bluuebunny/crossref_metadata_2025 huggingface dataset:
Gut Microbiota Restores Central Neuropeptide Deficits Present in Germ Free Animals
Authors: Covasa Mihai , Duca Frank A , Swartz Timothy D
Published: November 2023

Recent work has shown that gut microbiota plays an important role in host energy homeostasis. Indeed, germ‐free (GF) mice have lower levels of adiposity and leptin levels compared to conventionalized mice. Additionally, GF mice are resistant to obesity when placed on a high‐fat diet. Studies show that gut microbiota contributes to obesity by increasing energy extraction, promoting inflammation, and altering lipogenic and adipogenic enzymes. Moreover, recent work demonstrates the ability of the microbiota to alter expression and release of gut peptides that control appetite and regulate energy homeostasis. However, little is known about the neuronal response of these hormones in GF animals, especially leptin, which is strikingly decreased in these animals. Therefore, we aimed to determine the sensitivity of leptin in GF mice compared to conventional mice by injecting leptin and measuring levels of leptin receptor activation via STAT3 and PI3K pathways in the hypothalamus and hindbrain. We also measured expression of orexigenic and anorexigenic peptides in the hypothalamus and hindbrain to examine whether gut microbiota impacts central nervous system regulation of energy homeostasis. We found that GF mice had a significant increase in hypothyalamic NPY and AgRP mRNA expression and a decrease in hindbrain NPY and AgRP mRNA. mRNA expression of POMC and CART remained unchanged. Protein expression of hypothalamic STAT3 and pERK/ERK increased significantly in response to peripheral leptin. Further, administration of leptin normalized circulating levels of leptin, GLP‐1, PYY and ghrelin, all of which were significantly decreased in GF mice. Conventionalization of GF mice (10 days) restored the deficits in hypothalamic and hindbrain neuropeptides present in GF animals. These results show that gut microbiota regulates hypothalamic and hindbrain orexigenic/anorexigenic neuropeptide expression. This is in line with the role of gut microbiota in lipid metabolism and fat deposition that may contribute to excess fat in conventionalized animals under high feeding condition.

Support or Funding Information

Romanian National Program PN‐II‐ID‐PCE‐2012‐4‐0608 no. 48/02.09.2013, “Analysis of novel risk factors influencing control of food intake and regulation of body weight”
Effects of Alcohol Consumption in Prevention of Colon Cancer in Rats
Authors: Klarich DawnKylee S , Penprase Jerrold , Cintora Patricia , Medrano Octavio , Erwin Danielle , Brasser Susan M , Hong Mee Young
Published: November 2023

Alcohol consumption is a risk factor associated with colorectal cancer (CRC); however, some studies have reported that moderate alcohol may not contribute additional risk for developing CRC while others suggest that moderate alcohol provides a protective effect that reduces CRC risk. The purpose of this study was to determine the effects of moderate alcohol (20% ethanol) administered on alternate days for 3 months in male Wistar rats on biomarkers that were associated with CRC development. Blood samples and colon mucosa samples were collected from the experimental alcohol and the control groups (n = 12/group) at the end of the study and were used to evaluate a variety of biomarkers that were associated with inflammation, DNA damage, antioxidant activity, and acetaldehyde clearance. Blood alcohol content, liver function enzyme activities, and 8‐oxo‐deoxyguanosine (8‐oxo‐dG) DNA adducts were also assessed in serum. Rats treated with moderate alcohol were found to have significantly lower 8‐oxo‐dG levels in blood, a marker of DNA damage, with no significant increase in repair activity by colonic 8‐oxoguanine DNA glycosylase 1 (OGG‐1), a base excision repair enzyme responsible for removing 8‐oxo‐dG damage. The blood alcohol content and liver enzyme tests were not significantly different between groups, with the exceptions of alanine aminotransferase and lactate dehydrogenase, which were both significantly lower in the alcohol group. Moderate alcohol significantly decreased cyclooxygenease‐2 gene expression, a marker for inflammation that is associated with CRC risk. The alcohol group had significantly increased glutathione‐S‐transferase M1 (GSTM1) expression, which is an antioxidant defense enzyme that helps detoxify carcinogens, like acetaldehyde, and also resulted in significantly increased aldehyde dehydrogenase 2 (ALDH2) expression, which allowed for greater acetaldehyde clearance. Increased expression of GSTM1 and ALDH2 likely contributed to reduce mucosal damage that is caused by acetaldehyde accumulation. Additionally, by assessing the correlations within the alcohol group we did find a positive relationship between increased alcohol intake and increased OGG‐1 and ALDH2 gene expression. Other positive correlations were found that provided evidence that increased 8‐oxo‐dG level was positively correlated with liver function enzyme activities and gene expression of RelA, OGG‐1 repair enzyme, and GSTM1 detoxification enzyme. These results suggest that moderate alcohol does not increase CRC risk and may provide beneficial effects that reduce CRC risk, which was evidenced by reduced inflammation activity and lower DNA damage after alcohol exposure.

Support or Funding Information

This study was funded by NIH AA023291. Additional funds were received from the Kasch‐Boyer Endowed Scholarship in Exercise and Nutritional Sciences.
Nutritional Intervention Improves the Total Energy Intake in Cancer Patients
Authors: Hung Shy‐Fen , Wen Cheng‐Fan , Li Pei‐Rong
Published: November 2023

Background

20% causes of death in cancer patients were malnutrition rather than the tumor itself. Cancer patients during the treatment period often occurs gastrointestinal symptoms, including nausea, vomiting, oral mucosal damage, altered taste. These problems would affect the enteral nutrition, further exacerbated malnutrition, reduced immunity and prolonged the hospital stay. The previous study showed that weight loss would shorten survival of cancer patients. Another study showed death usually occurs when there is 30% weight loss. The effectiveness of short‐term nutrition intervention in hospitalized cancer patients might be prolonged survival.

Objectives

The purpose of present study was to evaluate the effectiveness of nutritional intervention for hospitalized cancer patients. To evaluate the relationship between three‐year survival rate of cancer patients and the effectiveness of nutritional intervention by survival analysis.

Methods

This was a retrospectively observational study. The medical records of cancer patients receiving nutrition intervention during hospitalization were reviewed (n=291). Patients’ anthropometric measurements and blood biochemical parameters were recorded at the first, second and third nutritional intervention within a single hospitalization. The patients were further tracked three‐year survival (n = 214), and carried out the relevant factors of survival analysis.

Results

Total Patients’ energy intake and percentage of TER (Percentage of Total Energy Requirement) were significantly increased after nutritional intervention (both p = 0.000). There were no significant differences in BMI (Body Mass Index) and blood biochemical parameters related nutrition during hospitalization. The survival analysis in be tracked three‐year survival patients was performed. Those patients’ percentage of TER were significantly increased after nutritional intervention, it was the same with all patient. Patients’ percentage of TER at third nutrition intervention higher, patients’ survival rate better (p=0.026).

Conclusions

The cancer patients’ total energy intake could be improved by nutritional intervention; it might be helpful to survival.
L‐Cysteine Supplementation Increases Adiponectin Synthesis and Secretion, GLUT4 and Glucose Utilization by upregulating Disulfide Bond A‐Like Protein Mediated by MCP‐1 and TNF‐α Inhibition in 3T3‐L1 Adipocytes Exposed to High Glucose
Authors: Achari Arunkumar E , Jain Sushil K
Published: November 2023

Adiponectin is an anti‐diabetic and anti‐atherogenic adipokine; its plasma levels are decreased in obesity, insulin resistance, and type 2 diabetes. An adiponectin‐interacting protein named disulfide bond A‐like protein (DsbA‐L) play an important role in the assembly of adiponectin This study examined the hypothesis that L‐cysteine (LC) regulates glucose homeostasis through the DsbA‐L upregulation and synthesis and secretion of adiponectin in diabetes. 3T3L1 adipocytes were treated with LC (250 and 500 μM, 2 h) and high glucose (HG, 25 mM, 20 h). Results (n=3) showed that LC supplementation significantly (p < 0.05) upregulated the DsbA‐L, adiponectin and GLUT‐4 protein expression and glucose utilization in HG treated adipocytes. LC treatment along with insulin enhanced glucose utilization compared to either insulin or LC alone in HG treated cells (n=2). LC supplementation significantly (p < 0.05) promoted the secretion of total and HMW adiponectin secretion in HG‐treated adipocytes (n=5). In addition, LC significantly (p < 0.05) decreased ROS production and MCP‐1 and TNF‐α secretion in HG treated cells (n=3). We further investigated whether MCP‐1 and TNF‐α has any role of LC on DsbA‐L expression and adiponectin levels in 3T3‐L1 cells. Treatment with LC prevented the decrease in DsbAL (n=4) adiponectin (n=3) and GLUT‐4 (n=3) expression in 3T3L1 adipocyte cells exposed to MCP‐1 and TNF‐α. Thus, this study demonstrates that DsbA‐L and adiponectin upregulation mediates the beneficial effects of LC on glucose utilization by inhibiting MCP‐1 and TNF‐α secretion in adipocytes and provides a novel mechanism by which LC supplementation can improve insulin sensitivity in diabetes.

Support or Funding Information

The authors are supported by grants from the National Institutes of Health RO1 AT007442 and Malcolm Feist Endowed Chair in Diabetes
Impact of 15kDa Selenoprotein and Dietary Selenium on Initiation and Promotion in Colorectal Carcinogenesis
Authors: Tsuji Petra A , Canter Jessica A , Carlson Bradley A , Margulies Derek , Patterson Angelica , Gladyshev Vadim N , Yu Yunkai , Cao Liang , Davis Cindy D , Hatfield Dolph L
Published: November 2023

The micronutrient selenium (Se) largely exerts its biological effects through incorporation into selenoproteins. Several selenoproteins, including the 15kDa selenoprotein (Sep15), are thought to play roles in both cancer prevention and promotion. Previously, Sep15 knockout (KO) mice were shown to be resistant to formation of chemically‐induced pre‐neoplastic lesions. In our current study, Sep15 KO mice and littermate controls were maintained on deficient (0.02 ppm) or adequate (0.1 ppm) dietary Se. Mice were injected with azoxymethane (AOM) or saline, followed by treatment with dextran sulfate sodium (DSS) or water, to initiate and promote colon tumorigenesis. Formation of pre‐neoplastic lesions was again dramatically reduced in Sep15 KO mice compared to littermate controls, independent of dietary Se levels. Fewer Sep15KO mice developed tumors when Se‐deficient, but formed a similar number of tumors as controls when Se‐replete. Tumor mass itself appeared to be similar in control and Sep15KO mice when Se‐deficient, and slightly larger in Sep15 KO mice when Se‐replete. This suggests that Sep15 KO mice may be protected against AOM‐induced cancer initiation, but not against AOM/DSS‐induced tumor formation, and that dietary Se may modify the outcome. To assess cancer initiation, hepatic mRNA expression of CYP2E1, the enzyme responsible for activating AOM to its mutagenic form, was quantitated, and found to be significantly reduced in untreated Sep15 KO mice compared to littermate controls in Se‐deficient conditions, whereas no differences were detected in AOM/DSS‐treated mice. Interestingly, there also appear to be Se‐mediated differences in hepatic CYP2E1 expression. Inflammation is a strong tumor promoter in colorectal cancer, and pro‐inflammatory serum cytokines were generally higher in mice treated with AOM/DSS compared to those injected with saline. Interestingly, serum interferon‐γ, TNFα and Cxcl1 levels did not differ between Sep15 KO mice and controls, regardless of dietary Se or inflammation treatment. Other cytokines are currently being assessed in serum, colorectal epithelia, and spleens.

Support or Funding Information

NCI Intramural support; NIH CA080946 (VNG); Office of Dietary Supplements; Towson University Jess & Mildred Fisher Endowed Chair (PAT)
The AHR Pathway And Its Potential Role In The Regulation Of The 15 kDa Selenoprotein
Authors: Galinn Sarah E , Rosso Lauren E , Carlson Bradley A , Tobe Ryuta , Naranjo‐Suarez Salvador , Tsuji Petra A
Published: November 2023

Several selenoproteins, including the 15kDa selenoprotein (Sep15), are thought to have roles in both cancer prevention and promotion. Sep15 belongs to the group of stress‐related selenoproteins, and though its functional role continues to be elucidated, this oxidoreductase resides in the ER and is thought to be involved in reduction or isomerization of disulfide bonds. Little is known about factors regulating Sep15 expression. Our preliminary analyses indicated that some dietary polyphenols decreased Sep15 mRNA expression in human colon cancer HT29 cells. Because many of these flavonoids are known to affect Arylhydrocarbon Receptor (AHR) and/or NRF2‐regulatory pathways, expression of AHR and NRF2‐related genes are being investigated in Sep15‐deficient and ‐sufficient HT29 colon cancer cells to elucidate the potential regulatory mechanisms of Sep15. Analysis of previously published microarray data indicated that mRNA expression of AHR‐pathway‐related genes was significantly changed in HT29 cells lacking Sep15 compared to controls, including the AHR‐nuclear transporter, the AHR‐interacting protein, and heat‐shock protein 90. Comparison of the NCI‐60 cell lines through CellMiner analysis revealed that mRNA expression of Sep15 and AhR was negatively correlated (p<0.05). Furthermore, treatment of HT29 cells with the NRF2‐agonist, sulforaphane, did not increase Sep15 mRNA expression. Thus, in the current study, HT 29 control and shSep15 cells were exposed to 2,3,7,8‐Tetrachlorodibenzo‐p‐dioxin (TCDD) or 5,6‐beta‐naphthoflavone to induce or inhibit the AHR‐signaling pathway. Gene expression is currently being assessed using qPCR and Western blotting. Preliminary analyses show that mRNA expression of TCCD‐induced CYP1A1 and CYP1B1 is significantly lower in Sep15‐deficient cells compared to controls. Independent of TCDD treatment, shSep15 cells also expressed a lower CYP1A2 mRNA expression than the control cells. Nuclear and cytosolic protein expression is currently being assessed. Preliminary results suggest that Sep15 may influence or be regulated by AhR‐dependent pathways, which may be important in regulation of colon carcinogenesis.

Support or Funding Information

National Cancer Institute Intramural support; Towson University Jess & Mildred Fisher Endowed Chair and a Faculty Development & Research Committee grant (PAT).
The association of vitamin D level with dietary Se intake in elderly Korean: based on the 2008–2011 Korean National Health and Nutrition Examination Survey
Authors: Lee Okhee , Yang YoonKyoung , Shin Juyoung
Published: November 2023

The essentiality of vitamin D and Se in human nutrition have been well known. Their deficiencies in elderly were commonly suggested to increase the risks of cardiovascular disease, osteoporosis, dementia, sarcopenias etc. The dietary Se intake depends on the Se content of consumed foods and dietary pattern of a population. The food Se contents depend on Se status in soil, so they show wide variation even in the same food item. Due to its volatile nature, Se is difficult to analyze and needs specific caution when instrumentally chemical digestive analysis method is used. Therefore, the suitable database of Se for Korean foods is rarely observed. In this study, we analyzed the association of blood 25(OH) vitamin D level of 2008–2011 KNHANES datasets with dietary Se intake calculated using the food Se contents analyzed by INAA method. INAA is non‐destructive method which is no digestive method. The elderly with age over 60 year and participated in all the measurements of DXA, nutrition survey, and blood analysis were chosen. The dietary intake measured by 24‐hour recall method from nutrition dataset were used to calculate the selenium intakes of elderly. The food Se contents were coded using the KNHANES food database and used to calculate the dietary intake of elderly participated in KNHANES survey. The statistical analysis was conducted using SAS program (ver. 9.4) and considering the complex clustered sampling. The age‐standardized prevalence of Se intake deficiency in elderly Koreans over age 60y have a range from 39% to 63%, depending on age and gender. The lowest Se ingestions were found in both gender of elderly who have the age‐adjusted and unadjusted 25(OH) vitamin D level under 10ng/ml (p<0.001). In conclusion, this study suggested that blood 25(OH) vitamin D level is associated with dietary Se intake in aged Korean.

Support or Funding Information

Acknowledgement: This work was supported by the National Research Foundation through Basic Study Support Program 2014(NRF‐2014‐0023314, NRF‐2013‐0023314, NRF‐2012‐0023314, NRF‐2010‐0023314) and Nuclear Research and Development Program (NRF‐2009‐0078272).

The prevalence of deficient Se intake in elderly Korean

Age group
β
SE(β)
t Value
Pr > |t|

60=<age<70
0.393
0.012
31.8
<.0001

70=<age<80
0.573
0.016
36.81
<.0001

80=<age
0.628
0.034
18.57
<.0001
Associations of pregnancy folate and choline status with parental cancer risk factors and offspring's stem cell markers
Authors: Shen Yu‐Li , Huang Kevinta , Lee Chien‐Nan , Cheng Chin‐Pao , Chen Ling‐Ru , Ku Wei‐Chi , Huang Rwei‐Fen S.
Published: November 2023

Aims of the study were to investigate the associations of pregnant folate and choline status with parental diseases risks and stem cell markers which may involve in metabolic programming for offspring's susceptibility to cancer diseases. Fifty pregnant women were recruited from multiple clinic centers and NTU hospital to participate through the study. Blood samples from the first trimester pregnant women were collected for measurements of serum folate, choline, betaine and B12 levels. Lymphocytic DNAs were extracted for methylenetetrahydrofolate reductase (MTHFR) genotyping and for methylated promoter status of sonic hedgehog (SHH), insulin‐like growth factor‐2 (IGF2) and LINE‐1 (long interspersed nucleotide elements). Dietary data were collected by qFFQ and analyzed for nutrient intakes. The data revealed that deficiency rates of total choline (< 50% AI) and folate intake (< EAR) among the pregnant women were 11% and 14%, respectively. MTHFR 677CT/TT genotypes in relative to CC wild type were associated with higher intakes of phosphotidylcholine and total choline, and with lower serum folate (P = 0.049) and betaine (P = 0.023) levels. Historical parental obesity in relative to normal‐weight parents was associated with higher intake levels of folate (P = 0.002) and total choline (P = 0.048), and with significantly lower plasma free choline levels (P = 0.049) in pregnant off springs. Serum choline and folate levels of pregnant offspring were differentially associated with lymphocytic methylation status of SHH, IGF2 and LINE‐1 and their parental cancer risk factors.
Characterization of Cystathionine Beta‐Synthase and Cystathionine Gamma‐Lyase in the Production of Hydrogen Sulfide Biomarkers, Lanthionine and Homolanthionine, in a HepG2 Cell Culture Model
Authors: DeRatt Barbara , Ralat Maria , Gregory Jess
Published: November 2023

Hydrogen sulfide (H2S) is an essential gasotransmitter with the ability induce vasodilation and exert other cardioprotective effects in‐vivo, yet accurate quantification of H2S is difficult and often erroneous. Proposed biomarkers, lanthionine and homolanthionine, are produced concurrently with H2S by the pyridoxal 5′‐phosphate (PLP) dependent‐transsulfuration enzymes, cystathionine beta‐synthase (CBS) and cystathionine gamma‐lyase (CSE). This study sought to determine the contribution of CBS and CSE in the production of H2S biomarkers in a HepG2 cell culture model. Isotopic tracers, [U‐13C5] L‐methionine and (3, 3 D2) L‐cysteine, were used to determine metabolic fluxes through the transsulfuration pathway before and after the inhibition of CSE by 1mM propargylglycine (PAG), a CSE specific inhibitor. Cellular methionine concentrations and remethylation kinetics were not affected by PAG addition, nor was extracellular cysteine pool size. CSE inhibition caused intracellular cystathionine to accumulate (P=0.012). We evaluated the CBS‐catalyzed appearance of [D2]‐cystathionine and [13C4]‐cystathionine from homocysteine condensation with cysteine or serine, respectively. The labeling pattern of cellular cystathionine showed that only 4% of the cystathionine pool was derived from the CBS‐catalyzed condensation of homocysteine with cysteine. Lanthionine concentrations were four times greater in cells with PAG compared to control cells (P<0.001) but enrichments were not affected by PAG addition. This suggests CSE does not contribute to lanthionine synthesis but CSE may catalyze some lanthionine cleavage. Homolanthionine production in control cells greatly exceeded that observed in PAG‐treated cells (P<0.001), which confirmed that CSE is essential in homolanthionine synthesis. These results support our primary human hepatocyte studies which showed that CSE is the primary source of homolanthionine production while CBS is solely responsible for lanthionine production. These data illustrate how this HepG2 model is useful for mechanistic investigation of H2S production.

Support or Funding Information

Supported by NIH DK072398.
Comparison of Oral Iron Supplement Formulations for Normalization of Iron Status Following Roux‐en‐Y Bariatric Surgery
Authors: Mischler Renee , Armah Seth , Rosen Arthur , Choi Jennifer , Gletsu‐Miller Nana
Published: November 2023

Background

Iron deficiency is common after gastric bypass (GB) and treatment with oral supplementation using non‐heme iron, although currently recommended, is controversial. The study objective is to compare the effectiveness of oral iron supplementation using non‐heme versus heme iron for repletion of iron deficiency.

Methods

Using a randomized, controlled, single‐blind study design, women who had GB and were iron deficient received non‐heme iron (FeSO4, 65 mg t.i.d.) or heme iron (12 mg t.i.d.). Measures of iron status, including blood concentrations of ferritin, total iron binding capacity (TIBC), soluble transferrin receptor (sTfR), and hemoglobin were determined at baseline and at 2, 4, and 8 weeks after supplementation. Data is presented as mean ± SE, or mean (95% CI).

Results

The study is ongoing, but preliminary findings are presented herein. Baseline characteristics of screened patients (N=28) are age: 40.4±1.4 years; BMI: 34±1.4 kg/m2; time since surgery: 5.7±0.6 years; ferritin: 38.0 μg/L (16.1, 59.9); TIBC: 379 μg/dL (352, 405); sTfR: 2368.1 μg/L (1583.6, 3152.7); hemoglobin: 12.1 g/dL (11.5, 12.6). Of screened patients, 57.1% were classified as iron deficient and 35.7% had anemia. To date, 8 iron deficient patients have completed the 8‐week study (4 per group). Of those who received non‐heme iron supplementation, an improvement in blood concentrations of hemoglobin (p= 0.01), TIBC (p< 0.001), and sTfR (p= 0.049) was seen; ferritin tended to improve (p= 0.068). Of those who received heme iron supplementation, no change in iron status was seen in any of the iron status biomarkers.

Conclusion

Early data demonstrate that non‐heme iron supplementation appears to be effective for repletion of iron status. Findings may guide treatment of iron deficiency, which is very common following bariatric surgery.

Support or Funding Information

International Copper Association USDA, ARP Assistantship, to RAM
Vitamin B12 Status and Infant Outcome in a Community‐Based Folic Acid Intervention Study
Authors: Hausman Dorothy B , Holder Angela , Park Hea Jin , Rosa Gisselle , Andersen Victoria , Fleming Jennifer , Kauwell Gail , Caudill Marie , Malysheva Olga , Fisher Susan , Bailey Lynn B
Published: November 2023

Adequate vitamin B12 intake and stores during pregnancy are necessary to maintain maternal status and for adequate nerve myelination and hepatic stores in the developing fetus. Additionally, since the vitamin B12 and folate metabolic pathways are interrelated, a vitamin B12 deficiency can cause a functional folate deficiency that increases risk for neural tube defects during development. The objective of this study was to assess vitamin B12 status of women participating in a double blind randomized‐controlled folic acid supplementation study performed in collaboration with the Athens Regional Nurse‐Midwifery Practice, Athens, GA. Participants (n=49), recruited through this community‐based practice, were randomly assigned to take daily prenatal supplements containing 400 μg or 800 μg folic acid and all vitamins (including 6 mcg vitamin B12), minerals and DHA required for a healthy pregnancy beginning at the first prenatal visit (<12 wk gestation). Maternal blood samples collected at baseline, 28 weeks, and delivery, and cord blood samples at delivery were analyzed for plasma vitamin B12 and serum and RBC folate concentrations. At baseline, vitamin B12 concentrations < 200 pg/ml were detected in 3/49 women (6%). In those completing the intervention (n=28), vitamin B12 concentration decreased throughout the pregnancy and the concentration at baseline was highly correlated with those at 28 wk (r=0.739, p<0.001) and at delivery (r=0.0645, p<0.001). Vitamin B12 concentration in cord blood was higher than (p<0.0001) and highly correlated (r=0.755, p<0.0001) with maternal B12 concentrations at delivery. No differences were detected in maternal or cord blood vitamin B12 concentrations related to the dose of supplemental folic acid. In unadjusted analysis, gestational age at delivery was related to maternal vitamin B12 concentration (r=0.421, p<0.05) and baseline vitamin B12 correlated with serum folate concentration (r=0.542, p < 0.005). Vitamin B12 concentration in cord blood was not related to gestational age, infant height, weight, head circumference or Apgar score, or maternal or cord blood serum or RBC folate status. In this study, women retained their relative vitamin B12 status despite being supplemented with the same level of the vitamin, suggesting that optimal vitamin B12 status prior to pregnancy is essential for maintaining adequate vitamin B12 nutrition for the pregnant woman and her developing fetus. Accordingly, vitamin B12 interventions should be initiated pre‐conceptionally to optimize status during pregnancy particularly among those at greatest risk for low vitamin B12 status.

Support or Funding Information

Support provided by Georgia Experimental Agricultural Station, HATCH #GEO00706 and #GEO00707, Obesity Initiative at the University of Georgia (UGA) and UGA Office of Vice President for Research.
Glycation of transferrin affects chromium binding and transport
Authors: Vincent John B. , Deng Ge , Dyroff Samantha L.
Published: November 2023

Chromium, as the trivalent ion, can act as a pharmaceutical agent, increasing the sensitivity of the body to insulin and improving the symptoms of conditions, such as type 2 diabetes, that result from insulin‐insensitivity. For chromium to act, it must enter insulin‐sensitive tissues including the skeletal muscle where most glucose metabolism occurs. Chromium is transported from the bloodstream (where it is stored) to the tissues by the protein transferrin. However, under conditions of high blood sugar concentrations (as would be experienced by a diabetic subject), transferrin is chemically modified by the blood glucose. This modification alters the ability of transferrin to bind chromium and to take the shape required for the transferrin to be bound and uptaken by cells to deliver the chromium. The rate at which chromium binds to transferrin is also altered. This should lead to a decreased ability of the chemically modified (glycated) transferrin to deliver chromium to tissues. The transport of chromium by transferrin and glycated transferrin in rats has also been examined as well as the effects of insulin stimulation of transferrin movement on glycated transferrin compared to unglycated transferrin. The results suggest the dose of chromium anticipated to be beneficial in increasing insulin sensitivity in diabetic subjects could be larger than that for healthy subjects.

Support or Funding Information

This work was funded in part by a University of Alabama College of Arts and Sciences Undergraduate Creativity and Research Activity award to S.D.
In Vitro Antioxidant Activities of Black and Red Raspberry Fruits after Sequential Solvent Extraction
Authors: Basu Paramita , Maier Camelia
Published: November 2023

A sequential solvent extraction was employed to compare the in vitro antioxidant activities of black and red raspberry fruit extracts. Total phenolics, flavonoids, and proanthocyanidins were determined in hexane (H), ethyl acetate (E), chloroform (C), methanol (M), and water (W) extracts of black (BR) and red (RR) raspberry fruits. In vitro antioxidant activities of extracts were evaluated by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS), nitric oxide (NO), superoxide anion (O2−) scavenging, ferric reducing antioxidant potential (FRAP) and ferric reducing power assays. The highest phenolic contents were found in BRE and RRM extracts (168.6 ± 0.32 and 164.7 ± 0.31 mg GAE/g, respectively). The highest flavonoid content was determined in BRM, BRW, and RRM extracts (69.8 ± 2.4, 71.4 ± 0.2 and 70 ± 2.1 mg QE/g, respectively). BRM and RRH extracts possessed the highest proanthocyanidin content (601.1 ± 0.8 and 165.3 ± 0.1 mg GAE/g, respectively). BRM extract (100 μg/ml) showed the highest ferric reducing power (A700 = 0.94 ± 0.002). The highest FRAP activity was detected in BRM extract (A593 = 1216 ± 8.4 μM Fe2+ g−1). In general, all extracts exhibited increasing radical scavenging activities with increasing concentrations (20–100 μg/ml). The highest DPPH, ABTS, O2− and NO radical scavenging activities were displayed by 100μg/ml of BRM (A517 = 92.2 ± 0.13), BRW (A734 = 83.4 ± 0.21), RRW (A230 = 93.1 ± 0.6) and RRW (A540 = 61.4 ± 0.33) extracts, respectively. Pearson correlation was performed between IC50 values of DPPH, ABTS, O2− and NO scavenging activities and total phenolics, flavonoids, proanthocyanidins contents of fruit extracts. The IC values of all radical scavenging assays of black raspberry extracts rather than those of red raspberry extracts correlated with total phenolics. Higher correlations of IC50 values of DPPH, ABTS, O2− and NO scavenging activities of methanol and water extracts of both fruits with total phenolic, flavonoids and proanthocyanidins than those of the other solvent extractions were found. This indicates that methanol and water are better solvents for extraction of potent antioxidants from black and red raspberry fruits that could serve as sources of natural food antioxidants.
Flaxseed Lignan Metabolite, Enterolactone, Down‐regulated DNA Methyltransferase, Histone Deacetylase, and Methyl‐CpG‐binding Domain Protein Expression in Murine Adipocytes
Authors: Rhee Yeong
Published: November 2023

Nutrition influences gene expression via epigenetic control mechanisms such DNA methylation and histone modification by DNA methyltransferase (DNMT) and histone deacetylase (HDAC), respectively. Oxidative stress caused by reactive oxygen species (ROS) damages DNA or cells which can be regulated by antioxidants. Flaxseed is the richest source of secoisolariciresinol diglucoside, which is metabolized into enterolactone by bacteria in the gut. Enterolactone has antioxidant activities which lower ROS induced oxidative damages. Thus, this study was conducted to determine the effects of enterolactone on DNMTs, HDACs, and methyl‐CpG‐binding domain protein (MBD) expression in ROS treated murine adipocytes. Enterolactone (0, 10, 50, or 100 μM) or control antioxidant, such as superoxide dismutase (100 U/mL) or catalase (100 U/mL), treated adipocytes (1×107) were treated with hydroxyl or superoxide radical, and then total RNA was extracted and purified using an RNeasy Mini kit. DNMTs, HDACs, and MBD2 expression was measured by real‐time quantitative PCR. The fold change in DNMTs, HDACs, and MBD2 expression was calculated using 2−ΔΔCT method. The results showed that enterolactone down‐regulates DNMTs, HDACs, and MBD2 expression of hydroxyl radical treated adipocytes. However, no significant effects of enterolactone on DNMTs, HDACs, and MBD2 gene expression were seen in superoxide radical treated adipocytes. How DNMTs, HDACs, and MBD2 expression of hydroxyl radical treated adipocytes was down‐regulated by enterolactone is unknown and needs further study.

Support or Funding Information

The research was funded by grants from the Department of Health, Nutrition, and Exercise Sciences and College of Human Development and Education at North Dakota State University.
The effect of cranberry consumption on lipid and lipoprotein metabolism in human apolipoprotein A‐I transgenic mice fed a high fat and high cholesterol diet
Authors: Park Young‐Ki , Caceres Christian , O'Neill Edward N. , Bae Minkyung , Pham Tho X. , Lee Yoojin , Kim Bohkyung , Lee Ji‐Young
Published: November 2023

Cranberries contain high levels of health‐promoting anthocyanins, which have been implicated in their potential role in the prevention of cardiovascular disease. The objective of this study was to determine the effects of cranberries on lipid and lipoprotein metabolism in mice expressing human apolipoprotein A‐I transgene (hApoAITg), which have similar HDL metabolism to that of humans. Male hApoAITg mice were fed a modified AIN‐93M high fat/high cholesterol control diet (HF/HC; 15% fat, 0.25% cholesterol, w/w; n = 15) or a HF/HC experimental diet supplemented with 5% (w/w) dried, whole cranberry powder (n = 16) for 8 weeks. Subsequently, the mice were sacrificed to obtain tissue and blood samples. There were no significant differences in terminal body weights, body weight gain, liver weights and epididymal fat weights between control and cranberry‐fed mice. Serum total cholesterol (TC) and apolipoprotein B cholesterol (ApoB‐C) levels were significantly lower in controls than cranberry group (238 ± 9.4 vs. 294 ± 20.3 mg/dL for TC; 43.9 ± 6.7 vs. 88.3 ± 15.0 mg/dL for ApoB‐C) with no significant difference in serum high‐density lipoprotein cholesterol (HDL‐C) concentrations. Serum triglyceride (TG) levels of cranberry group (55.5 ± 4.6 mg/dL) were also significantly higher than controls (37.7 ± 2.9 mg/dL). Hepatic expression of sterol‐regulatory element binding protein 2 and low‐density lipoprotein receptor was significantly lower in cranberry group, which may be responsible for the increased serum TC and apoB‐C, while HMG‐CoA reductase and proprotein convertase subtilisin/kexin type 9 mRNA levels were not different between two groups. Despite the significant increase in serum TG in mice‐fed cranberries, their mRNA abundance of genes responsible for lipogenesis (e.g., fatty acid synthase and stearoyl CoA desaturase) and fatty acid oxidation (e.g., carnitine palmitoyl transferase 1α and acyl‐CoA oxidase 1) was not significantly altered in the liver. In addition, the hepatic expression of genes involved in HDL metabolism, including human apoA‐I, lecithin:cholesterol acyltransferase, and scavenger receptor class B, type I, was significantly lower in cranberry‐fed mice compared with controls although serum HDL‐C levels were not different between two groups. Further study should be warranted to have better mechanistic insight into the changes in lipid and lipoprotein metabolism by cranberries.

Support or Funding Information

Nutricia Research Foundation
Lipotoxicity‐Evoked Reductions in Nitric Oxide Bioavailability and Exaggerated Oxidant Stress Are Blunted By Blueberry Metabolites in Human Vascular Endothelial Cells
Authors: Bharat Divya , Begaye Nathan , Ramos Renata , Ferreira Marina , Li Youyou , Cutler Brett , Bharath Leena P , Symons J David , Jalili Thunder , Pon Velayutham Anandh Babu
Published: November 2023

The vascular benefits of blueberry (BB) anthocyanins reported in humans might be mediated by their circulating metabolites. To test this, we synthesized the BB‐metabolites vanillic acid‐4‐sulfate (V4S), isovanillic acid‐3‐sulfate (IV3S), and benzoic acid‐4‐sulfate (B4S), because these compounds are not commercially available. As hypothesized, a combination of V4S, IV3S, B4S, hydroxyhippuric acid (HHA), and hippuric acid (HA) at concentrations known to peak in human plasma following the consumption of 240 g of blueberries, suppressed chemokines (IL8 and MCP1) and adhesion molecules (ICAM1, VCAM1 and E‐selectin) unleashed by human aortic endothelial cells (HAECs) upon treatment with palmitate (500 μM for 5 h) vs. vehicle. Next, we determined whether vasculoprotection afforded by BB‐metabolites was secondary to preserved endothelial cell nitric oxide (NO) bioavailability. HAECs were treated ± a BB‐metabolite cocktail for 6 h ± 500 μM palmitate for the last 5 h. To determine agonist‐stimulated NO production, HAECs were treated with 100 nM insulin (or vehicle) for 30 min at the end of the experimental treatment. Human studies reported that blueberry intake increases the endothelium dependent vasodilation that was accompanied by the increases in the plasma concentrations of metabolites such as V4S, IV3S, B4S, HA, and HHA. Hence, BB‐metabolite cocktail contained these metabolites at physiological relevant concentrations. Palmitate increased reactive oxygen species (ROS) generation (assessed by H2DCFDA) and blunted insulin‐stimulated NO production (assessed by DAF‐FM) in HAECs (both p<0.05), but both responses were normalized in BB‐metabolite‐treated HAECs. This is consistent with our results on the beneficial effects of BB‐metabolites on endothelial inflammation. Ongoing studies are determining the extent to which these findings from HAECs can be translated to intact arteries. In this regard, preliminary results from murine femoral arteries indicate that palmitate‐induced endothelial dysfunction can be attenuated by concurrent treatment with BB‐metabolites. Blueberry supplement might provide a valuable adjunct therapy to traditional treatments for vascular complications known to exist in patients with metabolic syndrome.

Support or Funding Information

University of Utah Research Start‐up fund (to Anandh Babu Pon Velayutham), Native American Summer Research Internship (to Nathan Bagaye), Undergraduate Research Opportunities Program assistantship (to Brett Cutler), and Brazilian Scientific Mobility Program sponsorship (to Marina Ferreira and Renata Ramos).
Characterization of the Ingredients in Fish Oils and Herbal Dietary Supplements analyzed by GC‐MS and HPLC‐ELSD
Authors: Ikediobi Stephanie , Nikolau Basil J.
Published: November 2023

The Food and Drug Administration (FDA) regulates dietary supplements differently than conventional drugs, which raises questions for consumers about dietary supplement integrity. Previous analyses by DNA fingerprinting indicated that dietary supplements might contain unreported contaminants (NYAG 2013 [Press Release]1). However there are limitations to DNA fingerprinting technology, in that intact DNA segments from 500–1000 bps need to be in the sample to accurately analyze the DNA fingerprint of the sample. Because many dietary supplements use production practices that heavily process the product, DNA fragments may not retain integrity, resulting in misidentification (Reynaud et. al. 20152). Our goal is to build on previously established biochemical analytical platforms and analyze dietary supplements using chromatographic methods such as Gas Chromatography‐Mass Spectrophotometry (GC‐MS) and/or High Performance Liquid Chromatography – Evaporative Light Scattering Detector (HPLC–ELSD) to quantitatively characterize the presence of primary “putative bioactive” ingredients, and compare these data to manufacturers’ claims. For example, oil supplements such as fish and krill oil are a rich source of omega‐3 fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and are typically consumed for cardiovascular health. We evaluated commercially available fish and krill oils by GC‐MS analysis of the fatty acid methyl esters (FAMEs), and the lipid profiles of these oils by HPLC‐ELSD analysis. These analyses indicate that some manufacturers under‐report or over‐report the bioactive ingredient amounts per capsule. These findings highlight the complexity by which the integrity of dietary supplements can be measured.
Improving Wound Healing through the Use of Tetrazine‐Modified Alginate
Authors: De Leon‐Peralta Esmarline Jacely , Brudno Yevgeny , Kwee Brian J. , Mooney David J.
Published: November 2023

Ischemic diseases, such as myocardial infarction and peripheral arterial disease, are a leading cause of death globally. There has been increasing interest in treating these diseases with factors that induce angiogenesis, the growth of new blood vessels from pre‐existing blood vessels. We have recently shown that alginate linked to tetrazine helps induce a greater recovery in blood perfusion compared to unmodified alginate in a murine model of hindlimb ischemia. The objective of this project is to understand how this biomaterial induces recovery in blood perfusion. Our hypothesis is that tetrazine acts directly on endothelial cells to induce angiogenesis and also activates immune cells that are proangiogenic. To validate our hypothesis, we have performed in vitro angiogenesis assays to study if tetrazine‐modified alginate induces endothelial cell sprouting. Additionally, we have performed histology and immunohistochemistry on ischemic muscles from our in vivo experiments to see if this biomaterial increases blood vessel density, blood vessel remodeling, and the presence of proangiogenic immune cells. Our findings may lead to a novel method for treating people with ischemic diseases.

Support or Funding Information

Wyss Institute for Biologically Inspired Engineering, Harvard John A. Paulson School of Engineering and Applied Sciences Research Experiences for Undergraduates (SEAS REU)
PERK Branch of the ER Stress Regulates FoxO to Mediate Homocysteine‐induced BKCa Channel Dysfunction
Authors: Sun Wen‐Tao , Wang Xiang‐Chong , Mak Shiu‐Kwong , Yu Cheuk‐Man , Yang Qin
Published: November 2023

Large‐conductance Ca2+‐activated K+ (BKCa) channels play an important role in vascular tone regulation. The atherosclerotic risk factor homocysteine inhibits vascular smooth muscle BKCa channels whereas the mechanisms remain poorly understood. We studied the role of endoplasmic reticulum (ER) stress in homocysteine‐induced BKCa channel inhibition with further understanding of the molecular determinants. Vasorelaxant response to the BKCa channel opener NS1619 was studied in endothelium‐denuded porcine small coronary arteries in a myograph. Primary cultured porcine coronary arterial smooth muscle cells (PCASMCs) were used for mRNA and protein analysis of BKCa channels, as well as patch‐clamp recording of the BKCa channel current. NS1619‐induced vasorelaxation was significantly inhibited by homocysteine, which was restored by ER stress inhibitors either TUDCA or 4‐PBA. GSK2606414, a selective inhibitor of PKR‐like endoplasmic reticulum kinase (PERK) showed potent protective effect against homocysteine on the NS1619‐induced relaxation and the whole‐cell BKCa channel current. Homocysteine lowered the protein level of β1 but not α subunit of the BKCa channel. Exposure to homocysteine did not alter the mRNA expression of both subunits. Downregulation of BKCa β1 was attenuated by TUDCA, 4‐PBA, and GSK2606414. Pharmacological inhibition of PERK suppressed the nucleic translocation of forkhead box O transcription factor 3a (FoxO3a) in homocysteine‐exposed PCASMCs, associated with a decrease of atrogin‐1 protein and an increase of BKCa β1 protein. In PCASMCs transfected with FoxO3a siRNA, downregulation of atrogin‐1 protein expression and restoration of BKCa β1 protein level was observed after homocysteine exposure. This study suggests that ER stress mediates homocysteine‐induced BKCa channel inhibition in coronary arteries and activation of FoxO3a by PERK underlies ER stress‐mediated BKCa channel inhibition through a mechanism involving ubiquitin ligase‐enhanced degradation of the channel β1 subunit.

Support or Funding Information

Supported by RGC/GRF CUHK4774/12M & CUHK14118414; Lui Che Woo Institute of Innovative Medicine ‐ CARE theme 8303303, and CUHK Direct Grant 4054182.
Enhanced generation of platelet‐activating factor‐receptor ligands via ethanol synergizes ultraviolet B mediated acute and systemic effects
Authors: Sahu Ravi P , Weyerbacher Jonathan , Ocana Jesus , Konger Raymond , Travers Jeffrey
Published: November 2023

Alcohol (ethanol) intoxication remains one of the most abused substance in the United States and other part of the world. In addition, an association between alcoholism and trauma has long been recognized. Moreover, thermal injury after alcohol exposure have been shown to exhibit higher morbidity and mortality in humans. However, the mechanism of alcohol‐related toxicity with or without thermal injury is not entirely clear. Several studies including ours have shown that exposure to pro‐oxidative stressors including ultraviolet B (UVB) generate the potent lipid mediator with platelet‐activating factor‐receptor (PAF‐R) ligand activity. Notably, these PAF‐R ligands mediate UVB‐induced acute pro‐inflammatory effects and also augment photosensitivity associated with the deficiency of xeroderma pigmentosum complementation group A. Given that several agents can modulate PAF‐R ligands production and systemic ethanol exposure enhances UVB‐induced effects, we hypothesize that ethanol can synergistically potentiate UVB‐mediated PAF‐R ligands production as well as acute pro‐inflammatory and systemic effects. Using the human keratinocyte‐derived cell line, HaCaT, we demonstrate that preincubation with ethanol resulted in an augmentation of PAF‐R ligands production in response to low (sub‐erythematic) doses of UVB. Pre‐treatment of C57BL/6 mice with systemic ethanol resulted in an augmented skin production of PAF‐R ligands in response to UVB. We demonstrate that pre‐treatment with systemic ethanol followed by UVB irradiation of PAF‐R positive wild‐type versus PAF‐R‐deficient mice resulted in enhanced skin erythema response compared to UVB alone treated group in a PAF‐R dependent fashion. Moreover, the combination of ethanol + UVB resulted in increased systemic toxicity, including lung inflammation only in wild‐type mice and not in the PAF‐R‐deficient mice. These studies demonstrate the involvement of the PAF‐R pathway in ethanol‐induced acute toxic effects and that ethanol can synergistically potentiate PAF‐R‐mediated systemic effects when combined with agents (such as UVB) exhibiting pro‐oxidative stressors property. These studies indicate that blocking PAF‐R‐mediated pathway could be used as a promising approach to prevent ethanol ± UVB‐mediated acute and systemic toxicities.

Support or Funding Information

NIH grant R01 HL062996 and K22 ES023850