bluuebunny/crossref_metadata_2025 huggingface dataset:
Hesperetin Inhibits Pro‐inflammatory Cytokine Secretion Induced by Lipopolysaccharide in Differentiated Human THP‐1 Monocytes under Hyperglycemic Conditions
Authors: Lee Aeri , Yun Jung‐Mi
Published: November 2023

Diabetes is a chronic disease characterized by high serum glucose levels. Chronic and acute hyperglycemia could be critical during the development of diabetic complications. Hesperetin is a type of bioflavonoid‐based flavanone found abundantly in citrus fruits. Hesperetin exhibits many pharmacological properties, including anti‐inflammatory, anticancer, and anti‐vascular formation effects. However, the epigenetic mechanisms mediating the antidiabetic and anti‐inflammation activities of hesperetin are still unknown. In this study, we investigated the effects of hesperetin on pro‐inflammatory cytokine secretion through the nuclear factor (NF)‐κB signaling pathway and the underlying epigenetic mechanisms in differentiated human THP‐1 monocytes treated with high glucose concentrations and LPS. THP‐1 cells were differentiated into macrophages by treatment with phorbol 12‐myristate 13‐acetate (PMA, 1 μM) for 48 h. Differentiated THP‐1 cells were cultured under normoglycemic (NG, 5.5 mmol glucose) or hyperglycemic (HG, 25 mmol glucose) conditions in the absence or presence of hesperetin (10, 25, 50, or 100 μM) for 48 h and then treated with 100 ng/mL LPS 6 h prior to harvesting. Under in vitro conditions mimicking diabetes complications, we found that hesperetin inhibited NF‐κB expression and acetyl p65 levels. Histone acetylases (HATs) and histone deacetylases (HDACs) have been found to play important roles in the regulation of several inflammatory and diabetic target genes by removing an acetyl group from lysine residues of target proteins. SIRT3 and SIRT6 modulate glucose and lipid metabolism. Thus, we examined HAT and HDAC activity using by ELISA. Hesperetin restored histone acetylase (HAT) and HDAC activities, similar to NG conditions, in high glucose‐ and LPS‐treated differentiated THP‐1 cells. High glucose and LPS stimulation increased NF‐κB, SIRT3 and SIRT6 levels and increased HDAC3 levels whereas hesperetin treatment reduced SIRT3 and SIRT6 expression and suppressed HDAC3 expression using western blotting, real‐time pcr and immunofluorescence. Accordingly, we concluded that HDAC3, SIRT3, and SIRT6 could be potent targets for the management of inflammation, infection, diabetes, and diabetic complications. Analysis of post‐transcriptional histone modifications showed that high glucose and LPS increased H3K9ac levels, whereas hesperetin suppressed H3K9ac levels. Based on these findings, we concluded that hesperetin may have applications in the treatment and prevention of diabetes and its complications.
GDF5 Is a Direct Target of LMX1B Regulation During Limb Dorsalization
Authors: Pira Charmaine U , Haro Endika , Pernu Lauren M , Casiano Carlos M , Kim Emily J , Aka Chelsea A , Oberg Kerby C
Published: November 2023

Lmx1b is a homeodomain transcription factor that is expressed in the dorsal mesoderm and plays a critical role in limb dorsalization. The absence of functional Lmx1b results in a ventral‐ventral limb phenotype in mice whereas ectopic Lmx1b expression causes dorsal‐dorsal limb morphology. Lmx1b‐regulated genes have been identified in developing limb buds by gene array analysis; however, direct targets of Lmx1b have yet to be confirmed. Lmx1b ChIP‐seq analysis of E12.5 mouse embryo limb tissue identified an Lmx1b bound interval (LBI) located 78Kb upstream of the Gdf5 gene, which is involved in joint development and structure and previously shown to be upregulated by Lmx1b at this stage of development. The LBI is highly conserved across divergent species and is associated with active chromatin regulatory marks supporting a role as a regulatory sequence. We isolated the LBI, cloned it into a GFP reporter construct and electroporated the construct into developing chick wings. We found that the LBI exhibited enhancer activity in limb joints, coincident with Gdf5 expression. Our findings support a recent report that identifies this cis‐regulatory module (CRM) as regulator of Gdf5 expression in distal joints. In addition, using chromosome conformation capture (3C) technology, we found that this CRM interacts directly with the Gdf5 promoter. Collectively, these data indicate that Lmx1b directly regulates Gdf5 via a joint‐restricted CRM during limb dorsalization. Further studies are warranted to characterize this Gdf5‐associated enhancer and the role Lmx1b and its partners play in regulating Gdf5 expression during limb dorsalization.
Oldenlandia diffusa Extracts Exert Anti‐Tumor Effects on Human Ovarian Cancer Cells Via an Epigenetic Regulator, KDM1B
Authors: Lee Yeon Kyu , Lim Jin Yeong , Joo Jong Cheon , Park Soo Jung , Park Yoon Jung
Published: November 2023

Ovarian cancer is the most fatal gynecologic malignancy among women with its high risk of late diagnosis and recurrence. Cisplatin is a platinum‐based anti‐tumor drug, which is widely used against ovarian cancer, however, recurrent tumors after the treatment frequently demonstrate acquired chemo‐resistance. Therefore, identification of alternative treatment for ovarian cancer seems critical. Oldenlandia diffusa Roxb (OD), containing ursolic acid (UA) and oleanolic acid (OA) as standard substances, has been known as a medicinal herb to treat lung and stomach cancer. However, the effect of OD on ovarian cancer is uncertain. This study was aimed to uncover the anti‐tumor effects of OD extracts and/or cisplatin on ovarian cancer cells, in particular carrying cisplatin‐resistance. OD was extracted by two ways using water and 50% methanol. As ovarian cancer cell models, cisplatin sensitive cell A2780 and the counterpart cisplatin resistant cell A2780cis were used. The methanol extract effectively reduced cell viability in both A2780 and A2780cis cells, while the water extract affects significantly cell viability in only A2780 cells, not in A2780cis. The effect of the methanol extract was greater than its standard substances, including UA and OA in both A2780 and A2780cis. Interestingly, combined treatment of cisplatin and the methanol extract substantially reduced cell viability of A2780cis cells, compared to single treatment of either cisplatin or the methanol treatment. Genome wide expression analysis using microarray data revealed that expression of an epigenetic marker lysine demethylase 1B (KDM1B) was upregulated in A2780cis cells compared to A2780 cells, and the result was confirmed by further quantitative analysis. Consistently, treatment of the methanol extract on A2780cis cells significantly reduced KDM1B mRNA expression in a dose‐dependent manner. Moreover, siRNA‐mediated knockdown of KDM1B reduced cell viability of A2780cis under cisplatin treatment, suggesting that KDM1B upregulation in A2780cis, in part, contributes to the cisplatin resistance. The results showed that the OD extract may promote cell death in the resistance cells under cisplatin through modulating KDM1B.
Which Handgrip Strength Cut‐offs Best Predict Low Physical Function in Older Adults?
Authors: Murphy Caoileann H , McMorrow Aoibheann M , Elroy Kate Mc , Cummins Helen , McGowan Maureen J , Rafferty Sheena , De Vito Giuseppe , McCarthy Sinead N , Corish Clare A , Egan Brendan , Roche Helen M
Published: November 2023

Introduction

The progressive loss of muscle mass and function with age (sarcopenia) and the age‐related loss of strength (dynapenia) are major causes of disability among the elderly1. Handgrip strength (HGS) is a quick and cost effective means of assessing muscle strength and is easily employed in clinical practice. The European Working Group on Sarcopenia in Older Persons has recommended two different sets of cut‐off points for the identification of low grip strength. One set are specific to sex only and the other set are specific to both sex and body mass index (BMI). However, it is currently unclear which set of cut‐off points are most useful for clinicians in predicting low physical function in older adults.

Aim

To determine if the sex/BMI‐based HGS cut‐offs are better at predicting poor physical function than the cut‐offs based on sex alone.

Methods

In a cross‐sectional analysis, anthropometry, HGS and physical performance were assessed in 407 community‐dwelling adults aged 65 years or over (mean(SD) age 77(8)y, BMI 27.9(4.5)kg/m2). HGS was measured via a handgrip dynamometer and physical performance by the Short Physical Performance Battery (SPPB). Linear and binary logistic regression were performed to examine which cut‐off points were better predictors of SPPB performance and multiple regression was used to assess the influence of age, sex and BMI on HGS.

Results

The percentage of subjects classified as dynapenic was 52.1% and 49.6% via the sex‐based and the sex/BMI‐based cut‐offs, respectively. Age and sex were significant predictors of HGS (β = −.42, p < .001 for age and β = .53, p < .001 for sex) but BMI was not (p > .05). The sex/BMI‐based cut‐offs explained a similar amount of variance in SPPB score compared to the sex‐based cut‐offs (24% vs. 23%). However subjects classified as having weak HGS by the sex‐based cut‐offs were more likely to be classified as having low SPPB performance than those classified as having weak HGS by the sex/BMI‐based cut offs (OR = 4.22, p < .001 for sex‐based cut‐offs vs. OR = 1.89, p = .07 for sex/BMI‐based cut‐offs).

Conclusion

BMI is not a significant predictor of HGS and, counter to our hypothesis, the sex‐based cut‐offs better predicted low physical performance compared with the sex/BMI‐based cut‐offs in a cohort of community‐dwelling older adults in Ireland.

Support or Funding Information

This work was supported by funding from the Department of Agriculture, Food and the Marine and Health Research Board through the Joint Programming Initiative ‐ A Healthy Diet for a Healthy Life (JPI HDHL) Knowledge Hub on Malnutrition in the Elderly (MaNuEL).
Dietary patterns associated with incidence of metabolic syndrome and its components
Authors: Perez‐Rodriguez Marcela , Ramirez‐Palacios Paula , Salmeron Jorge , Talavera Juan
Published: November 2023

Introduction

Metabolic syndrome (MetS) is a cluster of biochemical and physiological abnormalities associated with the development of cardiovascular disease and type 2 diabetes. MetS is present when three or more of the following five criteria are met: waist ≥ 102 cm in men and ≥ 88 cm in women, triglycerides ≥ 150 mg/dl, HDL‐C <40 mg/dl in men and < 50 mg/dl in women, systolic blood pressure ≥ 130 mmHg or diastolic blood pressure ≥ 85 mmHg and fasting glucose ≥ 110 mg/dl. Diet is one of the main risk factor for MetS and the study of regional dietary patterns is necessary to develop ad hoc recommendations for populations.

Objective

Study the effect of dietary patterns on the incidence of metabolic syndrome and its components.

Methods

We present a cohort study with a baseline measurement in 2004–06 and a follow up 6 years later. A total of 557 NW and 489 OW/OB healthy Mexican adults were included. Fasting glucose, TG, HDL‐C, WC and BP were measured, physical activity (PA) and diet data were self‐reported. Using principal components analysis (PCA), we identified dietary patterns. PCA provides a score for each dietary pattern for all participants; scores were classified in tertiles and adherence to each pattern was determined according to each pattern score tertile. Logistic regression analysis was performed with MetS and each of its components as dependent variables, including only subjects free of metabolic disease at the beginning of the study.

Results

Four dietary patterns were identified. There was no association between any dietary pattern and metabolic syndrome, waist circumference, triglycerides, HDL or blood pressure, nonetheless. After adjusting for relevant covariates, a high adherence to the dietary pattern with high loads for artificial sweeteners, wine and beer had a higher risk (OR=1.9 CI95% 1.1, 3.4) for altered glucose than a low adherence to that same pattern; and a high adherence to the dietary pattern with high loads for rice, corn and beans had a lower risk (OR=0.57 CI95% 0.33, 0.97) for altered glucose than a low adherence to that same pattern.

Conclusion

Traditional Mexican dietary pattern (rice, corn and beans) is associated with lower odds of developing altered glucose in a follow up period of 6 years.
Caloric Restriction Protects Against Hepatic Oncogenesis
Authors: Ploeger Jonathan , Mashek Douglas
Published: November 2023

In developed countries, obesity and its comorbidity non‐alcoholic fatty liver disease are well recognized risk factors for liver cancer, the sixth most common and second most deadly cancer globally. Currently, the only treatment option for hepatic cancer is liver resection or transplantation, procedures which are often contraindicated for patients with fatty liver disease. Caloric restriction (CR) has been shown in several animal models to extend disease free life span and reduce certain types of cancer. However, no studies have evaluated the effects of CR on hepatic cancer initiation. Here, we show for the first time, that restricting calories to 70% of ad libitum fed control groups (i.e. CR) provides robust protection from hepatic oncogenesis in response to the carcinogen diethylnitrosamine (DEN). As expected, CR completely prevented hepatic tumor formation and significantly reduced body weight gain and liver weights. Serum analysis showed no difference in free fatty acid concentration while CR mice tended to have increased ketone body concentrations suggesting increased fatty acid oxidation. Histological analyses performed by a blinded pathologist confirmed gross morphologic findings, demonstrating CR drastically reduced steatosis, hepatocyte ballooning, and immune cell infiltration. Additionally, pathological analysis of tumor and adjacent non‐transformed tissue confirmed CR completely ablated tumor formation and significantly reduced internal foci development. Immunohistochemical analysis of the proliferative marker Ki‐67 in non‐transformed tissue demonstrated CR greatly reduced positively stained nuclei compared to ad libitum fed mice. Collectively, our data demonstrate CR is a robust deterrent of hepatic oncogenesis and the co‐morbidities often attributed to liver cancer. Given the rise in incidence of HCC in developed countries, results from this study provide an important step in identifying translational strategies for therapeutic intervention and clinical prevention of liver cancer.

Support or Funding Information

This work is supported through an institutional grant generously provided by the “Healthy Foods Healthy Lives Institute” at the University of Minnesota
Eligibility of Rheumatoid Arthritis Patients for a Phase 1b Clinical Trial Evaluating Turmeric Supplementation
Authors: Strom Meghan , Alfafara Chelsea , DeSalvo Janel , McCallum Mia , Hopkins Laura , Pace Thaddeus , Chow Sherry , Levinsky Dale , Ruderman Eric , Billheimer Dean , Funk Janet
Published: November 2023

Rheumatoid arthritis (RA) is an autoimmune disease that primarily targets the joints, leading to synovial inflammation. Turmeric supplementation markedly reduces joint inflammation in pre‐clinical models of RA, suggesting a potential benefit in reducing joint inflammation in patients with RA. Here, we evaluate the percentage of patients eligible for a randomized controlled trial (RCT) of turmeric dietary supplements in RA, evaluating tolerability and pharmacokinetics. Recruitment materials were distributed in both rheumatology offices and community settings. Study entry eligibility criteria included having active disease (presence of inflamed and/or swollen joints), no current biologic use, as well as other health and medication criteria. Individuals were screened for basic eligibility through a telephone interview followed by a more comprehensive in‐clinic screening. In total, 200 patients were referred between September 2015 through October 2016; the majority (85%, n= 170) were self‐referred from community‐based recruitment strategies. From a pool of 171 (86%) who completed telephone screening, 40% (n=68) were disqualified from participation due to geographical location or having no clinical diagnosis of RA. Of the remaining 103 referrals, only 4% of local RA patients (n= 4) met all eligibility criteria; 15 (15%) discontinued the screening process and 84 (82%) were ineligible. Primary reasons for ineligibility included current or failed biologics (37%, n= 31) and no active disease (18%, n= 15). Other reasons included a serious comorbid medical condition (14%, n=12), unstable medication doses (13%, n= 11), high non‐steroidal anti‐inflammatory use (12%, n=10) or a history of malignancy (6%, n= 5). A greater proportion of patients referred from rheumatology practices passed telephone screening than those self‐referred from the community (20%, n= 6 vs. 8%, n= 14). Few of the patients screened met all eligibility requirements. Although uncommon, some patients did not wish to participate in the trial due to the possibility of receiving a placebo, as well as having to discontinue current turmeric supplement use. In summary, recruitment strategies that are primarily community‐based are inefficient in enrolling subjects who meet eligibility criteria for a RCT of turmeric dietary supplements in RA.

Support or Funding Information

NIH (NCCIH): R34AT007837‐03
Inhibition of H1299 Human Lung Cancer Cell Proliferation and Survival by Carnosic Acid is Associated with Akt Inhibition and AMPK Activation.
Authors: Tsiani Evangelia Litsa , Moore Jessy
Published: November 2023

Non‐small cell lung cancer (NSCLC) represents a very aggressive form of lung cancer and is characterized by resistance to chemo‐ and radiotherapy thus, finding new ways to inhibit NSCLC cell proliferation and survival is very important. Carnosic acid (CA) is a phytochemical found in rosemary, a frequently used herb which is regarded as safe for consumption. Phytochemicals represent an important opportunity to explore new agents with chemotherapeutic potential and to understand their novel and potentially relevant mechanisms of action. In vitro and in vivo evidence exists supporting anticancer effects of CA however, no studies have explored the role of CA in H1299 NSCLC cells and much is still unknown about its mechanism of action. The objectives of the present study were to examine the effects of CA on H1299 cell proliferation and survival, and to investigate its effects on the phosphatidylinositol 3‐kinase (PI3K)‐Akt, AMP‐activated protein kinase (AMPK) and extracellular signal‐regulated kinase (ERK) signaling pathways which regulate cell growth and survival. Cell proliferation was measured by crystal violet assay, clonogenic assays were performed to examine the effects of CA on cell survival, and immunoblotting with phospho‐specific antibodies was performed to examine signaling events. CA dose‐dependently inhibited H1299 cell proliferation. Significant inhibition of cell proliferation and survival was seen with concentrations of 25 and 50μM CA. At 50μM CA inhibited Akt activity and enhanced ERK and AMPK activity in H1299 cells, without affecting total levels of each protein. Our data suggest that CA may have potent anticancer properties in NSCLC and warrants further investigation of its use as a chemotherapeutic.

Support or Funding Information

Brock University Advancement Fund (BUAF) grant to ET.
Natural Product Supplement Use for Rheumatoid Arthritis Self‐Management in the United States: A Literature Review
Authors: DeSalvo Janel , Strom Meghan , Haiber Karen , Feng Lena , Hopkins Allison , Funk Janet
Published: November 2023

Natural‐product dietary supplements (NDS) are a frequently used complementary and alternative medicine (CAM) modality in the United States (U.S.), with complaints of musculoskeletal symptoms being a primary reason for use. Rheumatoid arthritis (RA) is a less common, but particularly aggressive, inflammatory form of arthritis where pharmaceutical treatment is associated with potential toxicities and high cost. NDS use for the purpose of reducing symptoms could be associated with higher risks of supplement‐drug interactions in this population. To examine patterns of NDS use for self‐management of RA in the U.S., a comprehensive literature search was carried out using the Medline database; bibliographies were also reviewed to identify additional articles. Key words used included: botanicals, osteoarthritis, arthritis, rheumatoid arthritis, RA, supplements, complementary, herbal, alternative, and CAM. Included articles were those with ≥ 50% RA patients that evaluated NDS use. In total, 23 articles were identified, of which 4 pertained to patients in the US (California, New York, North Carolina and Alabama). CAM use was highly prevalent in all US studies, however the types of CAM varied widely across studies and included ointments, special diets, and nutritional supplements. Special diets (e.g. vegan, arthritis diet) were among the least commonly used forms of CAM in all but one study. NDS use was reported by 30–52% of RA patients. Despite temporal and regional differences across surveys, including disparate or undocumented demographics, particular supplements were consistently reported. Glucosamine and chondroitin were included in the top 4 most commonly used NDS in 3 of the 4 surveys. Fish oil, cod liver oil, garlic, and vinegar/honey were among the top supplements used in more than one survey. Other frequently used supplements appearing in a single survey included green tea, gingko biloba, alfalfa and yucca. Common reasons for use included perceived helplessness, disease manifestations, duration of illness and failure of biomedical treatments. The prevalence of NDS use appears to be higher in RA patients than in the general U.S. population. Still, specific use by individuals with RA is not well documented, and in many regions of the U.S., remains largely unknown. Because of the potential for supplement‐drug interactions in this population due to intensive treatment with pharmacologic agents, elucidation of additional information about specific NDS use by RA patients should be an important focus in both clinical practice and future research.
Effect of an educational group intervention in self‐care behavior for patients with type 2 diabetes on HbA1c and postprandial capilar glucose
Authors: Castillo Judith Juarez , Avendaño Carlos , Carriedo Alicia Parra , Izeta Ericka Ileana Escalante
Published: November 2023

Type 2 Diabetes Mellitus (T2DM) is a chronic, degenerative and progressive condition that includes a group of metabolic diseases characterized by hyperglycemia secondary to a partial or total defect of insulin secretion or action. In Mexico, T2DM represents a disabling disease at a public health problem level due to the micro and macrovascular complications associated with the disease and poor glycemic control (HbA1c). The Health Center “Centro Graber” in Hidalgo, Mexico has a group of patients (n=20, 26 – 72 years old) living with T2DM in which some educational needs in self‐care behaviors were observed.

Aim of study

Assess the benefits of an educational program in self‐care behaviors in HbA1c and postprandial capilar glucose.

Methods

HbA1c (boronate fluorescence quencing technology) and postprandial capilar glucose values were measured at “Centro Graber”. Adherence was assessed by the attendance of the subjects.

Results

Attendance of patients was high (99.33%). Significant and clinically relevant changes in HbA1c (−1.42%, p=0.039) and postprandial capilar glucose (−28.67 mg/dl, p=0.058) were observed.

Conclusions

Group educational programs in self‐care behaviors may be an effective strategy when combined with medical treatment for improvements in glycemic control (HbA1c) and postprandial capilar glucose in patients with T2DM who attend the Health Center “Centro Graber”.
Determining the Association of Youth's Physical Activity and Sedentary Behaviors with Weight Status
Authors: Taylor Erin M , Keim Kathryn S. , Mullen Mary C. , Sowa Diane C. , Lui Karen J.
Published: November 2023

Introduction

Over the past decade the weight status of children in the United States has been increasing, making it an urgent health problem. Physical activity and screen time behaviors have been associated with the prevention and treatment of childhood obesity. Recommendations for these behaviors were set by the Expert Committee in order to prevent and treat overweight and obesity.

Objective

To determine the association between the youth's demographic, physical activity, and sedentary behaviors with weight‐status using items assessed on the Child Nutrition and Physical Activity (CNPA) screener.

Design

A cross sectional study was conducted using a retrospective chart review method.

Methods

The CNPA screener was given to all parents who came to the Rush University Pediatric Primary Clinic between 2012–2015. Responses to questions regarding physical activity, sedentary behavior, and parent perception of youth's weight status were assessed. Chi square analysis was used to determine the association between demographic characteristic, physical activity, sedentary behaviors, weight status, and weight status perception. Forward Wald logistic regression analysis was conducted with odds of being overweight, obese, and severe obese compared to healthy weight. All procedures were approved by IRB.

Results

The sample consisted of 2,230, 2–18 year old children who were 51% male, 60% non‐Hispanic Black, and 61% of a healthy BMI percentile. Using forward Wald logistic regression analysis, 9 variables were entered into the model‐age, sex, race, meeting physical activity recommendation, meeting media recommendation, whether a TV was in the bedroom, and accuracy of weight perception. The model was statistically significant χ2 (6, N=2079) = 1093.37, p <.001, indicating that the model was able to distinguish between healthy weight children and those who were overweight, obese, or severe obese. The model explained 40.9% to 55.5% of the variance in weight status and correctly classified 83.5% of cases. Increasing age (1.14 OR (95% CI 1.10–1.17)) (p<.001) and being Hispanic 1.72 OR (95% CI 1.30–2.26) (p<.001) increased the odds of being overweight, obese, or severe obese. Meeting physical activity recommendation (0.60 OR (95% CI 0.45–0.81)) (p=.001) media recommendation 0.76 OR (95% CI 0.57–0.996) (p=.047), not having a TV in the bedroom (0.67 OR (95% CI 0.51–0.87)) (p=.002), and parent accurate weight perception (0.02 OR (95% CI 0.01–0.03)) (p<.001) decreased the odds of being overweight, obese, or severe obese.

Conclusion

The odds of being overweight or obese were higher if the child was older or Hispanic and lower if they met the physical activity, media, no TV in bedroom recommendations and had accurate perception of weight status. Meeting the recommendation for all physical activity and sedentary behaviors entered in to the model decreased the odds of being overweight or obese.

Support or Funding Information

Departmental funding
Novel vitamin D‐Nanoemulsion improves testicular function of vitamin D deficient rats
Authors: Alruwaili Nawaf Wanas , Abulmeaty Mahmoud M.A. , Almajwal Ali M , Elsadek Mohammed F , Razak Suhail , Andrade J.E.
Published: November 2023

Background and aim

Vitamin D (VD) deficiency is associated with disturbed reproductive functions in male rats which might be ameliorated by vitamin D (VD) supplementation. In this study, the efficacy of a novel VD carrier, developed by sonication and pH‐shifting of pea protein isolate (nanoemulsion), in improving testicular function of VD‐deficient rats was investigated.

Methods

Thirty weaned male albino rats (92.62±13.61g) were divided into 2 groups. Group I (n=10) was fed a synthetic AIN‐93G diet while Group II (n=20) was fed a customized AIN‐93G diet with carbohydrate 59.3%, protein 18.1%, fat 7.1%, fiber 4.8%, VD (0–50 IU/kg) and calcium (5.1 g/kg) for 6 weeks. Hypovitaminosis D was confirmed by taking blood samples from the lateral tail vein. Rats in group I were used as a sufficient control (SC group), while those of group II were further subdivided into two groups (n=10 each): a nanoemulsion without VD deficient control (DC group) and a nanoemulsion with VD treatment (DT group). Animals were treated for 2 weeks with a gavage dose (1 mL of 27 μg/mL) every other day. Body weights were measured daily. Plasma 25‐hydroxy vitamin D (25OHVD) and testosterone (T) were measured by ELISA. After euthanasia, testis were isolated, sectioned, stained and examined under light microscope. Image J was used to analyze the selected regions of interest (ROIs).

Results

VD deficient diets resulted in low serum 25OHVD levels in SC vs. deficient groups (36.8±9.1 vs. 15.9± 5.8 μg/mL, p<0.001), but no changes in body weight. VD deficient rats showed marked reduction of T (DC vs. SC, 0.83±0.22 vs. 3.06±0.74 ng/mL, p<0.001). Serum 25OHVD was not significantly correlated with T levels (r=0.408, p=0.18). Histology showed a significant reduction in the Leydig cell mass and degenerative changes in all levels of the spermatogenesis (Figure a and b). After treatment, DT group showed 6‐fold increase in serum T in comparison with that of DC group and double that of SC (7.30±1.18 vs. 0.83±022 and 3.06±0.74 ng/mL respectively, p<0.0001 for all). Microscopically, selected ROIs showed marked improvement of germinal epithelium and all stages of spermatogenesis, together with an increase in Leydig cell count.

Conclusions

Oral VD supplementation using pea protein nanoemulsion could improve serum T levels and ameliorate VD deficiency‐induced histopathological changes in testis. Future research should investigate the underlying mechanisms.

Support or Funding Information

This project is funded by the national plan of science, technology and innovation, King Abdul‐Aziz City for Science & Technology (KACST) project no: 12‐NAN 2576‐02
Effect of Enteral Docosahexaenoic Acid on Retinopathy of Prematurity During Their Hospital Stay
Authors: Bernabe‐Garcia Mariela , Dominguez‐Vallejo Paola , Cruz‐Reynoso Leonardo , Villavicencio‐Torres Astrid , Villegas‐Silva Raul , Inda‐Icaza Patricia
Published: November 2023

INTRODUCTION

The retinopathy of prematurity (ROP) is a proliferative vasculopaty of the retina; in 2001, the WHO estimated near of 50,000 children in the world with childhood blindness was related with. In Mexico is the first preventable cause. There is no effective treatment for ROP. The docosahexaenoic acid (DHA) has demonstrated to reduce inflammation, neovascularization and obliteration of blood vessels of the retina in experimental models4, and in 2 studies using parenteral nutrition (PN) lipid emulsion. However, PN may cause complications such as colestasis and sepsis catheter‐related. To our knowledge, there is no study that evaluates enteral DHA on ROP.

OBJECTIVE

To evaluate the effect of enteral DHA on development of retinopathy of prematurity and their severity during hospital stay of preterm infants.

METODOLOGY

Double‐blind, clinical trial. Preterm neonates were recruited with birth weight <1500g but equal or higher than 1000g DHA group received 75 mg of DHA/kg/day (Life's DHA, Martek Life Enriched®) and control group sunflower oil, throughout 14 days by enteral feeding. They are blinded as A and B codes. We did an interim analyses on ROP incidence and severe ROP (stage 3 with or without plus, or other stage with plus). Groups were compared with relative risk (RR), Fisher's Exact, T‐Student or U‐Mann‐Whitney tests, as appropriate. Logistic regression was applied to adjust by confounders. Clinical‐Trial ID: NCT02683317

RESULTS

We recruited 90 patients, 45 from each group. There were no differences between groups in total ROP incidence (36% vs. 42%, p= 0.333) nor its RR=0.842 (95%IC 0.500–1.418). Likewise on severe ROP incidence (81% vs. 58%, p= 0.132) or its RR= 0.445 (95%IC 0.141–1.404). Interestingly, A group shown a borderline reduction in the probability of severe ROP after adjust by NSAIDs, steroids, gestational age at birth, duration with tissue saturation higher than 95%, events of apnea, tissue DHA content in erythrocytes and presence of respiratory distress syndrome (OR adj= 0.05; 95%IC 0.002–1.127; p=0.060; R2 model=61.2%).

CONCLUSION

The preliminary results suggest that enteral DHA at dose of 75 mg/kg/day might have a prophylactic effect on the incidence of severe ROP.

Support or Funding Information

This project received a grant from CONACYT SALUD‐2015‐2‐261765
Regulation of folate‐mediated one‐carbon metabolic kinetics by glycine supply
Authors: Chiang EnPei Isabel , Huang Yu‐Hsuan
Published: November 2023

Glycine‐N methyltransferase (GNMT) is a major hepatic enzyme that converts S‐adenosylmethionine to S‐adenosylhomocysteine while generating sarcosine from glycine. The metabolic role of GNMT in folate‐mediated one‐carbon metabolism and how glycine supply may affect one carbon metabolic kinetics are under investigated in the present study. Hepatocyte‐derived cell‐lines with and without GNMT expression were cultured in minimum essential medium (MEM) with and without high glycine supply. Methylene‐tetrahydrofolate (mTHF) utilization and endogenous formate biosynthesis were carefully investigated using stable isotopic tracers and GC‐MS. We discovered that high glycine inhibits the M+1 specie enrichments in deoxythymidine (dT+1) from 2,3,3D3‐serine, suggesting that high glycine may inhibit endogenous formate production in these in vitro models. In a parallel experiment that exogenous formate was used as the 1C source, enrichments in dT+1 significantly increased from exogenous formate, supporting our hypothesis that high glycine inhibits endogenous formate production for thymidine synthesis. Mice fed in amino acid based diet supplemented with high glycine had significantly lower plasma formate concentrations that provide in vivo evidence on our hypothesis. On the other hand, high glycine promotes M+1 specie enrichments in methionine from 2,3,3D3‐serine, indicating that high glycine supply may favor the utilization of methyleneTHF derived one carbon for homocysteine remethylation. High glycine supply can promote folate dependent methionine synthesis in our cell models. Preliminary in vivo experiments demonstrated that high dietary glycine alters folate dependent methionine synthesis, consistent with our in vitro findings. We demonstrated how glycine supply regulates folate‐mediated one‐carbon metabolic kinetics in vitro and in vivo.

Support or Funding Information

MOST 105‐2320‐B005‐010‐MY3; MOST104‐2911‐I005‐301; and Ministry of Education, Taiwan, R.O.C. under the ATU plan)
Geranylgeraniol Suppresses the Expression of Adipogenic Genes and the Differentiation of Murine 3T3‐F442A and 3T3‐L1 Preadipocytes
Authors: Elfakhani Manal , Yount Sophie , Mo Huanbiao
Published: November 2023

Fundamental to the pathology of obesity is the hypertrophy and hyperplasia of adipocytes. Lipid accumulation in differentiated adipocytes is part of the adipocyte lifecycle that eventually leads to recruitment of macrophages and inflammation. We have previously shown that lovastatin, a competitive inhibitor of 3‐hydroxy‐3‐methylglutaryl coenzyme A (HMG CoA) reductase, suppresses the differentiation of murine 3T3‐F442A adipocytes via mevalonate deprivation. Tocotrienols, down‐regulators of HMG CoA reductase, inhibit adipocyte differentiation via suppression of peroxisome proliferator‐activated receptor γ (PPARγ). We hypothesize that geranylgeraniol, a diterpene known to accelerate the degradation of HMG CoA reductase, mimics the impact of lovastatin and tocotrienol in suppressing adipocyte differentiation and the expression of adipogenic genes. Oil Red O staining and Adipo‐Red assay showed that a 7‐d incubation with 2.5 – 20 μmol/L geranylgeraniol decreased the intracellular triglyceride content of murine 3T3‐F442A and 3T3‐L1 adipocytes in a concentration‐dependent manner. Geranylgeraniol also down‐regulated the expression of PPARγ, a key regulator of adipocyte differentiation, as well as the expression of adipocyte marker genes comprising adiponectin, leptin, fatty acid binding protein 4 (FABP4) and lipoprotein lipase, as measured by real‐time qPCR. Concurrently, Western‐blot showed that geranylgeraniol reduced the levels of PPARγ, FABP4 and C/EBPα proteins. No cytotoxicity was observed in preadipocyte cells incubated with geranylgeraniol (0–400 μmol/L) for 24 and 48 h. Mevalonate‐derived metabolites have crucial roles in promoting adipocyte differentiation by regulating the expression of adipogenic genes. Dietary mevalonate suppressors as anti‐adipogenesis compounds may have potential in the prevention of obesity.

Support or Funding Information

American River Nutrition, Inc.
Effects of Folic Acid and 5‐Methyltetrahydrofolate Supplementation on Folate Metabolism and One‐Carbon Transfer Reactions in a Mouse Model
Authors: Im David , Bloemen Nicolaas , Sohn Kyoung‐Jin , Croxford Ruth , Kim Young‐In
Published: November 2023

Because of potential adverse health effects of folic acid (FA) supplementation, 5‐methyltetrahydrofolate (5MTHF) has been proposed as a safe alternative to provide supplemental levels of folate. However, the effects of 5MTHF versus FA supplementation on folate metabolism and one‐carbon transfer reactions have not been well studied in an in vivo model. We compared the supplemental effects of FA and 5‐MTHF on the expression of genes involved in intracellular folate metabolism and one‐carbon transfer reactions in the liver using a mouse model. Postweaning C57BL/6 mice were randomized to receive diets containing 2, 10, 20 mg FA/kg of diet or their molar equivalent 5MTHF diets for 12 weeks. Gene expression in the liver was assessed using real‐time RT‐qPCR. Plasma and hepatic folate concentrations accurately reflected supplemental levels of both FA and 5MTHF (p‐trend<0.0001). Expression of thymidylate synthase (Tyms) in the liver was not significantly different between FA and 5MTHFR at each molar equivalent supplemental levels. However, 20 mg 5MTHF supplementation was associated with a significant downregulation of Tyms compared with 10 mg 5MTHF (p<0.05). Expression of dihydrofolate reductase (Dhfr) was significantly higher with 5MTHF supplementation compared with FA supplementation at each molar equivalent concentration (p<0.0001). Expression of methylenetetrahydrofolate reductase (Mthfr) was significantly higher in the 10 mg 5MTHF group relative to its molar equivalent FA group (p<0.001), and relative to the 20 mg 5MTHF group (p<0.05). Expression of methionine synthase (Mtr) was significantly greater in all 5MTHF groups relative to their molar equivalent FA groups (p<0.001). Expression of DNA methyltransferase 1 (Dnmt1) was not significantly different between the 5MTHF and FA groups at each molar equivalent concentration. However, Dnmt1 expression was greater in the 10 mg FA and 5‐MTHF group relative to the 2 mg FA group and 20 mg 5‐MTHF group, respectively (p<0.05). Expression of Dnmt3a was greater in all 5MTHF groups relative to their molar equivalent FA groups (p<0.05), and was significantly greater in the 20 mg 5MTHF group relative to the 2 mg 5MTHF group (p<0.05). Expression of Dnmt3b was greater in the 10 and 20 mg 5MTHF groups relative to their equimolar FA groups. Our results suggest that 5MTHF supplementation elicits differential effects on the expression of genes involved in folate metabolism and one‐carbon transfer reactions compared with FA supplementation. Studies are underway to determine functional ramifications associated the observed differential gene expression between 5MTHF and FA supplementation.
Alpha‐Lipoic Acid Inhibits Expression of Monocyte Chemoattractant Protein‐1 in Helicobacter pylori‐Infected Gastric Epithelial Cells
Authors: Kyung Seo Yeon , Lim Joo Weon , Kim Hyeyoung
Published: November 2023

Helicobacter pylori (H. pylori) causes gastritis and gastric cancers. Adenosine monophosphate‐activated protein kinase (AMPK) is a crucial regulator of energy metabolic homeostasis and a major survival factor in a variety of metabolic stresses. AMPK activation is related to anti‐inflammatory properties via inhibition of NADPH oxidase activity in various cells. Alpha‐lipoic acid (Alpha‐LA), a naturally occurring thiol compound, shows antioxidant and anti‐inflammatory effects. The aim of this study is to investigate whether alpha‐LA inhibits H. pylori‐induced expression of pro‐inflammatory cytokine monocyte chemoattractant protein‐1 (MCP‐1) via activation of AMPK in human gastric epithelial AGS cells.

Human gastric epithelial AGS cells were infected with H. pylori (strain NCTC11637) at bacterium/cell ratio of 100:1. mRNA expression of MCP‐1 were determined by real‐time PCR analysis while the levels of phospho‐AMPK and AMPK were determined by Western blot analysis. Levels of reactive oxygen species (ROS) was determined using DCF fluorescence. NADPH oxidase activity was measured by lucigenin assay.

As a result, H. pylori increased NADPH oxidase activity, ROS levels and MCP‐1 expression, but decreased AMPK phosphorylation in AGS cells. Alpha‐LA inhibited H. pylori‐induced increase in NADPH oxidase activity, MCP‐1 expression, and ROS levels in AGS cells. Alpha‐LA prevented H. pylori‐induced decrease in AMPK phosphorylation in AGS cells. In addition, AMPK inhibitor compound C suppressed inhibitory effects of alpha‐LA on H. pylori‐induced increase in NADPH oxidase activity, ROS production, and MCP‐1 level in AGS cells.

In conclusion, alpha‐LA may inhibit MCP‐1 expression by activating AMPK and inhibiting NADPH oxidase in H. pylori‐infected AGS cells. Consumption of alpha‐LA‐rich foods may prevent H. pylori‐associated gastric inflammation.
Effect of Leaf Extracts of Dendropanax morbifera on Selected Probiotics and Pathogenic Bacteria
Authors: Lee Yoo‐Hyun , Lee Jeong Yoon , Kim Ji Won , An Yeon Ju
Published: November 2023

Objectives

The objective of this study was to clarify any possible interaction between probiotics and leaf extracts of Dendropanax morbifera as prebiotics.

Methods

The leaf extracts of Dendropanax morbifera was prepared using various concentration of ethanol (0, 20, 50%). Bacterial growth was investigated in each probiotic (Lactobacillus acidophilus ATCC 4356, Lactobacillus leuteri ATCC 14485, Bifidobacterium bifidum ATCC 29521, Streptococcus thermophiles ATCC 14485) containing 1% (w/v) leaf extracts of Dendropanax morbifera. Total phenolic and flavonoid contents were determined and then, ABTS and DPPH assay were performed for evaluating radical scavenging activity in Dendropanax morbifera. Antimicrobial activity was measured against pathogenic bacteria (Listeria monocytogenes ATCC 19111, Clostridium difficile ATCC 9689, Staphylococcus aureus ATCC 6538p)

Results

In the growth curve, the decrease at the death phase was slower Lactobacillus acidophilus that contained aqueous extracts of Dendropanax morbifera leaf, Bifidobacterium bifidum and Lactobacillus leuteri that contained 50% ethanol extract, and Streptococcus thermophiles that contained 20% ethanol extract compared to the control. Especially, 50% ethanol extract showed the highest antioxidant capacity in ABTS assay. Also, the highest phenolic and flavonoid contents was observed in 50% ethanol extract. 50% ethanol extract exerts antibacterial activity against Listeria monocytogenes and Staphylococcus aureus.

Conclusion

These results demonstrate the leaf extracts of Dendropanax morbifera is a potent prebiotic, which leads to slower death of probiotics and exert antibacterial activity against pathogenic bacteria.

Support or Funding Information

This study was supported by a grant from HBJ Biopharm.
Effect of Folate Status on High‐Fat Diet‐Induced Obese Mice with Chronic Aristolochic Acid Nephropathy
Authors: Lin Bi‐Fong , Koh Xiang‐Lin , Wu Chi‐Heng , Huang Hsun‐Yi
Published: November 2023

A negative correlation has been reported between human serum folate levels and chronic kidney failure. Reducing folate intake in modern diets high in fat/calories along with low vegetables/fruits might be associated with increased risk for kidney disease in industrialized countries without folic acid fortification policy. To investigate whether folate status affects renal function in high‐fat diet‐induced obesity, four groups of C57BL/6 male mice were either fed normal‐fat diet with folate (NF‐f1) as negative control, or high‐fat diet without folate (HF‐f0), with folate (HF‐f1), with 10‐fold folate (HF‐f10). After 48 weeks' feeding, the HF groups obese mice were supplemented with 8 mg/Kg/day aristolochic acid (AA), a nitrophenanthrene carboxylic acid found in Aristolochic species used in the practice of traditional Chinese medicine, for 3 weeks to induce AA nephropathy (AAN) before sacrifice. The results showed that urine protein of mice fed AA‐containing diets, except the HF‐f10 group, significantly increased. Serum and renal galactin‐3, renal IL‐6 and TNF‐a in obese mice with chronic AAN were significantly higher when compared to the NF‐f1 mice without AA induction. The HF‐f10 mice had lower serum creatinine and higher IL‐10 secretion from ConA‐stimulated splenocytes. In contrast, the HF‐f0 mice had significantly higher urine protein, serum galaction‐3 and IL‐6, renal TGF‐b, and IL‐2 and TGF‐b secretion from splenocytes. Serum folate levels were negatively correlated with renal IL‐6, IL‐1b, TNF‐a, and IL‐10 in the HF‐f0 mice. Our study suggested that folate deficiency might promote, but folate supplementation might protect from renal inflammation and fibrosis progression in high‐fat diet‐induced obese mice.

Support or Funding Information

Funded by the Ministry of Science and Technology Science Council of the Republic of China [MOST105‐2320‐B‐002‐028‐MY3]
Biological Evaluation of a Novel Red Maple Leaf Extract for Nutraceutical Applications
Authors: Ma Hang , Liu Weixi , Dain Joel A. , Seeram Navindra P.
Published: November 2023

The red maple (Acer rubrum) and sugar maple (Acer saccharum) species, which are endemic to eastern North America, are widely known for their sap which is used produce the natural sweetener, maple syrup. Interestingly, various parts of the red maple tree have been used as traditional medicines by the indigenous people of North America and our laboratory has identified a series of glucitol‐core containing gallotannins (GCGs) from its aerial parts. These GCGs show a wide range of biological activities including antioxidant, anti‐diabetic, anti‐glycation, and anti‐proliferative effects. Herein, we report the biological evaluation of a red maple leaf extract (MLX) for functional food/nutraceutical applications. The MLX was evaluated in a panel of in vitro assays antioxidant, anti‐glycation, and anti‐acetylcholinesterase effects. At concentrations ranging from 20 to 500 μg/mL, MLX scavenged free radicals in the DPPH assay, as well as displayed ferric reducing antioxidant power in the FRAP assay. MLX showed inhibitory effects against the formation of advanced glycation endproducts and acetylcholinesterase enzyme with IC50 values of 157.6 and 327.3 μg/mL, respectively. In summary, this study suggests that MLX should be further investigated for functional food/beverage applications given its GCGs content and biological activities. Future studies in our laboratory will include the evaluation of MLX in animal models.