bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Form of Curcumin Attenuates Performance Decrements Following Muscle Damaging Exercise
Authors: Oliver Jonathan Michael , Caldwell Aaron , Sanders Elizabeth , Mitchell Joel , Rowlands David , Purpura Martin , Jager Ralf
Published: November 2023

It is known that unaccustomed exercise especially that with an eccentric component causes muscle damage and subsequent performance decrements. Attenuating muscle damage may improve performance and allow for greater performance adaptations. Curcumin, a highly pleiotropic molecule, interacts with multiple inflammatory pathways, primarily by inhibiting the IκB Kinase (IKK) signaling complex thereby preventing the activation of nuclear factor‐kappa B (NF‐κB). Curcumin has been reported to blunt the inflammatory response and attenuate performance decrements typically observed following exercise induced muscle damage in mice. However, the efficacy in humans is unknown. Further, poor solubility and rapid metabolism of standardized curcumin may limit potential benefits. CurcuWIN®, a novel curcumin formulation containing 20% curcuminoids (CUR), demonstrates a 45.9 fold greater absorption over standard curcumin. Therefore, the current study sought to examine the effect of differing doses of a novel form of curcumin on performance decrements following downhill running, a known exercise type to result in muscle damage. Sixty‐two moderately trained men and women (21±2 y; 70.0±13.7 kg; 169.3±15.2 cm; 25.6±14.3 BMI) were randomly assigned to ingest 50 mg CUR (in the form of 250 mg CurcuWIN®), 200 mg CUR (in the form of 1,000 mg CurcuWIN®), or placebo (PLA; corn starch) for eight weeks in a double‐blind, randomized, placebo controlled parallel design. At the end of the eight week supplementation period subjects completed a muscle damaging exercise protocol, downhill running. Muscle function, isokinetic peak flexion and extension, was assessed prior to and 1 h, 24 h, 48 h, and 72 h post downhill run. Standardized magnitude based inference was used to define outcomes. As evidence of the exercise bout inducing performance decrements, clear negative contrasts on performance were observed in both the PLA and low dose CUR at 1 h (mean pre post difference range; ±SD range) (extension, −13 to −18; ±5.9 to 7.9 ft‐lbs; flexion, −5 to −6; ±2.2 to 2.6 ft lbs) and 24 h (extension, −10 to −11; ±6 ft‐lbs; flexion, −4 to −6; 3 ft‐lbs) post‐downhill run. High dose CUR attenuated performance decrements, as evidenced by a lack of clear contrasts at 1 h (extension, −8; ±4 ft lbs; flexion −4; ±2 ft‐lbs) and 24 h (extension, −6; ±6 ft‐lbs; flexion, −3; ±3 ft‐lbs). Given the lack of clear contrasts, these data suggest that high dose bioavailable curcumin (200 mg CUR) attenuates performance decrements following downhill running, eccentric loading, which may improve subsequent adaptations to chronic training. Further study is warranted in other exercise types (i.e. resistance training) and chronically.

Support or Funding Information

This study was supported by OmniActive Health Technologies Ltd., Mumbai, India
Benefits of Combinatorial Therapies for Improving Functional Recovery in a Rat Model of Facial Nerve Injury
Authors: Runge Elizabeth M , Martinez Daniel , Drejet Sarah , Brown Brandon , Asante Tony , Welch Haley , Walker Chandler L , Best Amy , Muldoon Jessica , Jones Kathryn J
Published: November 2023

Injury to the facial nerve due to trauma or surgical intervention for head and neck pathologies is common; however, few effective treatments eliciting full functional recovery following facial nerve injuries exist. Previous studies in our laboratory have found that single‐agent therapies, while effective in improving functional recovery, are insufficient in promoting complete functional recovery after nerve injury. Consequently, our objective is to determine the efficacy of combination therapies for treatment of facial nerve injury. We have found that treating the repaired injured nerve with polyethylene glycol (PEG), a sealant that acts to fuse damaged cell membranes, and methylene blue (MB), an antioxidant, improves functional recovery in a rat model of facial nerve transection and suture. Given the ease in application of these therapies, either singularly or in combination, we anticipate that these and future studies will aid translation of such methods for application in human facial nerve injury patients.

Support or Funding Information

Supported by a Roudebush VA IIMR Young Investigator Award (CLW) and IUCRG grant fromm IUSM (KJJ).
Inulin‐Type Fructans Fiber Supplementation Improves Weight Loss When Used with the USDA's MyPlate Recommendations for Weight Management in Obese Women Attempting to Lose Weight
Authors: Salmean Younis
Published: November 2023

The objective was to determine the impact of using weight loss recommendations in free living obese women interested in weight loss, with and without inulin‐type fructans (ITF) fiber over a six‐week period.

Methods

The study was a simple randomized intervention study with twelve women age (22.6 ± 3.6 y), and BMI ≥30 kg/m2. Participants were randomly assigned to either weight management recommendation (WMR) only group (n=6), or WMR+ITF group (n=6). The guided recommendations were a printed guide containing recommendations on how to lose weight based on the United States Department of Agriculture (USDA) MyPlate recommendations. The WMR+ITF group received a six‐week supply of ITF along with the printed guide, while the WMR‐only group received only the guide. Each participant receiving the supplement was instructed to consume one small bag containing 7 g of ITF three times a day before meals. Weight, height and BMI were all measured. In addition, 24 h dietary recall was collected in the beginning (pre‐intervention) and at the end of the six‐week study (post‐intervention).

Results

There were no differences in age, initial weight or BMI between both groups at baseline. Post‐intervention, the WMR+ITF group lost significant weight compared to its pre‐intervention weight (89.2 ± 10.2 vs 87.4 ± 9.9, p<0.05), while the WMR only group did not see a significant change (91.5 ± 12.5 vs 90.9 ± 12.3, p>0.05). Using a 24‐hour recall, the energy intake at pre‐intervention was not statistically different between both groups. Post‐intervention, the WMR+ITF group consumed significantly less energy compared to its pre‐intervention energy intake from 1947 ± 129 to 1742 ± 109, p<0.05, this difference was not see in the WMR only group.

Conclusion

Supplementation with ITF appears to enhance the weight loss achieved with the use of MyPlate recommendations over a short period in obese women attempting to lose weight as evident by greater weight loss in the supplemented group. Decreased energy consumption is likely to explain the weight loss in the supplemented group.

Support or Funding Information

**This work was supported and funded by Kuwait University, Research Project No. (WF02/15).
Alpha‐Lipoic Acid Inhibits IL‐8 Expression by Reducing Oxidative Stress in Ataxia Telangiectasia Fibroblasts
Authors: Yoon Jungyun , Lim Joo Weon , Kim Hyeyoung
Published: November 2023

Ataxia telangiectasia (A‐T) is a neurodegenerative and inherited disease caused by a mutation in the ataxia telangiectasia mutated (ATM) gene. ATM gene is associated with cell cycle arrest, DNA repair, or apoptosis. Reactive oxygen species (ROS) levels were higher in AT cells lacking ATM gene than in normal cells. Previously, we demonstrated that alpha‐lipoic acid inhibits IL‐1 beta‐induced IL‐8 expression by suppressing ROS production in A‐T fibroblasts.

Ataxia telangiectasia (A‐T) fibroblasts were treated with or without alpha‐lipoic acid dissolved in ethanol (final concentration of 10 and 20 microM). ROS levels were determined using DCF‐DA. IL‐8 mRNA expression was determined by quantitative real‐time PCR. Mitochondrial functions such as oxygen consumption (OCR) and ATP production were assessed using Seahorse XF assay. Mitochondria membrane potential (MMP, Δψm) was determined by measuring JC‐1 fluorescent dye using laser‐scanning confocal microscope.

As a result, IL‐1 beta increased ROS levels and induced IL‐8 expression, but decreased mitochondrial functions in A‐T cells. IL‐1 beta‐induced IL‐8 expression and ROS production were inhibited by alpha‐lipoic acid. IL‐1 beta‐induced decrease in mitochondrial function (determined by OCR, ATP production, MMP) was reversed by alpha‐lipoic acid in A‐T cells.

In conclusion, consumption of alpha‐lipoic acid‐containing foods may be beneficial for preventing inflammation of A‐T patients.
Vitamin B12 Status in Women of Reproductive Age, NHANES 2013–2014
Authors: Fothergill Amy , Finkelstein Julia
Published: November 2023

Objective

This analysis was conducted to determine the prevalence of vitamin B12 insufficiency among women of reproductive age who participated in NHANES 2013–2014.

Methods

Women were eligible for inclusion in this analysis if they participated in (NHANES 2013–2014), attended the mobile examination clinic (MEC), had serum vitamin B12 concentrations assessed, and were between 15 and 49 years of age. In the 2013–2014 NHANES sample, women aged 20 to 44 years were eligible for physical examination at the MEC and were classified as pregnant (via urine pregnancy test at MEC; n=61), lactating (i.e., self‐reported through interview; n=39), or non‐pregnant, non‐lactating (n=1,073). Serum vitamin B12 concentrations were assessed via chemiluminescence. Vitamin B12 deficiency and insufficiency were defined as less than 148 pmol/L and less than 221 pmol/L, respectively. Marginal vitamin B12 status was defined as 3148 pmol/L and <221 pmol/L. Linear and logistic regression models were used to examine vitamin B12 concentrations and the prevalence of vitamin B12 deficiency and insufficiency in pregnant and lactating women, compared to non‐pregnant non‐lactating women.

Results

A total of 1,173 women of reproductive age were included in this analysis. A total of 13.3% (n=156) and 2.6% (n=30) were vitamin B12 insufficient and deficient, respectively. Among pregnant women, 21.3% were vitamin B12 insufficient (<221 pmol/L) and 6.6% were classified as vitamin B12 deficient, with an average of 333.9 pmol/L (95% CI: 282.91, 384.89). Among non‐pregnant, non‐lactating women, 13.1% were vitamin B12 insufficient and 2.33% were classified as vitamin B12 deficient, with an average of 419.5 pmol/L (95% CI: 385.27, 453.68). Among lactating women, 7.69% were vitamin B12 insufficient and 2.56% were classified as vitamin B12 deficient, with an average of 497.65 pmol/L (95% CI: 434.5, 560.79). There were no statistically significant differences in vitamin B12 concentrations or the prevalence of vitamin B12 deficiency and insufficiency in pregnant and lactating women, compared to non‐pregnant non‐lactating women.

Conclusions

The prevalence of vitamin B12 insufficiency was relatively high among pregnant and non‐pregnant women of reproductive age in the United States.
Dietary Habits among Adolescent Girls 9–13 Years of Age that have Accessed Nutritional Information in Last 12 Months in the Upper Manya Krobo District, Ghana
Authors: Ghadirian Mona Ziba , Marquis Grace , Clark Shelley , Aryeetey Richmond , Obeng Dinah
Published: November 2023

A desire to acquire health information is present among Ghanaian youth. About half (53%) of adolescent Internet users, 15–18 years of age residing in Accra, Ghana, have searched for health information online. Access to adequate nutritional information may positively influence dietary habits among adolescent girls. This analysis examined the association between the acquisition of nutritional information and dietary habits among adolescent girls. We carried out a cross‐sectional survey among adolescent girls 9–13 years of age (n=1420) living in 3 subdistricts of the rural Upper Manya Krobo district in the Eastern Region of Ghana in 2014. Data were recorded concerning the access to nutritional information during the past 12 months as well as dietary intake during the previous week. Ten percent of adolescent female participants reported receiving nutritional information during the previous 12 months. Among those who did receive nutritional information, the majority of participants (62%) received it in school, followed by access from home (21%), local community activities (9%), local clinic (6%), and other sources (1%). Receiving nutritional information was significantly associated with consumption of organ meats (p < 0.0001) whereas there was no significant association with diet diversity (p=0.14), intake of animal source foods (p=0.35), or vitamin A‐rich food products (p=0.87). Research shows that nutritional information is not yet accessible to the majority of adolescent girls in this district. Further research is needed to identify best nutrition education practices that increase the accessibility and quality of health messages tailored to adolescent girls. These data are part of Clinical Trials # NCT01985243.

Support or Funding Information

Global Affairs Canada, Government of Canada.
Inadequate Sleep Quality On‐and Off‐duty Among Central Texas Male Firefighters
Authors: Hernandez Ladia M. , Loza Colleen , Dong Kane , Forman Michele R
Published: November 2023

According to the Bureau of Labor Statistics, firefighters have one of the highest rates of illnesses and injuries of all occupations. Occupational stress is associated with diseases and health issues seen in emergency responders. It has been established that poor sleep quality, sleep disruption and inadequate sleep among the most important causes of occupational‐stress. Chronic sleep loss in addition to long work hours are associated with adverse health consequences. Poor sleep quality has been linked to increased prevalence of obesity, Type 2 diabetes, anxiety, depression, and cardiovascular disease (CVD). The Exploring Health and Lifestyle in Firefighters Study (EHLF) seeks to identify risk factors and improve disease outcomes in firefighters. The present study aims to evaluate overall sleep quality of Central Texas firefighters – specifically, the differences between the on‐duty and off‐duty days as well as seasonal effects. Demographics, health history, anthropometrics, clinical data, and sleep quality data were collected for 79 male firefighters. The validated Pittsburg Sleep Quality Index (PSQI) was administered for Department A over summer of 2014 and winter of 2014–2015 and for Department B over summer of 2015 and winter of 2016. PSQI questionnaires were administered during both seasons to independently evaluate on‐duty sleep quality and off‐duty sleep quality. PSQI total scores indicated that 76% of firefighters reported poor sleep quality during the summer on‐duty period and 43% reported poor sleep quality during off‐duty days. During the winter on‐duty period, 77% reported poor sleep quality whereas for the winter‐off duty period, 44% reported poor sleep quality. These results, for the first time, highlight the need to address the sleep quality of firefighters while on‐ and off‐duty. The majority of firefighters reported poor sleep quality while on‐duty, which has acute and chronic health implications. Future research should focus on identifying methods to improve on‐duty sleep quality in firefighters.
Association of dietary sugar intake with bone loss, non‐traumatic fracture risk, and all‐cause mortality in Chinese elderly: finding from an 11‐year longitudinal study of Mr and Ms OS Hong Kong
Authors: Liu Zhaomin
Published: November 2023

Background

The association of dietary sugar intake and skeletal health remains uncertain, especially among elderly. Our objective was to investigate sugar consumption in Chinese elderly and the temporal association of sugar intake with bone health and mortality.

Methods

Analyses were conducted based on an 11‐year longitudinal study (Mr and Ms OS Hong Kong). A total 4000 Chinese men and women aged 65 and older were recruited from the local community from 2001 to 2003. The study consisted of a 4‐year follow‐up (n=3416) on the changes in bone mineral density (BMD) and an 11‐year follow‐up for the incidence of non‐traumatic fracture and all‐cause mortality (n=3344). Free and added dietary sugar intakes were assessed at baseline using a 329‐item validated Food Frequency Questionnaire. BMD was examined at baseline and 4‐year follow‐up by dual‐energy X‐ray absorptiometry at total hip, spine, femoral neck and whole body. Data on the incidence of non‐traumatic fractures and all‐cause mortality were collected. General linear model, logistic, and Cox proportional hazards models were used to test the associations of sugar intakes with bone health and all‐cause mortality.

Results

The average free sugar intake among the participants was 20.0±19.8 g/d (4.16±3.57% calories) with only 5.4% exceeding the WHO recommendation (<10%). During a median follow‐up of 11.1 years (34,483 person years), we documented 433 non‐traumatic fractures and 769 deaths. No significantly association was observed between total free and added sugar intakes with BMD changes, as well as the incidence of non‐traumatic fracture. However, high sugar intakes were significantly associated with low risk of all‐cause mortality in elderly women with hazard ratios of 0.625 (95% CI: 0.428–0.915, p for trend: 0.033) for free sugar and 0.578 (95% CI: 0.386–0.867, p for trend: 0.004) for added sugar in the highest quintile compared with that in the lowest quintile.

Conclusions

The amount of sugar consumed by Chinese elderly pose no risk to bone loss and fracture. High sugar intakes were even associated with low all‐cause mortality in elderly women.

Support or Funding Information

The study was supported by the National Institutes of Health R01 grant AR049439–01A1 and the Research Grants Council Earmarked Grant CUHK4101/02 M. The funders played no role in the study design, data collection and analysis, interpretation of the data, as well as in writing the manuscript.
Are Meals and Snacks Consumed by Preschool‐aged Children in Family Child Care Homes Meeting the Dietary Guidelines?
Authors: Tsai Maggie You Ming , Mena Noereem Z , Gans Kim M , Risica Patricia M , Lofgren Ingrid E , Tovar Alison
Published: November 2023

Although family child care homes (FCCH) are the second most utilized form of non‐relative child care in the US, little is known about what children eat in this setting. The objective of this study was to assess the quality of meals and snacks consumed by children aged 2–5 in FCCH compared to the 2015 Dietary Guidelines for Americans (DGA 2015). This study used baseline data from an ongoing cluster randomized controlled intervention trial in FCCH. Preschool‐aged children (n = 104; 2–5 years old) enrolled in 34 FCCH, were observed during two meals and a snack each day for two days, following a standardized protocol. Foods and beverages consumed were analyzed in the Nutrition Data System for Research 2015. Mean portion sizes consumed of each food group were compared to 2/3 of the daily intake recommendations established by the DGA 2015 for children 1–3 years old (1000 kcal) as recommendations are not standard for children 2–5 years old, proportional to the daily nutrient requirements for time spent in childcare. One‐sample t‐tests were run to test for differences between the mean amounts of food groups consumed compared to the national guidelines. Mean intakes of children did not meet the recommended 0.67 cup of vegetables (0.27 ± 0.21 cup, 40% of recommendation), 1.0 ounce of whole grains (0.37 ± 0.37 ounce, 37% of recommendation), and 14 grams of fiber (5.60 ± 2.14 gram, 40% of recommendation). However, children were close to meeting the recommended 0.67 cup of fruit (0.64 ± 0.46 cup, 97% of recommendation). Generally, children in this FCCH study did not meet national recommendations for vegetables, whole grain, and fiber. Future research should continue to develop and evaluate strategies to increase fruit, vegetable and whole grain intake in early child care settings.

Support or Funding Information

This project was supported by the National Heart, Lung, And Blood Institute of the National Institutes of Health under Award Number R01HL123016
Genetics of acute kidney injury markers in Hispanic children: the Viva La Familia Study.
Authors: Vazquez‐Vidal Itzel , Chittoor Geetha , Mass Baba , Haack Karin , Laston Sandra , Mehta Nitesh , Comuzzie Anthony G , Cole Shelley , Butte Nancy , Voruganti V. Saroja
Published: November 2023

Studies have demonstrated that chronic kidney disease in adults may develop from acute kidney injury during childhood. Thus, increases in acute kidney injury (AKI) markers that are indicative of sharp decline in kidney function are a cause of concern for later kidney disease. AKI markers are also under considerable genetic influence. Our aim in this study was to identify genetic determinants of urinary AKI markers: neutrophil gelatinase‐associated lipocalin (NGAL), cystatin C (CysC), beta‐2‐microglobulin (β2M), osteopontin (OST) and trefoil‐factor 3 (TFF3) in Hispanic children of the Viva La Family Study. Urinary concentrations of NGAL, CysC, β2M, OST and TFF3 were measured in 837 children. We conducted a genome‐wide association analysis of AKI markers after accounting for family kinships. All methods were implemented in SOLAR. Empirical thresholds for genome‐wide significant and suggestive associations were defined as p <1 × 10−7, and p <1 × 10−6, respectively. Significant heritabilities were found for all AKI markers (range 35 – 68%), with CysC showing the highest heritability (h2= 0.68, p= 1.9×10−15). The GWAS analysis showed strong association of rs7555367 of phospholipid phosphatase related 5 (PLPPR5) gene with OST (p = 3.5 × 10−8, MAF = 0.09, β coefficient = −0.58 (0.10)) and suggestive association with TFF3 and β2M (p < 2.7 × 10−6, MAF = 0.09, β coefficient = −0.48 and −0.55 (0.11)). We also found suggestive association between OST and 8 SNPs (rs10169879, rs10210408, rs16856600, rs2075252, rs2284675, rs2389589, rs3821125, rs4668123) in lipoprotein‐related protein 2 (LRP2)/megalin gene (p < 3 × 10−7) and between NGAL and rs12947457, rs7216231 and rs4968239 of N‐ethylmaleimide sensitive factor, vesicle fusing ATPase (NSF) (p < 3.4 × 10−7). These genes have key roles in cell signaling and signal transduction. In summary, we observed significant genetic influence on the variation in AKI markers and identified a potential role for PLPPR5, LRP2 and NSF in AKI in children.

Support or Funding Information

The National Institutes of Health (NIH) [R01DK080457, R01MH59490, P30ES010126 and R01DK092238] and the USDA/ARS [Cooperative Agreement 6250‐51000‐053].
Transgenerational Effects of Branched Chain Amino Acids Supplement Combined with High Fat Diet in Male Mice
Authors: Han Jihee , Lee Yeon Kyu , Park Joo Hyun , Park Yoon Jung
Published: November 2023

Consumption of branched chain amino acid (BCAA) as a supplement has dramatically increased among athletes and even the general public in the last decade. Although possible positive effects in energy balance, glucose homeostasis, and muscle endurance have been, they are still controversial depending on various subjects with different age, exercise, and diet intake. Moreover, whether effects of BCAA supplement along with metabolic stress is unknown. In this study, we investigated whether BCAA supplement influence the response to metabolic stress in male mouse fed with high fat diet (HF) and whether the effects influence the next generation. Male mice were divided into three groups each of whom consumed control diet (CD), HF or HF with 2% BCAA supplemented water (HF/BCAA). The males at 15 weeks of age were mated with CD‐fed female mice to generate the offspring. The offspring of all groups was fed the same diets, CD for the first 8 weeks and then HF for later. In the sire, HF feeding regardless of BCAA supplement increased body weight, organ weights, especially adipose tissues, and glucose levels during glucose tolerance test (GTT) and insulin tolerance test (ITT), compared to CD group. BCAA supplement with HF did not show significant differences in the measurements above compared to HF group, except a marginal reduction of body weight sporadically. In male offspring, they have no difference in body weight and glucose level during GTT at 6 weeks of age for consuming CD. After changing to HF, the male offspring of the sire fed HF shown increased body weight, organ weights, and glucose level in GTT at 13 weeks of age compared to the offspring of the sire consuming CD. Intriguingly, the offspring of the sire consuming HF/BCAA showed alleviated glucose impairment and marginally reduced body weight compared to the offspring of the sire consuming HF. In addition, Metabolic alteration in the sire altered gene expression pattern related with glucose metabolism, like as glucokinase in the male offspring. In female offspring, there are no robust change in body weight and metabolism health throughout the whole period, except the initial difference in glucose metabolism during GTT. In conclusion, these data reveal that metabolic stress induced by HF influences not only the subjects, but also the next generation in a sex‐specific manner. Moreover, BCAA supplement attenuates negative transgenerational effect of high fat diet by decreasing body weight and glucose level in GTT like as control diet in male offspring in a sex specific manner although it has no beneficial effects in body weight and glucose metabolism in the mice with diet‐induced obesity.
Red Pepper Seed Extract Suppressed Gluconeogenesis in AML12 and Stimulated Glucose Uptake in C2C12
Authors: Kim Haeun , Go Gwang‐woong
Published: November 2023

Red pepper seed, a by‐product of red pepper, has been reported to have anti‐oxidant and anti‐obesity activity. However, its role in diabetes has not yet been investigated. Glucose homeostasis is maintained by insulin, which suppresses glucose production in liver and enhances glucose uptake in peripheral tissues. In this study, we examined the effect of red pepper seed (RPS) extract on glucose production in AML12 hepatocytes and glucose uptake in C2C12 myoblast. Its underlying mechanism was also investigated. RPS reduced the glucose production in a dose‐dependent manner in AML12; 200 μg/mL of RPS lowered glucose production by 29.25% compared to control (p<0.01). The level of glucose 6 phosphatase and phosphoenolpyruvate carboxykinase, critical enzymes of hepatic gluconeogenesis, were decreased by 200 μg/mL of RPS compared to control (p<0.001 and p<0.01, respectively). In addition, 200 μg/mL of RPS increased the glucose uptake by 46.99% compared to control in differentiated C2C12 (p<0.05). Consequently, insulin‐dependent pathway was analyzed; the activity of IRS1‐pY612, IRS1, and PI3K‐pY199 were significantly activated by RPS (p<0.05). In conclusion, RPS would improve glucose homeostasis via reducing hepatic gluconeogenesis and increasing peripheral glucose uptake. These results suggest that RPS is a compelling nutraceutical for anti‐diabetes.
Effect of rapid or slow weight reduction on the body composition and body energy loss in rats
Authors: Maeda Megumi , Kondo Emi , Inoue Haruna , Okamura Koji
Published: November 2023

Rapid weight loss has been shown to result in greater reductions in the lean body mass (LBM) than slow weight loss, suggesting that less body energy is lost with rapid weight reduction due to the lower energy content of lean tissue compared with fat tissue. The aim of this study was to compare the changes in the LBM, fat mass (FM), and body energy loss between rapid and slow weight reduction. Male Sprague‐Dawley rats were divided into a rapid weight reduction group (Rapid, n = 5), which was fasted for 3 days, or a slow weight reduction group (Slow, n = 5), which had its food intake restricted for 13–21 days to achieve a final body weight comparable to that in the Rapid group. All rats were maintained individually in a metabolic chamber to measure energy expenditure. The initial body weight was 688.0 g (standard deviation [SD] 78.1) in the Rapid group and 693.6 g (SD 34.5) in the Slow group. The body weight reduction and total energy expenditure during the experiment were 63.7 g (SD 12.0) and 183.0 kcal (SD 12.8) in the Rapid group and 63.3 g (SD 11.3) and 1241.2 kcal (SD 245.6) in the Slow group, respectively. The metabolizable energy (absorbed energy minus expended energy and excreted energy into urine) during the experiment in the Slow group was 235.3 kcal (SD 90.7). Consequently, the calculated energy loss for body weight loss by 1 kg did not differ significantly between the Rapid group (2943.3 kcal [SD 405.2]) and the Slow group (3628.9 kcal [SD 1072.6]). The estimated decrease in the LBM and FM based on the energy density of lean tissue (1.1 kcal/g) and fat tissue (7.6 kcal/g) was 46.4 g (SD 12.3) and 17.4 g (SD 1.0) in the Rapid group and 37.8 g (SD 10.1) and 25.5 g (SD 12.6) in the Slow group, respectively, with no significant differences observed between the groups. In conclusion, neither the decrease in the LBM or FM nor in the body energy loss due to body weight loss differed significantly between the rats experiencing rapid and slow weight reduction.
Casein before bed is as effective as receiving earlier in the day in increasing muscle mass and strength in response to resistance training
Authors: Joy Jordan , Vogel Roxanne , DiMarco Nancy , Broughton Ken , Wildman Rob
Published: November 2023

Amino acids from food directly initiate muscle protein synthesis and recent studies support that this process is not altered during sleep. This opens the concept of ingesting protein before bed as part of a greater strategy to maximize effects of resistance training with protein throughout the 24‐hour period. The purpose of this study was to investigate the response of supplemental protein (casein) provided earlier in the day versus before bed when resistance training occurred before 4pm. Young males (19–24 yoa) engaged in supervised resistance training with nutrition guidance and either received casein protein (35g) earlier in the day or before bed for 10 weeks in an isocaloric and nitrogenous design. Data were analyzed using repeated measures ANOVA with Bonferroni post‐hoc. Both groups demonstrated increases in lean soft tissue and strength (p < 0.05) with no difference between groups. No changes were observed for measures of body fat. Providing protein before sleep is an effective time to incorporate protein as part of a 24‐hour strategy, even if the training occurs earlier in the day.

Support or Funding Information

Supported by Dymatize Nutrition
Rifampicin and Isoniazid Induced Liver Injury via the Pregnane X Receptor
Authors: Brewer Christopher Trent
Published: November 2023

Background

Drug‐induced liver injury (DILI) is a significant cause of anti‐tubercular therapy failure. Rifampicin and isoniazid co‐treatment have been shown to lead to cholestatic liver injury associated with the accumulation of protoporphyrin IX (PPIX, a heme precursor) in a human pregnane X receptor (hPXR) dependent manner in mice. Therefore, the activation of hPXR by rifampicin can lead to DILI via PPIX accumulation, thus antagonism of hPXR and modulation of PPIX transport and degradation can prevent the clinical consequences of DILI.

Objective

Using an hPXR mouse model and primary human hepatocytes we will induce liver toxicity and evaluate the potential of a novel small molecule inhibitor of hPXR developed by our laboratory to prevent liver injury and Mlkl activation associated with rifampicin and isoniazid treatment.

Methods

PPIX accumulation was assessed by fluorescence and LC/MS/MS in human cell lines and humanized transgenic mice. hPXR target gene expression was evaluated using Western Blot and qRT‐PCR analyses in human primary cells and humanized mice. Liver injury was evaluated by analyzing mouse liver enzyme activity in sera and via histopathologic examination.

Results

ALAS1/Alas1 (rate‐limiting enzyme of heme synthesis) is increased in response to isoniazid in both hPXR transgenic mice and PXR knockout mice. FECH/Fech (heme synthesis enzyme) is reduced in response to isoniazid in both mouse strains. PPIX is increased in the liver of hPXR mice treated with both rifampicin and isoniazid. Rifampicin and isoniazid co‐treatment activate necroptosis markers in hPXR mice.

Conclusions

Antagonism of hPXR with a small molecule may be a viable therapeutic strategy to reduce liver injury.

Support or Funding Information

Funding provided by ALSAC and NIGMS (GM 086415 & 110034)
Regulation of extracellular matrix gene expression and biosynthetic pathways in pulmonary fibroblasts by HSF1
Authors: Jones Mikayla , Wales Christina TK , Gristock Rachel A , Jacobs Aaron T
Published: November 2023

Idiopathic pulmonary fibrosis (IPF) is a chronic and rapidly progressing lung disease that is characterized by extensive tissue remodeling, impaired gas exchange, and a poor prognosis. The pathogenesis of IPF involves epithelial cell death, leukocyte infiltration and cytokine production, enhanced deposition of collagen and remodeling of the extracellular matrix (ECM). One key event in the progression of IPF is the activation of fibroblasts that is mediated by TGF‐beta, which leads to tissue remodeling. We have investigated a possible role for the transcription factor, heat shock factor 1 (HSF1) in the expression of pro‐fibrotic genes in TGF‐beta treated fibroblasts. siRNA was used to silence the expression of HSF1 in pulmonary fibroblast cell lines, MRC‐5 and IMR‐90. Cells were treated with TGF‐beta for 0, 8, and 24 h, after which RNA and protein samples were collected. RNA samples were analyzed by real‐time PCR to assess differences in the expression of several target genes with known roles in the production and remodeling of ECM, and results were verified by Western blot. Silencing HSF1 altered the expression of several ECM‐related genes in both cell lines, indicting role for HSF1 in the control of ECM biogenesis and remodeling by TGF‐beta stimulated fibroblasts, and a possible significance for HSF1 activity in the etiology of IPF. In addition to components of fibrillar collagen (COL1A1, COL1A2), the expression of several genes that function in the biogenesis of collagen fibers also were impaired by HSF1 silencing, including proteins with proline hydroxylase (P4HA2), lysyl hydroxylase (PLOD1), prolyl isomerase (FKBP4); and chaperone activities (SERPINE1, SERPINH1). In contrast to its effects on fibrillar collagen biogenesis, HSF1 silencing enhanced the expression of genes involved in elastic fibers production, including: elastin (ELN) microfibrillar‐associated protein 4 (MFAP4); and fibulin 5 (FBLN5). This observation is consistent with the finding by others that the ratio of collagen and elastin fibers is altered in IPF, further indicating a unique role for HSF1 in controlling the composition of the ECM in this disease.

Support or Funding Information

Funding provided by the Leahi Fund of the Hawaii Community Foundation
Lack of effect of the direct renin inhibitor aliskiren on cardiac mast cells and macrophages in an acute model of fat embolism in rats
Authors: Fotouhi Ariana , Arif Dauod , Siddiqi Ahsan , Ponnapureddy Rakesh , VanDillen Michael , Tappeta Kathyayiri , Khan Sultan , Hamidpour Soheila , Poisner Alan , Molteni Agostino
Published: November 2023

Introduction

Fat embolism (FE) induced by triolein injection causes both acute and chronic lung injury in rats and the histopathological damage is not limited to the lungs but also involves the heart (1). The renin‐angiotensin system (RAS) plays a role in the pathogenesis of such damage. Captopril and losartan treatment prevented such damage both in lungs and heart at 10 weeks after triolein injection (2). More recently we reported that the renin inhibitor aliskiren induced the same protective effect in the lungs (3) and reduced the number of mast cells 48 hours after triolein (4). The hearts of the same triolein treated rats however had a modest but statistically significant increase of alpha‐smooth muscle actin (SMA) (p<0.05) versus controls but no fibrosis or vasculitis and no changes by aliskiren in the acute experiment. We measured the mast cells and the macrophages in the heart to delineate their potential role in this early cardiac damage.

Methods

Twenty‐two rats were divided into controls (n=4, Group A) and three FE groups (n=6, groups B,C &D). FE induction was done with triolein 0.2 ml i.v at 0 hours, and an hour later animals received 0.2 ml i.p saline (Groups A&B), aliskiren at 50 mg/kg (group C) or at 100 mg/kg (Group D). Rats were euthanized at 48 hours and hearts were fixed and stained with H&E for lumen patency measurements, Masson's Trichrome (MT) for collagen, and SMA for myofibroblasts. Mast cells were stained with CD 117 and macrophages with CD 68.

Results

As reported above, the only damage in the heart of rats treated with triolein and saline was an increase in SMA staining around the branches of the coronary arteries compared to the control group. No other significant difference was observed for all the other parameters. Mast cells and macrophages did not show any statistical difference among the four groups. The macrophages did not show the two types of cells that we have observed in the lungs, one with large cytoplasm with small vacuoles and the second with small compact cytoplasm (5).

Conclusion

Despite the significant cellular response in the lungs at 48 hours after triolein, there were no alterations in mast cells or macrophages in the heart after triolein or aliskiren treatments. In contrast to the lungs, where we have observed two types of macrophages, only one type was present in the heart. The lack of change in mast cells and macrophages in the heart and no fibrosis with only a modest change in SMA at this early time suggests that their involvement may be limited to later responses to fat embolism.

Support or Funding Information

Supported by Mary Katherine Geldmacher Research Foundation, St Louis MO.
Preliminary data on the effects of high fat diet, olive oil and vitamin D on fatty liver and muscle in an experimental model
Authors: Trovato Francesca M , Musumeci Giuseppe , Szychlinska Marta , Nsir Houda , Magro Gaetano , Salvatorelli Lucia , Castrogiovanni Paola
Published: November 2023

Background and Aims

Vitamin D (VitD) is emerging for putative immunomodulatory and antifibrotic effect, and its deficiency is associated with NAFLD and sarcopenia. Olive oil is deemed as a factor of the effect of the Mediterranean diet nutritional profile. We investigated the role of VitD supplementation or deprivation in sedentary rats fed standard diet, high fat diet with animal fat or extra‐virgin olive oil (EVOO), studying the development of fatty liver disease and muscle tissue change.

Methods

In this study 28 male rats, 7–9 weeks of age, 200 g of body weight, on a 12‐hour light/dark cycle and ad libitum access to assigned diet and water, were randomly divided in 7 groups and fed for 10 weeks with 7 different dietary profiles [Regular diet (1); Regular diet + VitD (2), Regular diet VitD deprived (3); High fat/butter + VitD (4) and VitD deprived (5); High fat/EVOO plus VItD (6) and VitD deprived (7)]. Microscopic evaluation and histochemical study of liver and muscle was performed. The NAFLD Activity Score (NAS), ranging from 0 to 8, i.e. for steatosis (0–3), lobular inflammation (0–3), and ballooning (0–2), and the automated muscle fibers image analysis were used.

Results

No difference was found among the 7 groups for final body weight gain, although rats fed a standard diet VitD deprived (3) had the final lower weight; the high fat/EVOO plus VitD rats (5) were heavier vs. controls (p 0,07). Rats on high fat/butter diet, both VitD enriched (4) or deprived (5), and high fat/EVOO VitD deprived (7) developed at last significantly greater NAS. Differently, rats on regular diet (1), notwithstanding with the adjunct (2) or deprivation of VitD (3), and rats fed with high fat/EVOO VitD enriched diet (6) did not show evidence of NAS increase. This last group (high fat/EVOO plus VitD) showed slight lobular inflammation without signs of steatosis, and their muscle fibers were hypertrophic respect to control and to other groups. In contrast, in group 5 (high fat/butter VitD deprived) the fibers were hypotrophic.

Conclusions

VitD seems to be protective against steatosis in rats fed with EVOO‐based high fat diet, but not with butter‐based high fat diet. Moreover, VitD deficiency is associated with muscle fibers hypotrophy, effect counteracted by olive oil‐based diet. Extra‐virgin olive oil and Vitamin D may share a protective role on liver and muscle in sedentary rats against the induction of fatty liver and sarcopenia.
Parathyroid Hormone‐Related Protein Stimulates Head and Neck Cancer Cell Growth via RUNX2 Regulation and Serves as a Poor Prognostic Marker in Patients
Authors: Chang Wei‐Min , Hsiao Michael
Published: November 2023

Parathyroid Hormone‐related Protein (PTHrP) is a paracrine/autocrine ligand and up‐regulated in head and neck squamous cell carcinoma (HNSCC) patients. However, the regulatory mechanism and cellular function in HNSCC cancer remains obscure. We investigated the clinical significance of PTHrP in HNSCC patients, and verified the regulation mechanism in RUNX2/PTHrP/Calcium axis, which is stimulated HNSCC cell growth in auto‐feedback loop. PTHrP acts as a poor prognostic marker in HNSCC patients. PTHrP expression lead to increasing the cell proliferation potential through a paracrine/autocrine mechanism and elevating blood calcium level in immunodeficiency mice. PTHrP is found to be co‐expressed with RUNX2 in public HNSCC microarray cohorts. PTHrP is regulated by RUNX2 and also acting as key calcium regulator in Physiology condition. We found elevations of calcium concentration also increased the RUNX2 expression in HNSCC cells. PTHrP, calcium, and RUNX2 form a positive feedback loop in HNSCC cells. Moreover, ectopic RUNX2 expression also increased PTHrP expression and promoted proliferation potential through autocrine PTHrP expression. Using cDNA microarray analysis, we found PTHrP stimulated expression of cell cycle regulators, namelyCDC25A, CCNA2, and CCNE2 in HNSCC cells, and these genes are also up‐regulated in HNSCC patients. In summary, our results reveal that RUNX2‐PTHrP/Calcium axis contributes to HNSCC tumor growth and PTHrP expression is a poor prognosis marker in HNSCC patients.
Identification of CtBP Targets in Breast Tumor Sections by Proximity Ligation Assay
Authors: Park Samson , Jones Alana , Yi Dae Ik , Liang Genqing , Kabbout Mohamed , Flanders Kathy , Wakefield Lalage M. , Byun Jung S.
Published: November 2023

CtBP (Carboxyl‐terminal binding protein) is a dimeric nuclear protein and an epigenetic regulator that is activated in the presence of NADH to recruit and target a variety of histone modifying complexes and transcriptional regulators to chromatin, thus providing a defined mechanism through which carbohydrate metabolism can drive epigenetic regulatory events. We have recently shown that CtBP drives epithelial‐mesenchymal transition (EMT), genome instability, and acquisition of tumor‐initiating stem cell‐like pathways in breast cancer. Recently, we observed that elevated CtBP protein expression is correlated with poor survival of breast cancer patients in two independent cohorts. This finding led us to study further what complexes are associated with CtBP in breast cancer patient samples and whether such complexes could be linked to pathways that influence clinical outcomes. Immunoprecipitation assay of CtBP followed by mass spectrometry revealed CtBP's interactions with BRG1 (SMARCA4) and ZNF217 in the MCF‐7 breast cancer cell line. ZNF217 is a Zinc Finger DNA binding Transcription factor located on chromosome 20q13. ZNF217, amplified in 8–30% of breast cancers, is capable of transforming primary mammary epithelial cells, has been seen in hyperplastic mammary lesions, and plays a major role in TGF‐beta signaling pathways. BRG1 is a subunit of the SWI/SNF chromatin remodeling complex. The SWI/SNF is an ATP‐dependent chromatin remodeling complex that plays a role in a variety of cancers. Most notably, its subunits are mutated in 20% of cancers, with lesions that either disrupt tumor suppressor activity or confer oncogenic properties on the SWI/SNF complex.

To assess whether these complexes form in patient tumor samples, we employed the Proximity Ligation Assay (PLA) technology. PLA enables detection of protein:protein interactions with sensitivity at the level of single molecule resolution in tissue. We performed PLA on formalin fixed and paraffin embedded (FFPE) tumor samples from breast cancer patients and could demonstrate significant levels of CtBP1:BRG1, CtBP2:BRG1 and ZNF217:CtBP1 interactions. Recent studies from our lab and those of others suggest a cooperative interaction between CtBP and BRG1 in the induction of drug resistance genes. To assess functional relationship between CtBP and BRG1 in patient samples, we measured the transcript abundance of the multiple drug resistance gene MDR1 (ABCB1) in patient samples stratified by BRG1 and CtBP expression. The findings show a significant correlation between MDR1 transcript abundance in patients with both high levels of BRG1 and CtBP (P‐value = 0.021). This data suggest future applications of BRG1/CtBP PLA as a possible screen for the activation of drug resistance pathways in breast cancer patients. Finally, we provide evidence that adopting this assay to tissue microarray (TMA) format is highly feasible.
Implication of Aberrant Protein Expression in the Etiology of Pancreatic Cancer
Authors: Nyaga Simon G , Akobundu Blessing , Giwa‐Otusajo Jamiu , Hazel Kaisha
Published: November 2023

Pancreatic cancer is a leading cause of cancer mortality in the U.S. American Cancer Society estimates that there will be 53,670 new cases and 43,090 pancreatic cancer death in 2017 in the U.S. Although many risk factors are known, the precise mechanisms involved in the development/progression of the malignant phenotype of this devastating disease are poorly understood. The interaction of the risk factors with cellular DNA may result in abnormal protein changes that could affect the functions of essential biological pathways in cells. The goal of this study is to understand the molecular mechanisms involved in the etiology of pancreatic cancer. To address our goal, we focused on identification of differentially expressed proteins in a pancreatic cancer cell line, BxPC‐3, relative to a nonmalignant pancreatic cell line, HPDE‐6. The cell lines were cultured and cell‐free extracts prepared followed by protein separation by SDS polyacrylamide gel electrophoresis (SDS‐PAGE) and staining using Coomassie blue. Specific candidate proteins were identified by liquid chromatography in tandem with mass spectrometry (LC‐MS/MS). Several proteins were shown to be differentially expressed including; MYH10, CEACAM6, and IGFBP‐5 which were confirmed to be highly over‐expressed (approximately 5×, 10× and 5× respectively) in BxPC‐3 cell line compared to HPDE‐6 cell line using Western Blot analysis. Interestingly, these proteins are associated with biological processes such as cell proliferation, differentiation, apoptosis, cell adhesion, and metastasis: processes commonly altered in many cancers. This aberrant protein expression could be associated with abnormal protein functions and interactions that may be critical in the initiation/progression of pancreatic cancer. Hence, these findings implicate IGFBP‐5, CEACAM6, and MYH10 in the etiology of pancreatic cancer. Further biochemical characterization may yield biomarkers useful in effective therapeutic intervention of pancreatic cancer.

Support or Funding Information

Department of Biology, Morgan State University