulaziz University Hospital, starting from December 2013 to Nov. 2014. Blood sample was withdrawn for genetic analysis of rs717620, rs1045642, rs1232027 and rs1650697 using the Taqman RT‐PCR. Effectiveness and toxicity were evaluated.
Forty nine were females, 40.9 ± 9.1 year with disease duration 4.7±5.5 yr, duration of MTX was 36.1 ± 29.6 months and the MTX dose was 12.7±3.7 mg/wk. The mean DAS at baseline was 3.19±1.6.
Results revealed that Allele frequency for Homozygous G of ABCC2 (rs717620) is 60%, hetero CTfor ABCB1 (rs1045642) is 56.7%, Allele frequency for Hetero GA of DHFR (G>A) (rs1232027) is 51.7%, Homo C for DHFR (C>T) (rs1650697) is 65.0%. The association of allele with nausea, vomiting, hair loss, photosensitivity, chest symptoms, menstrual symptoms, bone ache and deformities were not significant. In conclusion: No correlation between toxicity and allele frequency.
Characterization of a Novel “Thaw and Go” Cell‐based Transporter Model for Studying SLC Transporters
Authors: Wang Jie , Bourgea Joanne , Cooper Kirsten , Patten Christopher , Li Na
Published: June 2021

Objective

Development and characterization of a novel cell‐based transporter model to study regulatory agencies (USFDA/EMA) recommended and industry important SLC transporters.

Methods

Cryopreserved HEK293 cells which transiently overexpress OATP1B1*1a (wild‐type), OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1, MATE2‐K, OATP1A2, OATP2B1, PEPT1, PEPT2, NTCP and empty vector, respectively, were thawed and plated on 24‐well Poly‐D‐Lysine coated plates, and then refed with plating media in the presence or absence of 2mM sodium butyrate at 3‐4 hours post plating. Uptake assays were performed at 24 hrs post plating. Following two washes, cells were pre‐equilibrated with HBSS buffer for 10min, MATE1 and MATE2‐K cells require extra incubation with HBSS buffer containing NH4Cl. The cells were then incubated with substrate or inhibitor for desired amount of time. After the assay was terminated, the cells were lysed and subjected to analysis.

Results

The novel SLC transporter model, including OATP1B1*1a (wild‐type), OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1, MATE2‐K, OATP1A2, OATP2B1, PEPT1, PEPT2, and NTCP cells, was characterized for time and substrate concentration‐dependent uptake (Km and Vmax determinations) using prototypical substrates as well as known drug substrates. The model was also validated using selected prototypical transporter inhibitors recommended by regulatory agencies. The kinetic profiles of the selected substrates and IC50 of selected inhibitors were comparable to the published literature and also consistent across independent laboratories. As an example, OATP1B1*1a Km comparison using various substrates is demonstrated below.

OATP1B1*1a Substrates

Estradiol‐17β‐Glucuronide
Estrone‐3‐Sulfate
Rosuvastatin
Fluorescein Methotrexate

Km (µM) using Corning® OATP1B1*1a Cells
6.2
0.4
14.2
5.2

Km (µM) reported in literature
6.3
0.46
13.1
3.8

Conclusions

The data demonstrated this new SLC transporter model is a validated and compliant in vitro tool to study drug interactions with SLC transporters recommended by regulatory agencies (USFDA/EMA) and industry. This novel cell‐based transporter model eliminates the need for intensive cell culture and maintenance, as well as providing consistent lot‐to‐lot performance.
Effects of a Fusarium Mycotoxin, Beauvericin, on Bovine Granulosa Cell Proliferation and Steroid Production
Authors: Albonico Marco , Schutz Luis , Caloni Francesca , Cortinovis Cristina , Spicer Leon
Published: June 2021

Beauvericin (BEA) is a common wheat and maize contaminant produced by Fusarium species. Potential reproductive effects of BEA have not been investigated previously. Thus, objectives of this study were to determine if BEA can affect granulosa cell proliferation and steroid production. Ovaries of cattle were collected at a local slaughterhouse and follicular fluid was aspirated from small follicles (1‐5 mm) to isolate granulosa cells. Granulosa cells were cultured for 2 days in 10% fetal bovine serum followed by 2 days in serum‐free medium containing 500 ng/ml of testosterone (as an estradiol precursor), 30 ng/ml of follicle stimulating hormone, and insulin‐like growth factor 1 (IGF1; 0 or 30 ng/ml) with BEA (0, 3, or 30 μM). This experiment was replicated with three independent pools of cells and treatments applied in triplicate for each pool. Data were analysed as a 2 x 3 factorial ANOVA. Both IGF1 and dose of BEA significantly affected variables measured. IGF1 increased (P < 0.05) cell numbers and estradiol and progesterone production. BEA at 3 and 30 μM significantly inhibited basal and IGF1‐induced granulosa cell numbers by 80%. BEA did not affect basal estradiol or progesterone production at either dose, but BEA at 3 μM significantly inhibited IGF1‐induced estradiol and progesterone production by 77% and 71%, respectively. In conclusion, the present study demonstrates that significant toxicological effects of BEA are observed in a bovine granulosa cell model, and suggests that Fusarium mycotoxins such as BEA may have the potential to impair reproductive function in cattle.
Lactate Dehydrogenase is Unrelated to Lactic Acid Production in MDA‐MB‐231 Human Breast Cancer Cells
Authors: Mack Nzinga , Mazzio Elizabeth , Bauer David , Soliman Karam
Published: June 2021

Background

A well‐known characteristic of tumor cells is the propensity to produce lactic acid under normoxia and hyperoxia, also referred to as the Warburg effect. High levels of lactic acid are believed to be associated with elevated expression of lactate dehydrogenase A (LDHA), a gene upregulated in diverse aggressive malignancies.

Methods

In this study, ShRNAs were used to permanently knock down the expression of LDHA in human breast cancer MDA‐MB 231 cells. LDHA knockdown was confirmed by Western Blot analysis. Dual analysis of lactic acid production was done via colormetric assay and HPLC, where the data shows no reduction in lactic acid production in knockdowns versus controls. Protein expression of various subtypes of LDH was investigated in order to determine if other subtypes were upregulated.

Results

There was no change in the protein expression of other subtypes of LDH.

Conclusion

Lactate production in LDHA knockdowns is unrelated to the LDH protein, which calls into question the relationship between glycolysis and the upregulation of lactic acid production in MDA‐MB231 cells. Further analysis will require genomic or proteomic profiling to discover shifts in genes or proteins that are responsible for this sustained production of lactate.
Modeling TRPV1, a Detector of Thermal and Chemical Stimuli, Producing Pain: No Capsaicin Sensation
Authors: Strandberg S , Fleischmann S , Barrett E , David J , Feller C , Gottfried S , Hairston A , Schauer E , Siy T , Soczka S , Thew M , Zagloul M , Ahmad A , Ahmad Y , Carrig M , Dzwierzynski J , Rooney C , Ryan R , Scherrer C , Zappia Katherine , Weyer Andy
Published: June 2021

According to the Institute of Medicine, 100 million Americans suffer from chronic pain every year and the US spends over $500 billion trying to treat them. Pain begins as a stimulus that is detected by nociceptors, which are nerve fibers responsible for the detection of noxious mechanical, thermal, or chemical stimuli that give rise to pain sensations. These nociceptors transmit pain signals from the periphery to neurons in the spinal cord and brain. The Transient Receptor Potential Vanilloid 1 (TRPV1) is a nociceptive ion channel activated by capsaicin (the spicy component of hot peppers), heat, and endogenous pain molecules. Therefore, creating an inhibitor that partially blocks TRPV1 could treat chronic pain. The amino acids in the active site of TRPV1 are Y511, S512, M547, and T550. In addition, E600 controls the selectivity filter at the top gate of the channel and the hydrophobic seal mediated through I679 controls the lower gate. When capsaicin binds to the channel, a conformational change occurs that pulls the I679s on each subunit away from each other, opening up the lower gate. Understanding how activation of TRPV1 occurs may lead to the discovery of novel inhibitors of TRPV1 to help treat those suffering from chronic pain and reduce healthcare spending. The Divine Savior Holy Angels SMART (Students Modeling A Research Topic) Team modeled TRPV1 in a partially activated state using 3D printing technology. Program supported by a grant from NIH‐CTSA.
Single Cell Analysis of Embryonic Stem Cell‐Derived Vascular Endothelial Sub‐phenotypes
Authors: Madfis Nicole , McCloskey Kara
Published: June 2021

A well‐formed and robust vasculature is critical to the health of most organ systems in the body. To that end, our laboratory has directed the differentiation of vascular subphenotypes from stem cells for a number of applications. Our step‐wise methodology outlining vascular endothelial cell (EC) differentiation from embryonic stem cells (ESC) in sera‐free conditions demonstrates a range of endothelial heterogeneity and vascular functionality in these cultures. Vascular endothelial subphenotypes analogous those EC in angiogenic (tip and stalk EC) and quiescent blood vessels (phalanx EC) were in our 2D in vitro cultures. Here, distinct “tip/stalk‐like” and “phalanx‐like” EC subphenotypes were analyzed for cell surface marker expression using FACS analysis and immunohistochemistry. We used a nine‐color antibody surface marker expression panel to physical separate various putative populations of tip and stalk ESC‐derived EC (ESC‐EC) and replate as single cells. Some of the subpopulations of tip and stalk ESC‐EC were clonogenic with distinct proliferation rates, and morphological, functional, and genetic subphenotypes. Successful clonogenic tip and stalk ESC‐EC were isolated as Dll4+/VE‐cad+/Notch1‐ and Dll4‐/VE‐cad+/Notch1+, respectively. Moreover, greater numbers of tip cells (51%) were clonogenic compared with stalk cells (4%), suggesting that stalk cells need tip cells for survival and/or maintenance. Most importantly, these studies show that we can direct stem cells into specialized EC subphenotypes in vitro to address a range of therapeutic applications for these cells. Research funding is provided by CIRM grant Basic Biology V.
Characterization of Diabetic Cardiomyopathy: A Role for KDAC Activity
Authors: Waldman Maayan , Arad Michael , Leviner Dror , Aravot Dan , Laniado‐Schwartzman Michal , Abraham Nader , Hochhauser Edith
Published: June 2021

The molecular mechanisms leading to myocyte injury, hypertrophy and dysfunction in the diabetic heart are triggered by the characteristic metabolic abnormalities, and by abnormal energetics. Lysine (K) deacetylases (KDACs) inhibition was shown to be protective in models of doxorubicin cardiotoxicity through the reduction of fibrosis, decreased inflammation and the upregulation of antioxidant defenses. KDACs' role in the pathogenesis of diabetic heart disease is unknown.

We used a murine model of leptin resistance (db/db mice). These mice were treated for 1 month with angiotensin II to induce severe cardiomyopathy. Echocardiography heart morphology, biochemical markers of injury were measured in the serum and KDAC activity and signaling was measured in cardiac tissues by Western blot and real time qPCR.

Hypertensive diabetic mice developed a unique cardiomyopathy characterized by preserved fractional shortening (FS), thickened left ventricle wall and smaller ventricle cavity. Histological staining revealed infiltration of immune cells and extensive fibrosis. Biochemical markers indicated metabolic syndrome. Hypertrophic markers of diabetic hearts exhibited elevated BNP and MMP2 and lower KDAC activity. Elucidating the role of epigenetic mechanisms and KDAC in the development of diabetic cardiomyopathy in order to search for therapeutic strategies based on pharmacological manipulation of histone acetylation, is needed. Funded by Tel Aviv University.