bluuebunny/crossref_metadata_2025 huggingface dataset:
The Randomization to Randomization (R2R) Method: an RCT application with Caffeine
Authors: Jeansonne Madeline M , Parman Mariel A , Idigo Adeniyi J , George Brandon J , Fontaine Kevin R , Dutton Gareth R , Soleymani Taraneh , Allison David B
Published: November 2023

Background

In double‐blind, placebo‐controlled RCTs, patients do not know for certain whether they are receiving the active treatment. However, when a doctor prescribes a treatment, a patient is certain of receiving the active treatment. Thus, traditional RCTs do not test the active treatment under real‐life conditions because they do not account for potential interactions between expectancy effects and treatment on outcomes.

Objective

To estimate treatment effects free from expectancy effects and determine if expectancy effects interact with treatment.

Methods

After an overnight fast from caffeine, participants were randomized to a probability of receiving 200mg caffeine versus placebo pills (probabilities ranged from 0.1 to 0.9 in 0.1 increments). Expectancies were manipulated by revealing the randomization probability to the participant at the beginning of the study visit. Participants were then randomized to caffeine or placebo based on assigned probability, consumed their pills and, after 30 minutes, completed mood and vigilance testing. Generalized linear models with appropriate outcome distributions were used to analyze the data.

Results

A significant interaction (p=0.0054) was observed between the effects of caffeine and assigned probability on the vigor‐activity mood cluster, such that increased probability of receiving the caffeine produced higher scores of positive and/or energetic feelings for the caffeine group only. The estimate of the effect of caffeine when the probability was 1 (7.9, 95% CI: 3.3,12.6) differed from the estimate from those assigned a probability of 0.5 (2.9, 95% CI: −1.1,6.7) as in a standard RCT. Those who received caffeine had shorter reaction times to the vigilance stimuli (p=0.001) and greater odds (OR=1.774, 95% CI: 1.381, 2.281) of correctly detecting stimuli during the vigilance test (p<0.0001). An expectancy effect was found (p=0.0399) such that increased probability of receiving the caffeine decreased the number of stimuli correctly detected during the vigilance test.

Conclusions

This was a proof of concept study. Results are preliminary but suggest that the R2R design is feasible and capable of detecting treatment by expectancy interactions without deceiving participants. Furthermore, it is possible that failing to account for this interaction can result in conclusions that are not representative of effects under actual conditions of use (i.e. when the probability is 1).

Support or Funding Information

Funded by internal university funds.
Biochemical Comparison of AAC1 Enriched Chicken Broth to Homemade Chicken Broth: Potential Benefits to Joint Health
Authors: Durham Paul , Hawkins Jordan L
Published: November 2023

Objective

To investigate the differences in the level or activity of biochemical molecules contained in enriched AAC1 and traditional homemade chicken broth that are implicated in improving and maintaining good joint health.

Background

Chicken broth is a complex mixture containing extractives obtained from cooking of chicken parts or whole chickens. The method used to prepare the chicken broth can significantly alter the composition of the broth. Different chicken broths are likely to exhibit diverse compositions which may help to explain the varying nutritional and health benefits for different broths. The commercially available AAC1 both is prepared using a different process than traditional homemade chicken broth and thus, the biochemical properties would be expected to be distinctive.

Methods

AAC1 and homemade chicken broths were dissolved in water to a final solids concentration (w/v) of 1%. The amount of hyrdoxyproline was determined by LC‐MS and chondroitin sulfate using enzymatic digestion and LC‐UV detection. The level of polyphenolic compounds was quantified using a modified Folin‐Ciocalteau method, while the activity of COX‐1 and COX‐2 enzymes was assessed using a commercially available COX inhibitor assay. All data are reported as mean ± standard error of the mean (significance at p < 0.05).

Results

The AAC1 broth was found to contain a higher percentage of collagen based on the levels of hydroxyproline (at least 8% hydroxyproline by weight of total amino acids in the composition) when compared to homemade broth. Similarly, AAC1 contained a higher amount of chondroitin sulfate (8.797% w/w) compared to homemade. While both chicken broths contained polyphenols, AAC1 broth exhibited approximately 7 times more Gallic acid equivalents (4828.8 μg/mL GAE) than homemade broth. Interestingly, AAC1 had a greater ratio of COX‐2 inhibition compared to COX‐1 inhibition (18.34) with over 40% inhibition of COX‐2 enzyme activity.

Conclusions

When compared to homemade chicken broth, we found that AAC1 broth is enriched in biological molecules associated with joint health and decreasing inflammation. Thus, our findings support the notion that dietary inclusion of AAC1 should be beneficial in promoting and maintaining healthy joints.

Support or Funding Information

International Dehydrated Food
In vitro Antioxidant Activity and α‐Glucosidases inhibitory potential of Amadori Rearrangement Compound, Lysinyl‐fructose (LF)
Authors: Choi Hwang‐Yong , Jo Sung‐Hoon , Ha Kyoung‐Soo , Jang Hae‐Dong , Kwon Young‐In
Published: November 2023

Lysine and glucose‐containing food ingredients, a new amadori reaction compound lysinyl‐fructose (LF) was synthesizd and purified. Amino acid derivatives such as LF are formed through Amadori rearrangement, the early step of Maillard reaction from lysine and glucose. Millard reaction products were prepared by heating of Lysine and glucose at 70 °C for 3 hours. After cooling to a certain temperature and the reaction solution was heat treated for 10 minutes centrifuged (10,000 × g), and concentrated using a vacuum rotary evaporator taking the supernatant. The sample was concentrated and purified by cation exchange resin to obtain a purified product. To confirm the structure and purity of the sample, LC/MS/MS and HPLC systems were performed. In order to evaluate the bioactivities of LF, antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) method. The inhibitory activity of LF against porcine pancreatic α‐amylase and rat intestinal α‐glucosidases(sucrase, maltase, glucoamylase) was also investigated. LF had a high α‐glucosidase inhibitory activity as well as the antioxidant activity. These results might be helpful to understand the action activities mechanism of LF.
Dairy food intake and perceptions 1995 to 2009: is it time to promote dairy food consumption?
Authors: Kerr Deborah Anne , Meng Xingqiong , Devine Amanda , Prince Richard L , Pollard Christina M
Published: November 2023

Background

There is evidence for the health benefits of dairy food consumption throughout the lifecycle. Government dietary guidelines advise the population to eat a wide variety of nutritious foods including foods from the dairy food group, particularly lower fat versions, to promote health and protect against disease. Knowledge, attitudes and intentions are key to adherence to dietary recommendations.

Objective

To assess trends in self‐reported dairy food consumption and intentions regarding these foods among Western Australia adults 18 to 64 years.

Methods

Data were pooled from the 1995, 1998, 2001, 2004, 2009, and 2012 Health Department's Nutrition Monitoring Survey Series of 7,044 adults. Outcome measures included type and amount of dairy foods eaten on the day prior. Sociodemographic characteristics, perceptions of current dairy food intake and intentions to change were measured. Multinominal logistic regression models were used to assess the factors associated with consumption.

Results

Most respondents (96%) reported consuming dairy products (milk, yoghurt, and/or cheese) on the day prior to the survey. Although the proportion consuming dairy foods remained relatively stable across survey years, the amount consumed decreased from 2.7 (95% CI: 2.6, 2.9) serves/d 1995 to 2.2 (95% CI: 2.1, 2.3) serves/d in 2012. Men consumed more serves of dairy compared with women (Coef 0.38 95% CI: 0.27–0.49; p<0.001). The amount consumed decreased with age (Coef −0.57 95% CI: −0.80,−0.34; p<0.001) for 55–64 years compared to 18–24 years. Overweight or obese people consumed fewer dairy foods serves than those of a healthy body weight (Coef −0.26, 95% CI: −0.37, −0.14; p<0.001). Respondents were twice as likely to be ‘thinking about trying to change their intake’ compared to ‘actually trying to increase consumption’ in 2012 compared with 1995 (RRR 2.26, CI 1.36, 3.74; p<0.01). Men were more likely than women to have tried to decrease their dairy food intake in the past year (RRR 2.08, 95% CI: 1.35, 3.22; p<0.001). The perception that ‘I already eat enough dairy foods’ increased as a barrier to eating more, from 64.5%, 95% CI: 61.2–67.6 in 1995 to 77.1%, 95% CI: 74.3–79.6 in 2012 (p<0.001).

Conclusions

These findings indicate that many adults believe they already eat enough dairy foods yet their intake is less than recommended and is decreasing over time. Health promotion interventions to address the declining consumption of dairy foods as part of a healthy dietary pattern are warranted and should challenge this misconception.

Support or Funding Information

Healthway funded Curtin University's Food Law, Policy and Communications to Improve Public Health Project to assist the translation of research into practice
Effect of Low‐Glycemic Index/Load Diets on Body Weight: A Systematic Review and Meta‐Analysis
Authors: Braunstein Catherine Rose , Mejia Sonia Blanco , Stoiko Elena , Noronha Jarvis C , Choo Vivian , Ha Vanessa , Augustin Livia , Jenkins David JA , Kendall Cyril WC , Sievenpiper John L
Published: November 2023

Objective

It is unclear whether low glycemic index and load diets contribute to weight loss. To synthesize the evidence of the effect of low glycemic index/load (GI/GL) diets on body weight in order to inform clinical practice guidelines, we conducted a systematic review and meta‐analysis of randomized controlled trials.

Methods

We searched Medline, EMBASE, and the Cochrane Library (through June 2 2015). We included randomized controlled trials ≥12 weeks investigating the effect of a low‐GI or low‐GL intervention compared with an isocaloric control diet on body weight in participants that are overweight or obese (BMI >25 kg/m2). Two independent reviewers extracted all relevant data, assessed risk of bias of individual trials using the Cochrane Risk of Bias Tool. Data were pooled using the generic inverse variance method and expressed as mean differences (MD) with 95% confidence intervals (CIs). Heterogeneity was assessed by the Cochran Q statistic and quantified by the I2 statistic.

Results

Eligibility criteria were met by 19 randomized controlled trials (RCTs) (22 comparisons) involving 1,577 overweight or obese participants. Low GI/GL diets resulted in a non‐significant decrease in body weight compared with higher GI/GL control diets (MD= −0.32 kg (95% CIs −0.86 kg, 0.23 kg) with evidence of substantial inter‐study heterogeneity (I2=57%, p<0.001).

Limitations

Most of the trials were of moderate duration (<6 months) and there was substantial unexplained inter‐study heterogeneity.

Conclusion

Low GI/GL diets do not lead to significantly more weight loss than higher GI/GL diets over the moderate term in overweight or obese people. To address the sources of uncertainty, there is a need for larger, longer, higher quality trials.

Support or Funding Information

Financial support for this work given by The Canadian Institutes of Health Research (funding reference number, 129920) through the Canada‐wide Human Nutrition Trialists’ Network (NTN), the PSI foundation, and the Canadian Diabetes Association.
A dietary intervention of Fish oil can restore metabolic dysfunction and improve estrous cycle irregularity in obese C57BL/6 female mice
Authors: Godwin Simone Marie , Beech LeeAnna , Buddington Randy , van der Merwe Marie
Published: November 2023

Reproduction, an energetically costly process, is subject to nutritional and metabolic control. There is a high occurrence of female infertility in obese women, and Polycystic Ovarian Syndrome (PCOS), a condition characterized by androgen excess and oligo‐ and amenorrhea, is highly associated with obesity and diabetes mellitus. Obesity is characterized by chronic, low‐grade inflammation, and increased circulating pro‐inflammatory cytokines. These pro‐inflammatory molecules can alter insulin signaling, thereby contributing to glucose intolerance and insulin resistance, both hallmarks of type 2 diabetes. This study seeks to elucidate the relationship between obesity, inflammation, and reproductive dysfunction through diet intervention. Diets rich in n‐3 poly‐unsaturated fatty acids (PUFAs), such as is found in fish oil, are considered to have anti‐inflammatory effects. We induced obesity and reproductive dysfunction in C57BL/6 female mice by feeding a “Western‐type” high fat diet (45% energy from fat) with lard as the source of fat. At 16 weeks of age, the experimental group was switched to a diet of identical composition but with fish oil as a source of fat for an additional 8 weeks. Changes in estrous cycle regularity were assessed by vaginal cytology. Metabolic outcomes were evaluated by measuring body weight, food consumption, glucose tolerance, fasting insulin levels, and adipose and liver histology. Inflammatory status was determined by circulating cytokine and adipokine levels and changes in regulatory immune cell populations. Reproductive irregularity was successfully established by 16 weeks of age at which time the mice also had increased glucose intolerance and higher circulating Leptin levels. At 24 weeks, mice that had remained on the lard based diet weighed more, had a dramatic increase in fat deposition in the liver, showed worse glucose tolerance response, had higher fasting insulin levels than their fish‐oil counterparts, but did not have increased systemic pro‐inflammatory cytokine levels. Consumption of the fish oil diet for 8 weeks did not restore estrous cycle regularity for all C57BL/6 female mice, however, metabolic parameters dramatically improved during the course of intervention. These findings suggest including fish oil in the diet of obese females will improve metabolic status, but a longer period of consumption may be needed to restore normal reproductive functions.

Support or Funding Information

Funded by Start Up funds from the University of Memphis
Groundsel Bush (Baccharis halimifolia) Enhances Adipocyte Development and Attenuates Inflammation in Adipocytes in vitro
Authors: Fuller Scott Eric , Boudreau Anik L , Stephens Jacqueline M
Published: November 2023

Introduction

Adipocytes serve a critical function not only as a reservoir of excess caloric energy but also as an endocrine organ regulating metabolic homeostasis. Therefore, interventions aimed at enhancing adipocyte development and function have the potential to ameliorate conditions associated with metabolic syndrome, such as ectopic fat accumulation and insulin resistance. Here we report evidence that a novel botanical, Baccharis halimifolia (groundsel bush), enhances adipocyte development and function in vitro.

Methods

For in vitro assessment of adipocyte development, differentiating adipocytes were treated with groundsel bush (GB) extract at three doses (5, 20, and 50 μg/mL) and the effects of GB were assessed by performing qPCR and immunoblotting to determine expression of adipogenic gene and protein expression, respectively. Neutral lipid accumulation in differentiating adipocytes was measured by Bodipy or Oil Red O staining. In mature murine adipocytes, adiponectin protein expression was assessed by western blot. The effectiveness of GB in attenuating inflammatory responses in mature adipocytes and RAW macrophages was evaluated by western blot analysis of adiponectin protein expression in TNFα‐treated adipocytes and by analysis of inflammatory gene expression in LPS‐treated RAW macrophages.

Results

In differentiating adipocytes, GB treatment enhanced the expression of adipogenic marker genes and protein expression of adiponectin and aP2, and also increased lipid accumulation. In mature adipocytes, GB increased gene and protein expression of adiponectin while decreasing resistin gene expression. GB attenuated TNFα‐induced repression of adiponectin protein expression in mature adipocytes while also reducing gene expression of MCP‐1 in LPS‐treated RAW macrophages.

Conclusions

The present study provides evidence that GB enhances adipocyte differentiation and neutral lipid accumulation, while also increasing adiponectin expression in mature adipocytes. GB also attenuates inflammatory responses in adipocytes and RAW macrophages. Collectively, these results indicate that GB has the potential to favorably modulate adipocyte function and therefore merits further study.

Support or Funding Information

This work was supported by NIH P50 AT002776. SEF was supported by institutional training grant NIH T32 AT004094 awarded to PBRC.
Dietary raspberries ameliorate metabolic syndromes in diet‐induced obese mice
Authors: Zhu Mei‐Jun , Kang Yifei , García Melisa Paola González , Liang Xingwei , Xue Yansong , Pan Hong , Du Min
Published: November 2023

Introduction

Obesity affects more than one third of the US population, which predisposes to a number of serious diseases. The typical “Western diet” is mainly blamed for the current obesity epidemics. Thus, optimizing the diet composition will be one of the primary approaches to prevent obesity. Raspberry contains very high amounts of dietary fibers and polyphenolic compounds, which are known for their anti‐oxidative and anti‐inflammatory effects, and also their likely effects to prevent obesity.

Objective

This study was to evaluate beneficial effects of raspberry supplementation on adiposity, glucose and lipid homeostasis impaired due to diet‐induced obesity.

Methods

Wild‐type male Mice (six weeks old) were randomized into 4 groups receiving a control or typical western diet (HFD) supplemented with or without 5% freeze dried raspberry for 12 weeks, when mice were sacrificed for tissue collection.

Results

Feed intake was not different among dietary groups. HFD feeding dramatically increased body weight, which can be alleviated by raspberry supplementation towards to the end of feeding trail. In addition, baseline blood glucose is significantly elevated in HFD group, which were reduced by raspberry supplementation. Addition of raspberry improved glucose tolerance and insulin sensitivity impaired by westernized HFD, and has a tendency to reduce adiposity elicited by HFD. In addition, Dietary Raspberry reduced abundance of lipid droplets in the liver of HFD mice as indicated by Oil‐Red‐O staining and has a tendency to decrease serum triglyceride level in HFD mice.

Conclusion

Dietary raspberry supplementation reduced adiposity and improved glucose and lipid homeostasis impaired by diet‐induced obesity.

Support or Funding Information

NATL PROC RASPBERRY CNCL
Novel mechanism responsible for regulation of branched‐chain alpha‐ketoacid dehydrogenase kinase
Authors: Shimomura Yoshiharu , Kondo Yusuke , Ito Rina , Tsuji Ai , Shibata Katsumi , Kitaura Yasuyuki
Published: November 2023

Branched‐chain amino acids (BCAAs: leucine, isoleucine, and valine) are indispensable amino acids. The BCAA catabolism is regulated by branched‐chain alpha‐ketoacid dehydrogenase (BCKDH), which is located at the second step of the mitochondrial catabolic pathway. The BCKDH is regulated by a phosphorylation‐dephosphorylation cycle, and BCKDH kinase (BDK) is responsible for inactivation of the BCKDH by phosphorylation of the E1‐alpha component. It has been demonstrated that two forms of BDK exist in rat liver mitochondria: a form bound to BCKDH and a free form. Only the bound form is suggested to be active. In the present study, we found a BDK‐inactivating factor in mitochondrial extracts from rat liver. The BDK‐inactivating factor was separated from the BCKDH‐BDK complex by high‐speed centrifugation and both are reconstitutable. We here report the characteristics of the BDK‐inactivating factor, which released the BDK from the BCKDH in a Ca ion‐dependent manner. From these findings, we propose the novel mechanism responsible for the regulation of BCAA catabolism.

Support or Funding Information

Funding for this research was in part received from the AJINOMOTO AMINO ACID RESEARCH PROGRAM.
Genes and sequence variants in weight loss in response to diet and exercise: Meta‐analysis
Authors: Jung Yanghoon Peter , Sanchez Brittany , Kreider Richard
Published: November 2023

BACKGROUND

Causative factors to obesity are related to diet and sedentary lifestyle; as well as biology, genomic variation and epigenetic events. A plethora of suggested modified behaviors to control and reduce body weight include nutritional intervention, exercise programming, behavioral or psychological approaches, pharmacological prescriptions, and even surgical procedures. Although a standardized intervention may be shown to be effective, high variability still exists in regards to individual response for controlled behavioral change. Many additional causative factors for this fluctuation remain unknown. A clear understanding of key gene‐diet interactions, in particular single nucleotide polymorphisms (SNPs), will aid in implementing behavioral modifications more effectively, based on individual genetic predisposition in humans.

PURPOSE

To define SNPs that impact weight loss, weight‐maintenance through diet, exercise, or lifestyle intervention.

METHODS

A critical review of 43 randomized controlled trials regarding SNPs and their regulatory role in weight loss was conducted. Databases searched were NCBI PubMed, Google Scholar, and Web of Science. Only controlled intervention trials were selected for review. A total of 9 independent studies with 2140 subjects (1103 with genotype FTO rs9939609, 269 with ADRB3 rs4994, 495 with AdipoQ rs2241766, and 273 with FABP2 rs1799883) were included. The criterions for the examined studies were weight loss induced by diet and exercise, or lifestyle modifications. Meta‐analysis was performed by Review Manager (RevMan) 5.2 (Cochrane, UK).

RESULTS

The meta‐analysis revealed that the carrier of FTO rs9939609 A‐allele had significantly higher weight loss than the non‐carriers (mean difference = 0.70, 95% CI (0.16, 1.24), p = 0.01, pheterogeneity = 0.89). There were no significant findings of ADRB3 rs4994 (mean difference = −0.42, 95% CI (−4.03, 3.19), p = 0.23, pheterogeneity = 0.76), FABP2 rs1799883 (mean difference = 0.13, 95% CI (−0.23, 0.49), p = 0.47, pheterogeneity = 0.95), and FABP2 rs1799883 (mean difference = −0.89, 95% CI (−4.69, 2.90), p = 0.67, pheterogeneity = 0.81)

CONCLUSION

FTO rs9939609 A‐allele carriers are more favorable to weight loss in response to diet and exercise intervention.
A Comparison of Effects of Lard and Hydrogenated Vegetable Shortening on the Development of High‐Fat Diet‐Induced Obesity in Rats
Authors: Kubant Ruslan , Poon Abraham N , Sanchez‐Hernandez Diana , Domenichiello Anthony F , Huot Pedro SP , Pannia Emanuela , Cho Clara E , Hunschede Sascha , Bazinet Richard P , Anderson G. Harvey
Published: November 2023

Background

Obesity is associated with increased consumption and preference for dietary fat. Experimental models of fat‐induced obesity use either lard or vegetable shortening. Yet, there are no direct comparisons of these commonly used fat sources, or the influence of their fatty acid composition on the development of diet‐induced obesity.

Objective

To compare the effects of lard and hydrogenated vegetable shortening diets, which differ in their fatty acid composition, on weight gain and the development of obesity and insulin resistance in rats.

Methods and Design

Male Wistar rats were fed ad libitum for 14 weeks high‐fat diets containing either: 1) high vegetable fat (HVF, 60 kcal% from vegetable shortening), 2) high lard fat (HLF, 60 kcal% from lard) or regular fat (control, 16 kcal% from vegetable shortening). Body weight, food intake, adipose tissue mass, serum glucose, insulin and fatty acid composition of diets were measured.

Results

Rats fed either of the two high fat diets had higher energy intake, weight gain and fat accretion than rats fed the regular fat diet. However, rats fed the HLF diet consumed more calories and gained more weight and body fat with greater increases of 32% in total (158.5±8.2 vs 120.2±6.6 g, P<0.05), 30% in visceral (104.4±5.2 vs 80.3±4.2 g, P<0.05) and 36% in subcutaneous fat mass (54.1±3.6 vs 39.9±3.1 g, P<0.05), compared to rats fed the HVF diet. Higher visceral adiposity was positively correlated with serum insulin (r2=0.376, P<0.05) and HOMA‐IR (r2=0.391, P<0.05).

Conclusion

We conclude that lard based high‐fat diets accentuate the increase in weight gain and the development of obesity and insulin resistance more than vegetable shortening diets. These results further point to the importance of describing fatty acid composition and type of fat used in determining outcomes of consuming high‐fat diets.

Support or Funding Information

Canadian Institute of Health Research, Institute of Nutrition, Metabolism and Diabetes (CIHR‐INMD), Reference MOP‐130286.
The Effect of Feeding Soy Protein Isolate and/or Omega‐3 Polyunsaturated Fatty Acids on the Spleen of Female PCK Rats
Authors: Gibson Lauren B. , Maditz Kaitlin , Benedito Vagner A , Tou Janet C
Published: November 2023

Few studies have investigated the effect of diet on the spleen. Studies have suggested that spleen lipid metabolism may play a role in non‐alcoholic fatty liver disease (NAFLD). Upregulated de novo lipogenesis (DNL) in the liver can cause NAFLD that can progress to cirrhosis and liver failure. This has important implications with NAFLD being the leading cause of chronic liver disease in adults and children consuming Westernized diets. The objective of this study was to determine the effect of diet on splenic DNL and inflammatory response. Female PCK rats aged 28 days were randomly assigned to one of four purified diets (n=12/group) and fed for 12 weeks. Diet groups consisted of 1) Casein + Corn Oil (Casein + CO), 2) Casein + Soybean Oil (Casein + SO), 3) Soy protein Isolate + Soybean Oil (SPI + SO), or 4) Soy protein Isolate + a 1:1 soybean: salmon oil blend (SPI + SB). Previously, we found NAFLD to be highest (P=0.003) in the SPI+SB group. In this study, the SPI+SB group showed a trend of high splenic weight (P=0.10). Spleen length was longer (P=0.02) in the SPI+SB fed group compared to the Casein + CO and Casein + SO groups. Splenic fatty acid profiles were determined by gas chromatography. Splenic content of saturated fatty acids (palmitic acid (C16:0) and stearic acid (C18:0)) and monounsaturated fatty acids (palmitoleic acid (C16:1n‐7) and oleic acid (C18:1n‐9)) were not significantly different between diet groups. Gene expression by real time quantitative polymerase chain reaction (RT‐qPCR) determined no significant difference in expression of transcription factors regulating DNL in the spleen including carbohydrate‐responsive element binding protein (ChREBP) and sterol regulatory element binding protein 1 (SREBP‐1c). The expression of transcription factor peroxisome proliferator‐activated receptor alpha (PPAR‐a) gene was used as a marker of splenic lipolysis, but showed no significant differences among diet groups. Rats fed the SPI+SB diet containing the highest (P<0.001) dietary eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA) exhibited the highest (P<0.001) splenic EPA and DHA, and the lowest (P=0.03) splenic content of AA. However, no significant differences in the expression of genes involved in regulating inflammation (tumor necrosis factor alpha (TNF‐a), interleukin‐6 (IL‐6), and interleukin 10 (IL‐10)) existed between diet groups. Based on these results, the diet treatments had no significant effect on splenic lipid metabolism or inflammatory function in PCK rats with NAFLD.

Support or Funding Information

This project was supported by the United Soybean Board Soybean Health Incentive Grant, West Virginia University PSCoR 10008736.6‐11‐NT10055R, West Virginia University Agriculture and Forestry Experimental Station Hatch Grant WVAA00665, and West Virginia University summer undergraduate education program.
High Maternal Vitamin D Status in utero through Weaning Beneficially Affects the Gut Microbiota‐Bone Axis in Obese Male Mice
Authors: Villa Christopher Ross , Comelli Elena M. , Taibi Amel , Chen Jianmin , Sacco Sandra , Ward Wendy E.
Published: November 2023

The gut microbiota may impact bone health via its released lipopolysaccharides (LPS), peptidoglycans or other metabolites, which we named the gut microbiota‐bone axis. A high‐fat high‐sugar diet increases serum LPS, decreases gut Bacteroides counts and weakens bones. In contrast, vitamin D has been recently shown to positively influence the composition of the gut microbiota, in addition to its important role in bone health. Interestingly, epidemiological studies show that prenatal vitamin D status is a critical contributor to offspring bone status. The purpose of this study was to investigate if exposure to high dietary vitamin D in utero and through weaning positively impacts gut microbiota and protects against chronic inflammation and compromised bone health in adult male mice fed a high‐fat high‐sugar diet.

C57BL/6J dams were fed modified AIN93G containing high (H, 5000 IU/kg diet) or low (L, 25 IU/kg diet) vitamin D during pregnancy and lactation. At weaning, male pups remained on their respective vitamin D levels (HH or LL) or were switched (HL or LH) and fed a high‐fat high‐sugar diet to induce chronic inflammation. At seven months of age, colonic Bacteroides and serum LPS were quantified by quantitative PCR and ELISA and bone microarchitecture at the distal femur and lumbar vertebra 2 was assessed via micro computed tomography.

Higher maternal but not offspring vitamin D status resulted in higher colonic Bacteroides counts (p=0.0004) and lower serum LPS (p=0.011) such that Bacteroides counts were negatively correlated with serum LPS (Spearman r=−0.49; p=0.018). Femur microarchitecture of offspring was also improved through higher maternal vitamin D status. Specifically, higher maternal vitamin D status resulted in offspring with higher bone volume/tissue volume (p=0.052), higher trabecular number (p=0.022) and lower trabecular separation (p=0.015) at distal femur and higher bone volume/tissue volume (p=0.049) at lumbar vertebra 2 compared to mothers with low vitamin D status. This suggests that the gut microbiota‐bone axis is programmed in utero and through weaning and is susceptible to nutritional modulation; the effects of low vitamin D status during these early life stages cannot completely be rescued by postnatal supplementation. Moreover, vitamin D may impact bone health via gut microbiota‐derived LPS.

Support or Funding Information

Funded by Centrum Foundation Research Innovation Fund, Department of Nutritional Sciences U of T, NSERC, Lawson Family Chair in Microbiome Nutrition Research, Banting and Best Diabetes Centre Novo Nordisk Studentship and Tamarack Graduate Award.
Vitamin D Reduces Hepcidin Concentrations Independent of Inflammatory Cytokines in Healthy Adults: A Pilot, Randomized, Double‐Blind, Placebo‐Controlled Trial of High‐Dose Vitamin D3
Authors: Smith Ellen , Alvarez Jessica , Kearns Malcolm , Hao Li , Sloan John , Konrad Robert , Ziegler Thomas , Zughaier Susu , Tangpricha Vin
Published: November 2023

Objective

Disturbances in iron recycling may result from elevations in pro‐inflammatory cytokines and hepcidin, the major iron‐regulatory hormone. In vitro studies suggest that vitamin D may reduce hepcidin and pro‐inflammatory cytokines, and vitamin D deficiency is associated with increased odds of anemia of inflammation. However, data from clinical trials assessing the vitamin D‐mediated effects on iron recycling in healthy individuals are lacking. Our objectives were to: 1) examine the effect of high‐dose vitamin D on plasma hepcidin, inflammatory cytokine, and ferritin concentrations in healthy adults; and 2) determine whether any observed changes in hepcidin were concomitant with or independent of changes in pro‐inflammatory cytokines.

Methods

This was a pilot, double‐blind, placebo‐controlled trial in healthy adults (N=28) randomized to receive a one‐time oral dose of 250,000 IU of vitamin D3 or placebo. Between‐and within‐group differences in plasma hepcidin, pro‐inflammatory cytokine [interleukin (IL)‐1β, IL‐6, IL‐8, monocyte chemoattractant protein‐1 (MCP‐1)], and ferritin concentrations at baseline and 1 week were determined using two‐sample and paired t‐tests, respectively.

Results

At baseline, plasma 25‐hydroxyvitamin D [25(OH)D], hepcidin, pro‐inflammatory cytokine, and ferritin concentrations did not differ between the two randomized groups; 75% of subjects had plasma 25(OH)D concentrations < 20 ng/mL. After 1 week, plasma hepcidin concentrations had decreased significantly from baseline by 73% in those who received vitamin D3 (geometric mean ratio [GMR]: 0.27 (95% CI: 0.11, 0.62); P=0.005) compared to no significant change with placebo (GMR: 0.73 (95% CI: 0.49, 1.09); P=0.11). Plasma cytokine and ferritin concentrations did not change significantly in either group.

Conclusion

High‐dose vitamin D3 significantly reduced plasma hepcidin concentrations in healthy adults 1 week post‐dosing, while no change was observed in pro‐inflammatory cytokine or ferritin concentrations. These findings suggest that vitamin D may have a role in regulating iron recycling by acting directly on hepcidin, independent of changes in inflammatory markers; however, down‐stream effects on markers of iron status in the absence of inflammatory conditions remain unclear. Longer follow‐up and evaluation of additional markers of iron status may be required.

Support or Funding Information

T32 DK007734 (EMS), K01 DK102851 (JAA), K24 DK096574 (TRZ), UL1 TR000454 (TRZ, VT)
Effects of Calorie Restriction on Microglial Activation in a murine model
Authors: Quijano Uriel , Contreras Irazú , Estrada José A.
Published: November 2023

Calorie restriction has been shown to have beneficial physiological effects in different organisms, delaying the onset of chronic non‐communicable diseases, increasing maximum lifespan, as well as reducing excess body weight and the risk of diseases associated with aging. Recent studies have shown that calorie restriction inhibits the morphological change that is associated with activated microglia, but so far, no data are available to describe if caloric restriction somehow alters the effector functions of microglia.

To determine the effect of calorie restriction on microglia, we analyzed purified mononuclear cells from the brain and spinal cord of 8 weeks old BALB/c mice, divided into three experimental groups: ad libitum, calorie restriction (30% reduction in food intake) and hiper‐energetic (supplemented with sucrose in drinking water), exposed to this diet for 4 weeks. After treatment, brain and spinal cord samples were obtained and microglial cells were purified by Percoll density gradients centrifugation. Purified cells were stained with antibodies for MHC‐I, MHC‐II, CD80 and CD86, as well as cell surface markers CD45 and CD11b and anlyzed by flow cytometry.

Our results show no apparent statistically significant difference in the frequency of MHC+ cells in the experimental groups compared with the control group. The frequency of CD80 and CD86 single positive cells both appear to be increased in the CR group, compared to the control, although the frequency of cells expressing these markers is very low in all groups. We are analyzing the mean fluorescence intensity of all markers to determine if there is a difference in their expression among groups.
In vitro Comparison of Anti‐Inflammatory Activity of Green Tea Catechins
Authors: Fechtner Sabrina , Ahmed Salahuddin
Published: November 2023

Background/Purpose

Green tea contains considerable amounts of epigallocatechin‐3‐gallate (EGCG), epigallocatechin (EGC), and epicatechin (EC). EGCG has already been shown to exhibit anti‐inflammatory activity. Pro‐inflammatory cytokine interleukin‐1β (IL‐1β) produced by monocytes and macrophages plays an important role in rheumatoid arthritis (RA). In this study, we compared the efficacy of different catechins in reducing the inflammatory response to IL‐1β in human RA synovial fibroblasts (RA‐FLS).

Methods

The effect of EGCG, EGC, and EC on RA‐FLS viability was tested using MTT assay. RA‐FLS were pre‐treated with different concentration EGCG, EGC, and EC (5–20 μM) overnight then stimulated with IL‐1β (10 ng/ml) for 24 hours. Condition media was used to quantify IL‐6 and IL‐8 production by ELISA, and MMP‐2 activity by gelatin zymography. The cellular expression of cyclooxygenase‐2 (Cox‐2) and Cox‐1 was evaluated using Western blotting method.

Results

EGCG, EGC, and EC exhibit no toxicity in human RA‐FLS (n=3). Among the tested catechins, EGCG was the most effective catechin at inhibiting IL‐6 and IL‐8 production by 59% and 57%, respectively (n=3; p<0.05). Western blot analysis showed that EGCG selectively inhibits Cox‐2 expression by 86% after IL‐1β stimulation (n=3; p<0.05) and had no significant effect on IL‐1β‐induced Cox‐1 expression. However, EGC not only inhibited IL‐1β‐induced Cox‐2 expression by 58%, it also reduced IL‐1β‐induced Cox‐1 expression by 20% (n=3; p<0.05). Interestingly, our gelatin zymography results showed that EGC was as effective as EGCG in reducing IL‐1β‐induced MMP‐2 activity by 85% in human RA‐FLS (n=3; p<0.05). In all the proposed assays, EC showed no anti‐inflammatory properties.

Conclusion

The results of the study suggest that green tea catechins differentially interfere with IL‐1β signaling pathways to regulate IL‐6 and IL‐8 production, and Cox‐2, Cox‐1, and MMP‐2 expression in human RA‐FLS. Among the catechins tested, EGCG was a more potent anti‐inflammatory compound than EGC and EC in human RA‐FLS. EGCG and EGC display moderate anti‐inflammatory properties while EC displays minimal anti‐inflammatory properties. Together, these results suggest that the gallate moiety is essential to EGCG and EGC inhibiting IL‐1β‐induced inflammatory mediators in RA.

Support or Funding Information

This study was supported by the Robbers Research Award (S.F.), NIH grant AR063104 (S.A.), the Arthritis Foundation Innovative Research Grant (S.A.), the start‐up funds from Washington State University (S.A.).
Cranberry and Recurrent Urinary Tract Infections (UTIs) in Generally Healthy Women: A Systematic Review and Meta‐analysis of Randomized Clinical Trials (RCTs)
Authors: Kern Hua , Liska DeAnn , Chung Mei
Published: November 2023

The aim of the present meta‐analysis was to evaluate the efficacy of cranberry on the recurrence of uncomplicated urinary tract infections (UTIs) among otherwise healthy women. [Registered on International Prospective Register of Systematic Reviews: PROSPERO 2015:CRD42015024439] Two scientists independently conducted the literature search and extracted data, with discrepancies resolved via discussion with a third scientist. Meta‐analysis was performed using RevMan (version 5.3, Cochrane). Six studies (1320 subjects) were included in qualitative and quantitative synthesis. Cranberry reduced the risk of uncomplicated UTI recurrence among healthy women by 18% [Risk Ratio (RR) =0.82; 95% Confidence Interval (CI): 0.68–0.99; p=0.04] (I2=38%). When an outlier study was excluded from the analysis, cranberry further reduced the risk of UTI recurrence by 27%, compared to placebo/control (RR=0.73; 95% CI: 0.60–0.90; p=0.003) (I2=0%). Subgroup analysis indicated cranberry seemed to be more effective in women who were free of UTI at the start of the trial (RR=0.72; 95% CI: 0.55–0.94; p=0.02; n=3 studies) (I2=0%), or over a longer (12 months) follow‐up period (RR=0.61; 95% CI: 0.40–0.91; p=0.02; n=2 studies) (I2=0%), than those who had UTI at the study entry or were followed up for 6 months (p>0.10), respectively. Risk of bias assessment indicated half of the included trials did not provide the randomization information, two had high rates of loss to follow‐up, and two thirds were subject to other biases. Results of this meta‐analysis suggest cranberry can be a potential nutritive option for generally healthy women to prevent the recurrence of uncomplicated recurrent UTIs.

Support or Funding Information

Supported by Ocean Spray Cranberries, Inc.
Quantitative Measurement of Plasma and Urinary Excretion of 3‐Hydroxyisovaleryl Carnitine by UPLC‐MS/MS
Authors: Wang Zhu
Published: November 2023

Objective

An increased plasma concentration of 3‐hydroxyisovaleryl carnitine (3HIA‐carnitine) results from impairment in the leucine catabolic pathway at the conversion of 3‐methyl‐crotonyl‐CoA to 3‐methylglutaconyl‐CoA which is caused by reduced activity of the biotin‐dependent enzyme 3‐methylcrotonyl‐CoA carboxylase. In order to validate plasma 3HIA‐carnitine as a novel biomarker of biotin deficiency rats models, an LC‐MS/MS method for the quantitation of 3HIA‐carnitine in plasma and urinary excretion has been established.

Materials & Methods

Plasma and urinary excretion samples were centrifugated with high speed at 3000×g for 10 minutes, the MCX solid phase extraction column was used to filter and purify the liquid samples, the following procedures were elution, wash‐out, redissolve, detection. The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column (1.7μm, 2.1×50mm), with a mobile phase composed of 0.1% formic acid in water‐methanol with gradient elution. Analyte quantification was performed in the positive electrospray ionization mode and multiple monitoring. The quantification and qualification were performed using the internal standard of 3‐Hydroxyisovaleryl carnitine, then the evaluation for accuracy and precision were followed.

Main Findings

The RSD of six parallel determination for the 3HIA‐carnitine in plasma and urine were 4.8%, 5.5%, respectively. The linear range of 3‐Hydroxyisovaleryl carnitine was 0.5~20ng/ml in the plasma 5~200ng/ml in urine, with both the linear correlation coefficient was above 0.99. Moreover in three standard levels, rate of recovery in plasma and urine were more than 102.5%, 79%. The detection and quantification limits for urine and plasma were 20ng/L, 50ng/L and 10ng/L, 33ng/L, respectively.

Conclusion & Recommondations

The sample preparation method is simple and fast, and the method can be used to analyze 3‐Hydroxyisovaleryl carnitine in plasma and urinary excretion efficiently and sensitively. If confirmed in deeper studies, 3HIA‐carnitine in plasma and urinary is likely to be an important indicator of biotin status in a variety of clinical circumstances especially for the high‐risk population like pregnant women.

Support or Funding Information

Youth fund of national natural science fundation in China (81302428)
Metabolomic Profile of Serum 25‐Hydroxy‐Vitamin D
Authors: Nelson Shakira M , Anic Gabriella M , Panagiotou Orestis A , Mondul Alison , Weinstein Stephanie J , Albanes Demetrius
Published: November 2023

The role of vitamin D in human health has been studied using a variety of approaches. In laboratory studies, the active form of vitamin D, 1,25(OH)2D3 has been shown to play a significant role in reducing autoimmune diseases, such as diabetes. In epidemiological studies, vitamin D has been associated with risk for some cancers but not others. For example, fairly consistent inverse (i.e., protective) associations with circulating 25‐hydroxy‐vitamin D (25(OH)D) have been observed for colorectal cancers, while direct (i.e., adverse) associations have been seen in prostate cancer studies. In order to gain a more comprehensive biological understanding of vitamin D associations with cancers and other health outcomes, we utilized a metabolomics approach to identify metabolites associated with serum 25(OH)D. Over 900 metabolites were measured from fasting blood samples collected at study entry between 1985 and 1988 in the Alpha‐Tocopherol, Beta‐Carotene Cancer Prevention (ATBC) Study, a cohort of Finnish male smokers ages 50–69 years. Baseline serum 25(OH)D was measured in 392 men using radioimmunoassay, chemiluminescence assays, and OCTEIA direct enzyme‐linked immunosorbent assay kits. Metabolites were profiled using ultrahigh performance liquid chromatography/mass spectroscopy and gas chromatography/mass spectroscopy (Metabolon, Inc., N.C.). Metabolite‐vitamin D associations were modeled using linear regression. A statistically significant (p<0.05) change in beta‐estimates per 1 standard deviation (SD) increment was seen for 117 metabolites, the majority of which were lipids, along with 26 amino acids, 10 xenobiotics, seven peptides, six cofactors or vitamins, and four carbohydrate metabolites. Six lipid metabolites were significant at the Bonferroni corrected threshold of p < 0.000053, including 3‐carboxy‐4‐methyl‐5‐propyl‐2‐furanpropanoic acid (CMPF) (beta=0.42/1 SD increment), stearoyl‐arachidonoyl‐glycerophosphoethanolamine (GPPE) (beta =−0.36/1 SD), stearoyl‐linoleoyl‐GPPE (beta = −0.35/1 SD), erucoyl sphingomyelin (beta = 0.32/1 SD), eicosapentaenoate (beta=0.24/1 SD) and docosahexaenoate (beta=0.21/1 SD). The identification of these metabolites may provide insights into the role of vitamin D in human health.

Support or Funding Information

Research support: Intramural Research Program of the National Cancer Institute, NIH.