bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti‐Diabetic Efficacy of a Novel Fenugreek Seed (Trigonella foenum‐graecum, Fenfuro) Extract in Patients with Type‐2 Diabetes
Authors: BAGCHI DEBASIS , Swaroop Anand , Preuss Harry G , Bagchi Manashi , Kumar Pawan
Published: November 2023

Fenugreek seeds are known to exhibit potent antioxidant, hypoglycemic and nephroprotective activities. Previous studies in our laboratories demonstrated the broad spectrum safety and efficacy of Fenfuro, a novel Trigonella foenum‐graecum seed extract enriched in furostanolic saponins, in type 2 diabetes (T2D) in rats. We conducted a multi‐center, randomized, placebo‐controlled, double‐blind, add‐on clinical study evaluated over a period of 3 months the efficacy of Fenfuro (500 mg bid) in 154 male and female subjects (25–60 years) with T2D. This study examined the body weight, blood pressure and pulse rate, as well as the efficacy of Fenfuro on fasting and post‐prandial plasma sugar, HbA1c and fasting and post‐prandial C‐peptide levels. Fenfuro caused significant reduction in both fasting plasma and post‐prandial blood sugar levels. Approximately 83% of the subjects reported decrease in fasting plasma sugar levels in the Fenfuro group as compared to 62% in the placebo group, while 89% of the subjects demonstrated reduction in post‐prandial plasma sugar levels in the Fenfuro‐treated group as compared to 72% in the placebo group. HbA1c levels were reduced in both placebo and treatment groups. The decrease in HbA1c levels was significant in both groups as compared to respective baseline values. A significant increase in fasting and post‐prandial C‐peptide levels compared to the respective baseline values, were observed, while no significant changes in fasting and post‐prandial C‐peptide levels were observed between the two groups. Extensive blood chemistry analyses exhibited the broad spectrum safety of Fenfuro. Furthermore, 48.8% subjects reported reduced dosage of anti‐diabetic therapy in Fenfuro‐treated group, while 18.05% reported reduced dosage of anti‐diabetic therapy in the placebo‐group. Overall, Fenfuro proved safe and efficacious in ameliorating the symptoms of T2D in humans.

Support or Funding Information

Study Supported by Cepham Research Center, Piscataway, NJ, USA
Development of Novel Hollow Zein Nanoparticles for Delivery of Nutraceuticals
Authors: Hu Siqi , Fernandez Maria‐Luz , Luo Yangchao
Published: November 2023

Zein is the major storage protein from corn with strong hydrophobicity and unique solubility and has been studied as an excellent natural biomaterial for encapsulation and delivery of nutraceuticals. Although zein nanoparticles can be easily prepared by anti‐solvent liquid‐liquid dispersion method, the nanoparticles usually have a dimension greater than 150 nm with a solid core, which greatly limited their loading capacity and cellular uptake for encapsulation applications. In this study, sodium carbonate was proposed as a sacrifice template and tannic acid was used as a natural emulsifier/stabilizer to prepare hollow zein nanoparticles (HZN). The formulation and fabrication process, including concentrations of zein and sodium carbonate, as well as mass ratio of zein to water, were optimized by surface response methodology (Box‐Behnken design). The optimal HZN was then comprehensively characterized and compared with solid zein nanoparticles (SZN). Our results indicated that both zein concentration and sodium carbonate concentration significantly affected the polydispersity index (PDI) and particle size, and the latter one was also significantly affected by zein/water ratio; while zeta potential was significantly affected by all three factors. The optimal HZN exhibited a small dimension of 82.5 nm with a PDI of only 0.103 and a zeta potential (surface charge) of −42.4 mV, indicating the nanoparticles were very homogenously distributed with excellent colloidal stability. Then, the lipophilic bioactive compound, curcumin, was adopted as a model nutrient to explore the encapsulation and delivery potentials of HZN, in comparison with SZN prepared under the same conditions. The molecular interaction and binding constant between curcumin and zein nanoparticles were investigated by Fourier transform infrared spectroscopy and fluorescent spectrophotometer. It was notable that the hydrophobic interactions and hydrogen bonding between curcumin and HZN was significantly stronger than that between curcumin and SZN. Meanwhile, the binding constant of curcumin to HZN was also greater that of SZN. Coincidently, when loaded at the same ratio (100:1, zein:curcumin), the encapsulation efficiency (98.6%) of HZN was dramatically higher than that of SZN (69.7%). The physical characteristics of curcumin‐encapsulated HZN almost remained the same having a size of 84.1 nm and PDI of 0.092, while the SZN showed a significantly larger particle size (157.4 nm) and PDI (0.239). The controlled release was tested in the simulated gastrointestinal fluids to mimic in vivo digestion. HZN showed a better retained release profile of curcumin than SZN. In summary, compared with traditional solid zein nanoparticles, the hollow zein nanoparticles developed in this study have promising features for oral delivery of lipophilic nutraceuticals.
The Impact of Online Dietary Assessment to Efficiency of Nutrition Counseling: A Pilot
Authors: Watowicz Rosanna P , Wexler Randell Keith , Weiss Rick , Spees Colleen Kelleher , Taylor Christopher Alan
Published: November 2023

Introduction

Nutrition counseling is an important intervention for many chronic diseases and dietary assessment is a crucial component of any initial nutrition counseling visit. Although necessary, the process of collecting a detailed dietary assessment is time consuming and, in turn, decreases the amount of time available for nutrition education and goal setting. The objective of this study was to examine whether an online dietary assessment completed prior to a nutrition counseling visit was related to more efficient nutrition counseling visits.

Methods

Twenty‐six adults were recruited out of primary care centers and were randomized to either the pre‐assessment group (n=13) or the control group (n=13). Participants in both groups then attended 2 nutrition counseling visits with registered dietitians in a local grocery store. The pre‐assessment group completed an online food frequency questionnaire prior to their first visit with the dietitian, to which the dietitian had access prior to the initial visit. The control group did not complete a dietary assessment before their visit. Following each visit, the dietitians reported the time spent on assessment, education, goal setting, and aisle tours, as well as the specific topics covered during the education portion. A depth score was also used to estimate the average depth of each of the topic covered in the session (1=basic introduction, 4=skill mastery activities).

Results

Participants who completed the online dietary assessment questionnaire experienced a more in‐depth nutrition educational experience as a result of the detailed preliminary report. When compared to the control group, the provision of the dietary intake assessment questionnaire resulted in significantly less time spent on assessment (14 vs. 19 minutes) and significantly more time spent on education (33 vs. 26 minutes) and goal setting (13 vs. 11 minutes) during the initial 1 hour session. Although the dietitians reported a shorter time spent on dietary assessment during the visit, they also reported spending an average of 13 minutes reviewing the online assessments prior to the visit. Those who completed the assessment prior to their visits had a greater mean number of topics covered, as well as a greater average depth score.

Discussion

Completing a dietary assessment prior to the initial nutrition counseling visit allowed for a more efficient visit with the dietitian. More time was spent on education and goal setting and the breadth and depth of the initial visit was greater for those in the pre‐assessment group. Requesting that patients complete a detailed dietary assessment prior to their nutrition counseling visits may be a feasible method for improving nutrition care, however additional research testing this model on a wider scale is necessary.

Support or Funding Information

The Ohio State University Food Innovation Center
Effects of quinoa (Chenopodium quinoa) consumption on fecal microbiome of obese diabetic (db/db) mice
Authors: Noratto Giuliana , Garcia‐Mazcorro Jose , Mills David
Published: November 2023

Quinoa, a gluten‐free pseudocereal native from the Andes, is known for its high nutritional value. Our aim was to investigate the effect of quinoa consumption on fecal microbial ecology of db/db mice.

The db/db mice were fed AIN‐93G diet (obese) or AIN‐93 supplemented with quinoa (quinoa) for eight weeks. Lean counterpart controls were fed with AIN‐93G diet (lean). Fecal bacterial communities were analyzed using gene sequencing of 16S rRNA. Data was analyzed with Quantitative Insights into Microbial Ecology (QIIME) v.1.8.0. Predicted metabolic profiles were analyzed with the online Galaxy version of PICRUSt. Short chain fatty acids (SCFAs) in cecal contents were analyzed with high performance liquid chromatography.

Results showed that quinoa consumption induced significant differences in bacterial abundances. Quinoa group showed lean‐like abundances in Enterococcus, Turicibacter, and Akkermansia. The later has been negatively correlated with body weight. Interestedly, quinoa consumption delayed body weight gain during the first five weeks of dietary supplementation. Analysis of weighted UniFrac distances revealed a distinctive clustering of lean microbial communities from obese and quinoa groups (ANOSIM, P < 0.01). Contrasting with these results; the weighed analysis showed that quinoa group clustered separately from obese and lean. Out of the predicted metabolic functions, most significant differences were found in lean compared to obese and quinoa groups. Quinoa consumption was associated with lower cecal butyrate and succinic acid concentrations.

These findings strongly indicate the complex interactions between diet, genetics, and gut microbial ecology, however further studies are needed to explore the clinical relevance of such interactions.

Support or Funding Information

New Faculty Seed Grant (# 3057‐9907)
Bioavailability of Long Chain Omega‐3 Polyunsaturated Fatty Acids from Phospholipid‐Rich Herring Roe Oil in Men and Women with Mildly Elevated Triacylglycerols
Authors: Cook Chad M , Hallaråker Hogne , Sæbø Per Christian , Innis Sheila M , Kelley Kathleen M , Sanoshy Kristen D , Berger Alvin , Maki Kevin C
Published: November 2023

Background

The bioavailability of long chain omega‐3 polyunsaturated fatty acids (LC n‐3 PUFA) may differ depending on the source and processing of the oil.

Objective

This randomized, double‐blind, crossover trial assessed the bioavailability of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA) from two sources.

Methods

Thirty‐two adults with fasting triacylglycerol (TAG) concentrations between 100–399 mg/dL were randomly assigned, with stratification by sex and age, to receive 12 capsules/day containing either phospholipid (PL)‐rich herring roe oil (Romega® 30, 628 mg/day EPA; 1810 mg/day DHA; 137 mg/day DPA) or TAG‐rich fish oil (575 mg/day EPA; 1843 mg/day DHA; 259 mg/day DPA) each for a 2‐week period separated by a 4 week washout. EPA, DHA, and DPA were measured in plasma phosphatidylcholine (PC) over a 12 h period following consumption of a single serving of each product, and in fasting blood samples obtained at the end of each 2‐week supplementation period.

Results

Thirty‐one subjects completed the study and were included in an efficacy evaluable analysis. A majority were non‐Hispanic white (71.0%) men (n = 15) and women (n = 16) with a mean ± SEM age of 45.8 ± 2.0 y, BMI of 27.1 ± 0.4 kg/m2, and fasting TAG of 167 ± 9.8 mg/dL. The net incremental (ni)AUC0–12 h for EPA (161 vs. 97 μg*h/mL), DHA (87 vs. 40 μg*h/mL), and EPA+DHA (239 vs. 126 μg*h/mL) in plasma PC were significantly higher (p < 0.01 for all) after Romega 30 supplementation compared to fish oil. Similar results were observed when the niAUC0–12 h for plasma PC EPA, DHA, and DPA in response to the Romega 30 condition were mathematically normalized to account for differences in the intake of these fatty acids in the TAG‐rich fish oil (p < 0.001 for all). There was a significant increase (change from baseline) in fasting EPA (36 ± 3 μg/mL Romega 30; 31 ± 4 μg/mL fish oil), DHA (88 ± 7 μg/mL Romega 30; 80 ± 9 μg/mL fish oil), and DPA in plasma PC (p < 0.0001 for all) after 2 weeks of daily supplementation within both conditions, but there were no statistically significant differences between conditions. Both products were well tolerated, as there were no adverse events related to study product consumption, no differences in vital signs or body weight measurements between the two conditions, and the frequency of gastrointestinal symptoms reported before the study were not different from similar measures taken at the end of each test period for each condition.

Conclusion

These data demonstrate that PL‐rich immature herring roe is a well‐tolerated and highly bioavailable source of EPA, DHA, and DPA LC n‐3 PUFA.

Support or Funding Information

Supported by Arctic Nutrition AS (Ørsta, Norway)
Long‐Term Vitamin D Deficiency Decreases Bodyweight and Adipose Tissue Mass in Female Mice
Authors: Belenchia Anthony M , Rosenfeld Cheryl , Johnson Sarah Ashley , Kieschnick Alyssa , Peterson Catherine
Published: November 2023

There is a well‐established inverse association between vitamin D status and adiposity. Moreover, the involvement of vitamin D and its receptor in the development of adipose tissue is undeniable. However, there is a paucity of direct in vivo data observing the consequences of prolonged vitamin D deficiency on body composition. In order to determine how long‐term vitamin D deficiency affects weight gain and adiposity, 8‐week‐old female C57bl/6 mice were randomized to either a control (AIN‐93G; CD) or vitamin D‐deficient (modified AIN‐93G; VDD) diet (t=0). Bodyweight was measured weekly beginning at randomization (t=0). At 40 weeks (t=40) body composition was determined by EchoMRI. Following euthanization, perigonadal and subcutaneous fat pads were excised and weighed. There were no between group differences in bodyweight at any time point until t=29, after which point VDD mice weighed significantly less than controls in each subsequent measurement. VDD mice had less total adipose tissue (p=0.031) as well as smaller perigonadal and subcutaneous fat pads than controls (p=0.014 & p=0.022, respectively). However, there was no difference in body fat percentage between the two groups (p=0.067) nor were there differences in the relative weights of either subcutaneous or perigonadal fat pads (p=0.119 & p=0.147, respectively). These data suggest that vitamin D is involved in adipose tissue maintenance; while the lack of differences in percent body fat or relative fat pad weights indicate that the observed weight loss in VDD mice is not attributable to fat loss alone.
Effects of apple snacking on body weight status and metabolic profiles of overweight and obese children
Authors: Cabalbag Conralyn , Metti Dina , Tunstall Ashley , Almaiman Lama , Hong Mee Young , Kern Mark , Hooshmand Shirin
Published: November 2023

Childhood obesity prevalence has increased and has been associated with continued obesity into adulthood with adverse health consequences. Consumption of fruits and dietary fibers have been shown to produce favorable impacts on risk for future chronic diseases such as cardiovascular disease (CVD) incidence. The objective of this study was to determine the potential for apple as a high‐fiber, nutrient‐rich snack to reduce body weight and improve associated comorbidities in overweight and obese children. Thirty‐three overweight or obese children ages 10 to 16 years with BMI ≥85th percentile of the CDC growth chart were randomly assigned to one of two groups. Individuals were provided with twice daily 120 kcal serving snacks of dried apple or a refined carbohydrate‐rich snack (muffins) for eight weeks. Body weight and waist circumference were determined at an initial visit and after the eighth week. Blood samples were collected at baseline and the final visit to measure serum cholesterol, triglycerides, glucose, insulin, proinsulin, total adiponectin, and C‐reactive protein. Both apple and muffin groups did not pose significantly different results in terms of body weight; however, waist circumference tended (P=0.07) to increase within the muffin group. Biochemical markers were not significantly different between groups; however, total adiponectin concentration increased (p<0.05) after the eight‐week treatment within the apple group. Overall, apple snacks did not promote adiposity; whereas, a refined snack tended to increase waist circumference. Furthermore, apple consumption increased concentrations of adiponectin, which is associated with lower bodyweight and reduced CVD risk. These results provide promising results for future research of other populations or for a longer duration.

Support or Funding Information

U.S. Apple Association
Anti‐tumor Properties of Beta‐sitosterol in AGS Human Gastric Adenocarcinoma cells and Tumor Xenograft Mice
Authors: Shin Eunju , Kim Sung Hee , Hwang Jin‐Taek
Published: November 2023

Although beta‐sitosterol (BS), one of several phytosterols, has been proposed to be effective for improving human health, the precise molecular mechanisms of BS regarding anti‐cancer effects have not yet been elucidated. Here, we investigated effects of BS regarding apoptotic mechanisms. BS significantly induced cell death in a dose dependent manner. These effects were accompanied by typical appearances of apoptosis including DNA fragmentation, apoptotic assay, caspase3/7 activity, mitochondria membrane potential and activation of tumor suppress genes including PTEN and AMPK in AGS cells. Inhibition of AMPK by compound C (AMPK inhibitor) rescued PTEN expression induced by BS. These results suggest that stimulation of PTEN expressions by BS is dependent on AMPK signaling. Furthermore, BS at 100mg/(kg · day) also inhibited tumor growth in tumor xenografts. Taken together, BS exerts potent anti‐cancer properties by modulating AMPK and PTEN in AGS cells as well as xenograft mice models.
Delta Tocotrienal Inhibit mTOR Pathway by Modulating Glutamine Uptake and Transporters in Non‐Small Cell Lung Cancer
Authors: Rajasinghe Lichchavi Dhananjaya , Gupta Smiti V.
Published: November 2023

Lung cancer, with the majority of cases being non‐small cell lung cancer (NSCLC), is one of the leading causes of death among cancers. NSCLC has exhibited high rate of glutamine dependency during its growth and development. It has also been shown that glutamine uptake plays a mandatory role in the uptake of essential amino acid and in activation of mTOR kinase. SLC1A5 and SLC7A5, glutamine transporters, exhibit an important role in the development of NSCLC by transporting glutamine and essential amino acid into the proliferating tumors. Therefore, SLC1A5 and SLC7A5 are key drug targets with potential pharmacological importance. The aim of this study was to probe dysregulated glutamine and essential amino acids (EAA) metabolism in NSCLC, while investigating the effect of T3 on glutamine transporter and mTOR pathways. H1299 and A549 cells were cultured with/without T3 (30μM) followed by extraction of the intra‐cellular metabolites. The endometabolome of the cells was determined by 1H‐NMR spectroscopy. Changes in specific metabolite concentrations upon intervention with δT were quantified with the Chenomx‐NMR Suite. MTS assay and FITC Annexin V stained flow cytometry analysis were performed to determine the anti‐proliferative effects and induction of apoptosis by T3. The expression of glutamine transporters and mTOR pathway proteins were assessed using western blot analysis. The data shows that T3 significantly inhibited glutamine uptake in H1299 cells. The inhibition of glutamine uptake into proliferating cells resulted in dose dependent inhibition of cell proliferation and induction of apoptosis in both cell lines. Further validation using western blot data exhibits inhibition of SLC7A5 glutamine transporter and mTOR pathway proteins (P‐mTOR, mTOR, P‐AKT, AKT, S6K, c‐MYC. MMP‐9, and Bcl‐XL) by T3 in a dose dependent manner. Our findings suggest that the anticancer effects of T3 may occur via downregulation of mTOR1 pathway induced by inhibition of glutamine transporters.
In vivo Assessment of Lutein Absorption in Infant Formula using Mono‐diglycerides (MDG) Based Emulsification Technology
Authors: Vurma Mustafa , DeMichele Stephen , Lee Dana , Tso Patrick
Published: November 2023

Previously we showed using the lymph fistula rat model that specific mixtures of MDG improved lymphatic lutein absorption by solubilizing lutein and facilitating gastrointestinal micelle formation compared to lutein suspended in triglyceride oils. The primary objective of this study was to determine manufacturing/formulation options to incorporate MDG and lutein into infant formula to retain the observed lutein absorption benefits. We compared using a conscious lymph fistula rat model, the intestinal uptake and lymphatic transport of lutein from five different liquid infant formula prototypes. Five groups of male adult Sprague Dawley rats (n= 4–7/group) were randomized to gastrically receive (~6.0 μg lutein per animal) one of the following infant formula prototypes containing ~2 mg/L lutein: (AET‐Control) lutein is premixed with safflower oil and then added to the oil phase; (AET‐1) lutein premixed with MDG and lecithin, and then added the oil phase; (AET‐2) lutein is premixed with MDG, and then added to the water phase; (AET‐3) lutein is premixed with MDG, and then added to the oil phase; and (AET‐4) lutein is premixed with MDG and choline chloride, and then added to water phase. Aliquots of lymph were taken at fasting and hourly for 8 hours for lutein and fatty acid analyses. Lutein premixed with MDG and added to oil phase directly (AET‐3) during processing did not improve lutein absorption as compared with AET‐Control. On the other hand, gastric delivery of AET‐1, AET‐2, and AET‐4 significantly improved lymphatic lutein output 2–8 hours after formula feeding versus AET‐Control. The percent increases in lutein absorption over the 8 hour period were the following: AET‐1; 142%; p<0.05, AET‐2; 121%; p<0.05, and AET‐4; 165%; p<0.05. In addition, there were significant increases in lymphatic fatty acid (docosahexaenoic, arachidonic, palmitic) outputs during the study period for formulas AET‐1 and AET‐2 as compared with AET‐Control. These data show that when lutein is premixed with MDG and lecithin (AET‐1) and added to the oil phase or lutein premixed with MDG (AET‐2) and choline (AET‐4) and added to the water phase, preserved the lutein absorption benefits previously observed with MDG ingredient research. MDG based emulsification technology has the potential to improve the absorption of lutein in infants.

Support or Funding Information

DK059630, Abbott Nutrition ZA78
Acute Bioavailability of (Poly)phenolic Content of Different Varieties of Vaccinium spp. in Ovariectomized Rats
Authors: Maiz Maria , Cladis Dennis P. , Lachcik Pamela J. , Janle Elsa M. , Lila Mary Ann , Ferruzzi Mario G. , Weaver Connie M.
Published: November 2023

Blueberries have a wide range of health promoting activities due to their high levels of (poly)phenols, including anthocyanins, flavonols, and phenolic acids. However, there are 1200 varieties of blueberries and their phenolic content varies greatly. Thus, the goal of this investigation was to characterize the total phenolic content in 9 varieties of Vaccinium spp. berries as well as determine whether there is a difference in their bioavailability after an acute dose in ovariectomized rats.

Nine varieties of berries (Ira, Legacy, Montgomery, Onslow, SHF2B1‐21:3, Sampson, Cranberry, Bilberry and two Wild blueberry composites from the years 2012 and 2013) were selected based on PCA analysis of their anthocyanidin content and sourced from the North Carolina State University Blueberry Genome Repository. Berries were freeze‐dried and phenolics were extracted with 2% formic acid 80:20 MeOH:H2O solvent and purified using solid phase extraction (SPE). Total phenolic content was determined via the Folin‐Ciocalteu assay and individual phenolics where identified and quantified by HPLC‐MS.

For the bioavailability study, eighty‐four Sprague‐Dawley rats were given an acute dose of 25mg of total phenolics/kg of body weight by gavage as a freeze‐dried blueberry powder solution (aq.) and serial blood draws and urine were collected over 8 hours. Plasma samples were extracted by SPE and analyzed by HPLC‐MS/MS for anthocyanin glycosides and flavonoid metabolites. Pharmacokinetic parameters (AUC, Cmax, and Tmax) were determined.

Bilberry and Wild blueberry composite 2013 have a significantly higher total phenolic content than the other varieties, while Cranberry and Sampson were the least concentrated. Montgomery and Ira varieties had the highest bioavailability for Me‐Quercetin‐glc and Me‐Myricetin‐glc metabolites, respectively, by one‐way ANOVA (p < 0.05). The flavan‐3‐ols, catechin‐glucuronide (glc), epicatechin‐glc and their respective methylated metabolites, were not significantly different between varieties of blueberries.

Total (poly)phenolic content and bioavailability of individual phenolics varied widely among blueberry varieties. This information will serve to inform future investigations in cells, animals, and clinical trials.

Support or Funding Information

National Center for Complimentary and Alternative Medicine
Resistant starch (RS) fermentation is prevented by low potency antibiotics (AB) neomycin and ampicillin
Authors: Guice Justin L , Carvajal‐Aldaz Diana G , Page Ryan C , Raggio Anne M , Martin Roy J , Husseneder Claudia , Durham Holiday A , Geaghan James P. , Pelkman Christine L , Marco Maria L , Janes Marlene , Coulon Diana , Keenan Michael J
Published: November 2023

In a previous study we reduced bacterial genera that ferment RS with AB treatment then stopped AB prior to feeding RS. Subsequent fermentation of RS was good even without a transplant of microbiota from other rats fed RS and prior reduced bacterial genera increased (EB14). We now report use of the same AB (neomycin 0.5 g/L and ampicillin 1g/L in drinking water) at the same time as feeding RS. Sprague Dawley rats were assigned into one of four groups and fed ad libitum for four weeks. The groups included: 1. positive control (high RS, low fat diet [LF], no AB), 2. negative control (no RS, LF, no AB); 3. LF, high RS, AB; and 4. high fat (HF), high RS, AB. We measured effects of fermentation: pH of cecal contents, empty cecum weight (ECW), short‐chain fatty acids (SCFA), glucagon‐like peptide 1 (GLP‐1), RS in cecal contents, four bacterial genera known to be involved in fermentation of RS and total bacterial numbers. HF group 4 was included to see if AB might improve fermentation of RS compared to no AB because Cani et al. (2008) had reported that AB reduced endotoxemia in diet‐induced obesity. AB reduced Lactobacillus spp. and Bifidobacterium spp., but not Bacteroides spp. or total bacteria. Empty cecum weights of groups given AB and positive control were greater than the negative control. RS increased acetate and butyrate as well as decreased pH of cecal contents and increased empty cecum weights; AB treatment in both LF and HF prevented these effects (p<0.05). AB also had double the amount of GLP‐1 in serum and double the amount of RS in cecal contents compared to positive control, and negative control had minimal amounts of both GLP‐1 and RS. Thus, it appears that half of the RS was not fermented in the positive control and AB eliminated fermentation of RS when given at the same time as feeding of RS. However, increased ECW and GLP‐1 are a concern for future use of AB for reduction of fermentation of RS for mechanistic studies.

Support or Funding Information

Funding: Ingredion Incorporated and LSU AgCenter.
Effects of curcumin metabolites on human colon adenocarcinoma cells
Authors: Cheng An‐Chin , Lee Ming‐Fen , Li Cheng‐Ta , Chang Kai‐Hsun , Pan Min‐Hsiung
Published: November 2023

Stepwise cancer development involves metastatic migration as well as uncontrolled cell growth. Cancer cells undergo migration and invasion which allow them to translocate within tissues. Tetrahydrocurcumin (THC), a major metabolite of curcumin, exhibits anti‐carcinogenesis and antioxidation in vivo. In the present study, we found that THC and two other metabolites of curcumin, tetrahydrodemethoxycurcumin (THDC) and tetrahydrobisdemethoxycurcumin (THBC), could reduce the survival ratio of human colon adenocarcinoma cells, COLO205, HCT116, and HT29, in a dose‐dependent manner. The results also indicated that the fewer the methoxy groups of the curcumim metabolites, the more the inhibitory effects. However, curcumin metabolites did not trigger apoptotic cell death in COLO205 cells significantly, whereas curcumin did. Our data also strongly suggest a role of THBC in inhibiting cellular migration in HCT116 cells. Taken together, these results demonstrated the anticancer efficacy of curcumin metabolites which provides a potential application basis for the prevention of human colon cancer.

Support or Funding Information

This study was supported by NSC102‐2320‐B‐309‐001‐MY3 to M‐F Lee and NSC100‐2313‐B‐309‐003 to A‐C Cheng.
The Effect of Oil Palm Phenolics (OPP) on Urinary Metabolomic Profile in Atherogenic Diet Induced Rat Model of Alzheimer's Disease (AD)
Authors: Wu Yan , Srirajavatsavai Vindhyaja , Sambanthamurthi Ravigadevi , Gupta Smiti Vaid
Published: November 2023

Alzheimer's disease (AD) is a neurodegenerative disorder, estimated to strike more than 4.7 million individuals older than 65 in the USA. Since oxidative stress is a key component driving AD, we hypothesized that OPP, a water soluble fraction extracted from oil palm fruit rich in phenolics with high antioxidant activity, may attenuate its progression. In this study, we used the metabolomic approach to investigate the effect of OPP on change in metabolism produced in high cholesterol diet induced model for AD. Thirty two male Brown Norway rats were randomly assigned to 4 groups: the isocaloric control diet (C), high cholesterol (2% cholesterol) (H), high cholesterol + OPP (5% OPP) (HP) and high cholesterol + curcumin (2% curcumin) (HC). Urine samples were collected once a month for 6 months. Water maze was performed monthly to evaluate cognitive behavior. β‐amyloid was measured at termination of study as a marker for AD using ELISA. Curcumin, a dietary polyphenolic compound has been shown to have benefit in AD and was used as a positive control. Proton nuclear magnetic resonance (1H NMR) spectra, acquired using 500 MHz magnet were processed using ACD labs software. The processed, digitized NMR spectral data was subjected to multivariate data analysis (SIMCA P+). Principal component analysis (PCA) and partial least squares‐discriminant analysis (PLS‐DA) score plots showed clear separation among all four groups indicating differences in their metabolomics profiles. OPLS regression analysis gave significant correlations between the urinary metabolomic profiles and escape latency using water maze (R2=0.78) and the β amyloid burden (R2=0.73). The metabolites responsible for the differences in the metabolomic profile between different groups were quantified using CHENOMX NMR metabolite database. Some metabolites from the tryptophan‐kynurenine‐quinolinic pathway were significantly higher in the cholesterol fed animals (H) as compared to the control (C). Treatment with curcumin (HC) or OPP (HP) brought down the concentration of these metabolites significantly and closer to the control levels. This pathway has been shown to be perturbed in neurodegenerative diseases. Taken together, our data indicates that dietary OPP or curcumin have potential benefit in AD. In addition, specific urinary metabolites may serve as non‐invasive biomarkers for progression of neurodegenerative diseases including AD.
Omega‐3 fatty acid‐rich fish oil supplementation prevents rosiglitazone‐induced osteopenia in insulin resistant C57BL/6 mice
Authors: Rahman Md Mizanur , Halade Ganesh , Williams Paul
Published: November 2023

Rosiglitazone (RSG) is a very effective insulin sensitizing and glucose‐lowering drug, however, associated with bone loss resulting in increased peripheral fractures. RSG‐induced bone loss occurs mainly due to increased bone resorption, and bone marrow adiposity. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)‐rich fish oil is known to attenuate both bone resorption and bone marrow adiposity. We hypothesize that co‐administration of EPA and DHA‐rich fish oil prevents RSG‐induced bone loss. In this report, we investigated the effect of co‐administration of fish oil on RSG‐induced bone loss in C57BL/6 mice and the mechanisms underlying this preventive effect. Twelve‐month‐old C57BL/6 female mice were fed iso‐caloric diet supplemented with corn oil (CO), fish oil (FO), RSG+CO and RSG+FO for 5 months. Bone mineral density (BMD) was analyzed by dual energy x‐ray absorptiometry (DXA). RSG treatment reduced BMD in both CO and FO groups. However, mice fed RSG+FO exhibited significantly higher levels of bone mineral density (BMD) in different bone regions as compared to CO alone or RSG+CO fed groups. Reduced activity of COX‐2, enhanced activity of alkaline phosphatase were observed in LPS treated bone marrow (BM) cells from RSG+FO treated mice as compared to RSG+CO group. LPS treated BM cells from RSG+FO treated mice also expressed lower levels of Cathepsin K and PPAR‐γ as compared to RSG+CO group. LPS‐stimulated BM and splenocytes from RSG+FO also produced significantly lower level of pro‐inflammatory cytokines, TNF‐alpha and IL‐6 and higher level of anti‐inflammatory cytokine, IL‐10 as compared to RSG+CO group. We further examined the effect of DHA and EPA with RSG on bone morphogenic protein‐2 (BMP‐2)‐induced osteogenesis and adipogenesis in C3H10T1/2 cells and RANKL‐induced osteoclastogenesis in RAW264.7 cells in‐vitro. DHA restored RSG‐induced reduction of osteoblast differentiation and RSG‐induced up‐regulation of adipocyte differentiation in C3H10T1/2 cells. DHA also inhibited RSG‐induced up‐regulation of osteoclast differentiation of RANKL treated RAW264.7 cells. In conclusion, we provide evidence that FO co‐administration with RSG may prevent RSG‐induced bone loss by inhibiting markers of inflammation, osteoclastogenesis and adipogenesis and by enhancing osteogenesis in bone microenvironment.

Support or Funding Information

This study was supported by NIH contract grant number AG034233 (M.R.)
Preclinical safety profile of a depleting antibody against CRTh2 for asthma: Well tolerated despite unexpected CRTh2 expression on vascular pericytes in the central nervous system and gastric mucosa
Authors: Austin Cary D. , Huang Tao , Sharma Neeraj , Liu Shannon , Solon Margaret , Reyes Arthur , Paul Sarah , Yee Angie , Tao Janet , Chalasani Sreedevi , Bien‐Ly Nga , Barck Kai , Carrano Richard A.D. , Wang Jianyong , Rangell Linda , Bremer Miere , Danilenko Dimitry M. , Katavolos Paula , Hotzel Isidro , Reif Karen , Rajapaksa Kathila S.
Published: November 2023

CRTh2 is expressed on immune cells that drive asthma pathophysiology. Current treatment options for severe asthma are inadequate and therapeutic antibody‐mediated depletion of CRTh2‐expressing cells represents a promising new therapeutic strategy. Here we report for the first time that CRTh2 is not only expressed on immune cells, but also on microvasculature in the central nervous system (CNS) and gastric mucosa in humans. Microvascular expression of CRTh2 raises a safety concern because a therapeutic anti‐CRTh2 antibody with enhanced depletion capacity could lead to vascular damage. To evaluate this safety risk, we characterized microvascular expression in human and in transgenic mice expressing human CRTh2 protein (hCRTh2.BAC.Tg) and found that CRTh2 is not localized to microvascular endothelium that is directly exposed to circulating therapeutic antibody, but rather, to pericytes that in the CNS are shielded from direct circulatory exposure by the blood‐brain barrier. Immunohistochemical visualization of an intravenously administered anti‐CRTh2 antibody in transgenic mice revealed localization to microvascular pericytes in the gastric mucosa but not in the CNS, suggesting the blood‐brain barrier effectively limits pericyte exposure to circulating therapeutic antibody in the CNS. Repeated dosing with a depleting anti‐CRTh2 antibody in hCRTh2.BAC.Tg mice revealed linear pharmacokinetics and no drug‐related adverse findings in any tissues, including the CNS and gastric mucosa, despite complete depletion of CRTh2 expressing circulating eosinophils and basophils. Collectively, these studies demonstrate that the likelihood of drug‐related CNS or gastrointestinal toxicity in humans treated with a therapeutic depleting anti‐CRTh2 antibody is low despite pericyte expression of CRTh2 in these tissues.
Inaccurate Quantitation of IgM Monoclonal Proteins: A cause for concern
Authors: Holmes Earle W. , Bahar Burak , Whisler Barbara M. , Gryzbac Margie , Kahn Stephen E.
Published: November 2023

The detection and quantification of monoclonal proteins (MCP) play an essential role in the diagnosis and management of plasma cell dyscrasias. As a follow‐up to an unusual case in which our SPEP method severely underestimated the size of an IgM monoclonal protein in a patient with a provisional diagnosis of amyloidosis, we investigated the linearity of the Sebia Hydrasys, Hydragel beta 1–2 method ( Sebia, Inc. Norcross, GA 30093) for the recovery of IgM(n=19) and IgG(n=9) MCP using the manufacturer's routine protocol both before and after pre‐dilution of the serum with saline (SAL) or a chaotropic diluent containing a mild reducing agent(DPD). MCP concentrations in SPEP patterns from amido black‐stained gels were quantitated by the routine Sebia protocol, and the recoveries of the albumin and MCP bands were directly measured as total RBG intensities by image analysis(IA) using Image Studio Lite, 4.0 (Li‐Cor, Lincoln NE 68504). Results are presented as mean (min to max). IA showed that the mean recoveries of the albumin and MCP bands of diluted samples containing IgM MCP were increased by factors of 3.0‐fold (1.6‐ to 10.6) and 4.1‐fold (1.5 to 24.6), respectively, over those measured in the undiluted samples. Moreover, the MCP/albumin recovery ratio was increased by 1.2‐fold (0.7 to 2.4) compared to the undiluted serum. This ratio was significantly increased compared to the ratio of 0.95‐fold (0.8 to 1.1) observed for the specimens that contained IgG MCP (p<0.001, KW). These results showed that the routine Sebia method, which specifies the analysis of undiluted serum, frequently under‐estimated the concentration of IgM MCP, possibly owing to flaws in the sample application and/or the protein staining steps of the procedure. Pre‐dilution with SAL prior to analysis increased IgM MCP concentrations by a mean of 9.4 % (0 to 22.0). Pre‐dilution with DPD provided similar increases except in one case in which the concentrations of two, relatively small, IgM MCP were increased by 22 and 52%. In addition to an under‐recovery of IgM MCP by SPEP, we found that band size estimates based on the quantitation of the total IgM concentration by rate nephelometry (Immage 800, Beckman Coulter Inc., Brea CA 92821) were 1.3‐ to 2.7‐fold greater than those measured by SPEP. Both of the major methods used for estimating the size of MCP appear to have analytical limitations that compromise their accuracy. While either method might be adequate for monitoring changes in tumor burden over time, the discrepancies we noted may be a source of confusion and error in scenarios in which fixed clinical decision thresholds based on MCP concentration are used as criteria for diagnosis, disease progression, and response to treatment.
Acetylcholine‐signaling inhibitors for lung cancer therapy
Authors: Dasgupta Piyali , Lau Jamie K , Brown Kathleen C , Rollyson William D , Stover Cody A , Rimoldi John M , Cutler Stephen J , Hardman W Elaine , Carpenter A Betts , Chen Yi C
Published: November 2023

Cigarette smoking is a major risk factor for the development of small cell lung cancer (SCLC) and non‐small cell lung cancer (NSCLC). Nicotine is the addictive component of cigarette smoke. Nicotine (at concentrations present in the plasma of moderate smokers) accelerates the growth, angiogenesis and metastasis of lung cancers. The biological activity of nicotine is mediated by nicotinic acetylcholine receptors (nAChRs). The endogenous ligand for nAChRs in the lung is the neurotransmitter acetylcholine (ACh). ACh is known to be an autocrine growth factor for both SCLC and NSCLC. One of the ways that nicotine and cigarette smoke promote the growth of lung cancers is by promoting the production of ACh, which binds to its cognate receptor (on lung cancer cells) in an autocrine manner and induces the proliferation of lung cancer cells. The objective of the present study is to investigate the growth‐inhibitory activity of acetylcholine‐signaling pathway in SCLC and NSCLC. We show that three acetylcholine‐signaling inhibitors, BW813U, hemicholinium‐3 and vesamicol, cause robust apoptosis in human NSCLCs. The anti‐cancer activity of vesamicol was also confirmed in athymic mouse models. Although there are several targeted therapies against NSCLC, they do not provide optimum efficacy in NSCLC patients who are exposed to cigarette smoke. Our studies will facilitate the identification of targeted therapies for the population of lung cancer patients who are smokers.

Support or Funding Information

Funding for our study was supported by an NIH R15‐AREA Grant (1R15HL113681‐01A1), ACTSI Grant from Marshall University, the Flight Attendant Medical Research Institute YCSA Grant (82115), and the West Virginia IDeA Network of Biomedical Research Excellence (P20RR016477 and P20GM103434).
Role of the transcription factor Yin Yang 1 in non‐small cell lung cancer
Authors: BOUCHERAT Olivier , CHABOT Sophie , BOURGEOIS Alice , PROVENCHER Steeve , PAULIN Roxane , MALTAIS François , BONNET Sébastien
Published: November 2023

Background

Lung cancer is the most prevalent cancer type worldwide by both diagnosis and mortality, with non‐small cell lung cancers (NSCLCs) featuring as the most common histological subtype, the latter of which consists of squamous (epidermoid), adenocarcinoma, and large cell carcinoma. Rates of lung cancer will continue to rise owing to high levels of cigarette consumption and aging populations. Despite increased surgical resection rates, optimization of chemo‐radiotherapy and the introduction of targeted therapies, the treatment outcome for NSCLC remains poor. Therefore, identification of new molecular targets, which are essential for proliferation of tumor cells, will benefit both treatment and chemoprevention of NSCLC. The Yin Yang 1 (YY1) transcription factor has a pivotal role in normal biological processes such as development, differentiation, replication and cell proliferation exerting its effects on a huge number of genes involved in these processes. Due to its functions during lung morphogenesis, we hypothesize that YY1 is overexpressed and contributes to the pro‐proliferative and anti‐apoptotic phenotype of cancerous cells.

Method/Results

Expression level of YY1 was first examined by immunohistochemistry (n=5) and western blot (n=11) in resected tumor tissue of NSCLC patients (stages I–III) and compared to adjacent non‐cancerous lung tissue. Results showed that YY1 expression is significantly increased in squamous cell carcinoma and adenocarcinoma of the lung. A greater proliferation rate (Ki67 labeling) was associated with strong YY1 expression in these human lung tumors. In vitro, transient transfection experiments with A549 lung adenocarcinoma cells showed that depletion of YY1 levels by short interfering RNA reduced proliferation (Ki67 labeling and MTT assay, p<0,001), resistance to apoptosis (Annexin V assay, cleaved caspase‐3 expression, p<0,05) and migration (wound healing assay, p<0,001). In addition, YY1 knockdown improved in vitro NSCLC sensitivity to Etoposide/Cisplatin, two chemotherapeutic agents.

Conclusion

Taken together, these findings suggest that up‐regulation of YY1 plays an important role in the development of lung cancer. Susceptibility of YY1‐depleted A549 cells and YY1 heterozygous mice to induce tumors is currently under investigation. Analysis of YY1 expression in a larger cohort of patients may reveal prognostic information and deserves further study.

Support or Funding Information

None
NLRP3 Inflammasome as a Novel Target for Docosahexaenoic Acid and Its Metabolites to Abrogate Glomerular Injury during Hyperhomocysteinemia
Authors: Chen Zhida , Conley Sabena M. , Li Guangbi , Xia Min , Gehr Todd W. , Boini Krishna M. , Li Pin‐Lan
Published: November 2023

It has been reported that Ω‐3 fatty acid, docosahexaenoic acid (DHA) is a clinically significant therapeutic aimed at reducing inflammation and prevention of inflammatory injury in glomeruli associated with different diseases such as hypertension and diabetes mellitus. However, the mechanism by which DHA and its metabolite exert anti‐inflammatory actions remains unclear. The present study was designed to test whether the intracellular, NOD‐like receptor family, pyrin domain containing 3 (NLRP3) inflammasome may serve as a novel therapeutic target for treatment or prevention of glomerular injury in hyperhomocysteinemia (hHcys). To answer this question, Nlrp3 knockout (Nlrp3−/−) and wild type (Nlrp3+/+) mice were fed a folate‐free (FF) or normal diet (ND) for 4 weeks to induce hHcys. Our studies found that using this experimental model of hHcys doubled plasma Hcys levels in FF fed mice. Neither administration of DHA nor a lipoxygenase (LOX) inhibitor, cinnamyl‐3, 4‐dihy‐droxyl‐cyanocinnamate (CDC) had an effect on Hcys levels. Confocal microscopic analysis showed that the FF diet increased the colocalization of inflammasome protein NLRP3 with ASC or with caspase‐1, suggesting NLRP3 inflammasome formation during hHcys induced by the FF diet. DHA treatment almost completely blocked this formation of NLRP3 inflammasome in glomeruli. However, mice receiving both DHA and CDC, showed enhanced glomerular inflammasome formation during hHcys. By immunohistochemical staining, we revealed that treatment with DHA and CDC inhibited IL‐1β production in glomeruli of mice with hHcys. Furthermore, the urinary protein and albumin excretion as well as glomerular damage index were significantly lower in mice with hHcys receiving DHA alone, but not in mice treated with DHA and CDC. These results provide the first evidence that DHA and its lipoxygenase products may abrogate the activation of NLRP3 inflammasomes and thereby prevent glomerular injury or sclerosis induced by hHcys. The NLRP3 inflammasome in podocytes of glomeruli may be a potential new therapeutic target for treatment of chronic glomerular disease by DHA or its metabolites.

Support or Funding Information

Supported by NIH grants DK054927,HL07356 & HL057244