bluuebunny/crossref_metadata_2025 huggingface dataset:
Decreased Muscle Health In Dietary Antioxidant Deficient Aged Mice
Authors: van Dijk Miriam , Dijk Francina , van Norren Klaske , Jaspers Richard , van Helvoort Ardy , Verlaan Sjors , Luiking Yvette
Published: November 2023

Background and aim

Inadequacies of micronutrients with antioxidant‐properties are common among older adults and have been associated with higher risk of frailty, adverse functional outcome and impaired muscle health. However, a causal relationship is less well known. The aim was to determine in old mice the impact of reduced dietary intake of vitamins A/E/B6/B12/Folate, selenium and zinc on muscle mass, oxidative capacity, strength and physical activity (PA) over time.

Methods

21‐months‐old mice were fed either AIN‐93M or a diet low in micronutrients with antioxidant‐properties (=LOWOX: 50% of mouse recommended daily intake of vitamin A/E/B6/B12/folate/selenium/zinc) for 4 months. Muscle mass, grip strength, PA, and general oxidative status (liver malondialdehyde (MDA)) were assessed and m. tibialis anterior (TA) was used to determine ex vivo muscle protein synthesis (MPS via SUnSET method) and breakdown (MPB via 3‐methylhistine release). Moreover, muscle fatigue was measured on m. extensor digitorum longus (EDL) during an ex‐vivo moderate exercise‐protocol. Effects on oxidative capacity (succinate dehydrogenase (SDH) activity), muscle fiber type, number and cross‐sectional area (FCSA) were assessed on m.plantaris (PL) using histochemistry.

Results

From 2 months on the diet, bodyweight of LOWOX‐mice was lower compared to control‐mice (P<0.0001) without difference in food intake, and mainly due to lower fat mass (P<0.0001). After 4 months, oxidative status of LOWOX‐mice was lower, demonstrated by decreased vitamin E plasma levels (P<0.05) and increased liver MDA levels (P=0.018). PA was significantly lower in LOWOX‐mice (P<0.001). Muscle mass was not affected, although PL‐FCSA was decreased (~16%; P=0.028) while SDH‐activity and muscle fiber type distribution remained unaffected. The ex vivo anabolic response to amino acid enriched buffer was totally blunted in LOWOX‐mice (P=0.039) with an increase of MPB (P=0.036). In LOWOX‐mice, EDL‐force production was decreased at lower stimulation frequencies (P=0.038) and fatigue resistance was diminished (P=0.023).

Conclusions

Reduced dietary intake of vitamins A/E/B6/B12/Folate/selenium/zinc has major impact on muscle health as shown by decreased force production and PA, without effects on muscle mass. Moreover, muscle protein turnover was affected. The reduced FCSA in combination with lack of effect on SDH‐activity suggests that such dietary reductions cause muscle fiber atrophy and reduced muscle oxidative capacity resulting in reduced muscle fatigue resistance.
Increased intracellular calcium is a mediator of heat stress induced adipocyte hypertrophy
Authors: Ajuwon Kolapo M , Qu Huan
Published: November 2023

Exposure of porcine preadipocytes to heat stress (41.5°C) during differentiation results in increased intracellular triglyceride storage, characterized by elevated expression of glyceroneogenic genes such as phosphoenolpyruvate carboxykinase (PCK1) and glycerol kinase (GK), compared to cells differentiated at control (37°C) temperature. However, the molecular mediators of this response are unknown. Therefore, we investigated the role of calcium in this response. Compared to cells differentiated at control temperature, heat stress results in increased intracellular calcium release from the endoplasmic reticulum(ER) in response to ionomycin (2μM), suggesting that heat stress may promote increased ER calcium storage. To determine wether calcium plays a role in the increased lipid storage during heat stress, cells were treated with or without the calcium chelator, FURA2‐AM, during differentiation. Treatment with FURA2‐AM resulted in reduced lipid storage in response to heat stress, suggesting that calcium may play a role in the stress‐induced lipid storage. We also investigated the potential role of calcium channels TRPV1 and TRPV3 because of their implication in heat sensing mechanism. The use of inhibitors to both channels, AMG 9810 (5μM) and DPTHF (100μM), resulted in reduction in intracellular calcium and triglyceride concentrations. Therefore, the TRP channels may play a role in the elevated intracellular calcium concentration in adipocytes during heat stress. The implication of the increased ER calcium release on ER stress and lipid synthesis is still being investigated.
Cat's choose 30% of their calories as protein which is associated with both body composition and circulating metabolites
Authors: Jewell Dennis Edward , Vondran Jodi C. , Vanchina Melissa A.
Published: November 2023

As in other mammals, domestic cats have specific taste preferences that can be influenced by macronutrient composition and physical characteristics of food as well as the presence or absence of specific taste imparting compounds (palatability enhancers). This study investigated the relationship of body composition and food choice and that choice on subsequent metabolomic shifts. Experimental foods were prepared with similar palatability for a population (as evaluated by the ratio of the test food to the total daily food consumed [grams test/(grams test+grams control)] through manipulations of factors independent of macronutrients. These foods with similar palatability preference for the population as a whole were then investigated for intake choice with 27 cats with varying adiposity (mean 24.4% fat, min 7.2%, max 32.2%). Four completely balanced foods were available to the cats at all times. The four foods were: a) a food with 42% of calories from protein, b) 43% calories from fat, c) 52% calories from carbohydrate and, d) intermediate levels of calories from protein, fat and carbohydrate. Cats chose 30.3% of calories from protein (Std Dev 4%, min 24% max 38%). Body composition was associated with the choice of percent calories as protein (p=0.008); lean body mass had a linear negative association (p=0.02), as did fat mass (p=0.04) while there was a positive association for the interaction (p=0.02). Overall cats with high body lean and high body fat showed the highest preference for dietary protein. There was a positive association between the percent of calories as protein and urea cycle metabolites (arginine, n‐acetyl arginine, n‐delta ornithine) while increasing calories from protein were negatively associated with specific metabolites in the TCA cycle (isocitrate, alpha ketoglutarate). There was a negative correlation between percent of calories consumed as protein and that consumed as carbohydrate (r=0.84; p<0.001) with only a slight relationships between either protein or carbohydrate calories and percent of calories consumed as fat (p≥ 0.1). However, there was a positive relationship between protein intake and a number of long chain unsaturated fatty acids (oleate, arachidate and docosapentaenoate). In summary, when given the opportunity to choose between foods which had similar palatability cats chose to consume 30.3% of their calories as protein and had specific preferences related to body composition. These intake preferences were associated with metabolic changes reflecting metabolic shifts.

Support or Funding Information

Hill's Pet Nutrition, Inc.
Transgenerational Effects of High Salt Diet During Gestation
Authors: Peres Maria Angélica , de Oliveira Ivone Braga , Furukawa Luzia Naoko Shinohara , Heimann Joel
Published: November 2023

Introduction

The influence of high salt intake during gestation on the offspring was observed in several studies. It is also known that a phenotype can be transmitted from one generation to the next one without alterations in the DNA sequence. Based on this knowledge, we hypothesized that the effects of high salt ingestion during pregnancy can be transmitted to generations F2 and F3. Therefore, gene expression and DNA methylation of some components of RAS system as well as gene 11beta‐hydroxysteroid dehydrogenase (11βHSD), involved in glucocorticoid conversion, were evaluated.

Material and Methods

Female Wistar rats were fed with normal (N) or hight (H) salt diet during pregnancy. Afterwards, all animals were fed with N, until the third generation. Body and organ mass, gene expression (RT‐qPCR) and methylation (High Resolution Melting – in progress), were evaluated in three generations of males.

Results

data presented as mean±SEM. Table

No differences between H and N were observed in AT1a, ACE (angiotensin conversion enzyme) and HSD11β2 DNA methylation in the kidney of F1.

Discussion

High salt intake during pregnancy has a transgenerational effect on kidneys, testis and white adipose tissue growth. The gene expressions that were evaluated were influenced by the maternal dietary salt content in a variable manner in the offspring along three generations.

Support or Funding Information

This study was supported by FAPESP (2012/22415‐5, 2012/22801‐2)

Variable \ Generation
FI
F2
F3

Body weight (g)
N
458.8±8.64
492.7±11.03
455.1±11.75

H
438.1±8.54
464.3±7.86*
475.4±8.54

Right kidney mass (g/100g BW)
N
0.32±0.004
0.33±0.008
0.33±0.005

H
0.36±0.007*
0.35±0.004*
0.34±0.005*

Left kidney mass (g/100g BW)
N
0.33±0.007
0.32±0.007
0.33±0.005

H
0.36±0.007*
0.34±0.005*
0.34±0.003*

Left testis mass (g/100g BW)
N
0.35±0.01
0.36±0.008
0.39±0.001

H
0.39±0.01*
0.39±0.009*
0.36±0.007*

Retro peritoneal fat mass (g/100g BW)Kidney gene expression
N
0.50±0.10
0.76±0.05
0.68±0.006

H
0.82±0.06*
0.73±0.06*
0.88±0.006*

AT1a
N
0.82±0.27
1.28±0.39
0.93±0.10

H
1.80±0.43*
1.33±0.24
1.06±0.15

AGT
N
0.85±0.21
1.00±0.11
1.60±0.17

H
1.51±0.24
0.87±0.10
0.96±0.23*

HSD11β2
N
1.03±0.43
1.03±0.11
0.97±0.16

H
1.65±0.28*
1.33±0.22
1.11±0.18

difference between groups (N vs. H) ‐p≤0.05. Number of animals: F1 (N=9;H=6), F2 (N=12;H=19) and F3 (N=11;H=16).

AT1a=angiotensin II receptor 1a; AGT=angiotensinogen.
Barriers to Nutritional Counseling by a Registered Dietitian for People Living with HIV (PLWH)
Authors: Fleetwood Christina D , Campa Adriana , Martinez Sabrina , Charlesworth Aimee , Alvarado Glenda , Dizon Kimberly , Li Tan , Huffman Fatma G , Baum Marianna
Published: November 2023

Objective

People living with HIV (PLWH) face multiple barriers to access treatments, experiencing unmet medical and ancillary service needs. The purpose of this study was to determine the barriers that affect adequacy of dietary intake and impede attendance and compliance with nutritional counseling sessions with a Registered Dietitian (RD).

Methods

After consent, a subsample of the Miami Adult Studies on HIV (MASH) cohort consisting of 130 consecutively recruited HIV‐positive study participants completed a survey regarding nutritional services, barriers to services and compliance with visits. In addition, healthcare workers that provide services to PLWH, completed a survey to determine barriers preventing these caregivers from making referrals for dietary counseling. Student t‐tests and chi‐square were used to analyze data on barriers. Spearman rank correlations were used to estimate associations between identified barriers and other categorical outcome variables that affect attendance with an RD.

Results

Of the 130 participants surveyed, 46.2% reported nutritional counseling with an RD in the previous 12‐months. Although 75.4% of all participants have seen an RD in the past, more than half of them (53.8%) were not receiving nutritional services. The most trusted sources (68.5%) of nutritional information were equally the Physician and RD. HIV‐positive participants were more frequently interested in learning about the following topics: weight concerns (73.1%), shopping healthy (72.3%) and on a budget (70.3%). The most identified barriers to participating were difficulty in keeping appointments (33.8%), location (24.6%) and lack of referral to services (23.8%). Barriers that the participants identified towards nutritional counseling were highly correlated with each other. Barriers affecting the referrals to RD's from HIV health professionals included that the patient was not interested, education level, and patient not requesting the services.

Conclusions

Our findings suggest that the majority of PLWH in Miami‐Dade County have received some nutritional counseling with an RD at some point after HIV diagnosis. The most commonly identified barriers to access nutritional care can be overcome by providing education to healthcare providers and patients, facilitating transportation, and providing locations closer to the places where they are receiving medical treatment to make it more convenient.
No Differential Effect of Beverages Sweetened with Fructose, High‐fructose Corn Syrup, or Glucose on Systemic Inflammation in Healthy, Normal Weight or Obese Individuals: a Randomized Controlled Trial
Authors: Kuzma Jessica , Cromer Gail , Hagman Derek , Breymeyer Kara , Roth Christian , Foster‐Schubert Karen , Holte Sarah , Weigle David , Kratz Mario
Published: November 2023

Consumption of sugar‐sweetened beverages (SSBs) is associated with type 2 diabetes and cardiovascular disease, both independently and through increased body weight. Chronic, low‐grade inflammation is associated with these diseases and with obesity. Based on observational data suggesting SSBs promote inflammation, we conducted a randomized, controlled, double‐blind cross‐over intervention to determine whether fructose vs. glucose vs. high‐fructose corn syrup (HFCS)‐sweetened beverages differentially influence biomarkers of systemic inflammation. Based on rodent data showing that fructose triggers intestinal permeability, we also assessed whether SSBs influence biomarkers of gut barrier function. Subjects (24 healthy adults) consumed a standardized diet ad libitum for three 8‐day dietary periods while drinking four servings per day of beverages sweetened with fructose, glucose, or HFCS (55% fructose, 45% glucose), comprising 25% of estimated calorie requirements (consumption mandatory). On average, subjects consumed 116% of their estimated calorie requirement while drinking these beverages, with no difference in total energy intake between groups. C‐reactive protein (CRP) concentrations did not change significantly while subjects consumed the beverages (p=0.353). At the end of the fructose, HFCS, and glucose phases, neither CRP (p=0.403) nor interleukin‐6 (p=0.933) differed from each other significantly. The lactulose‐mannitol ratio suggested that intestinal permeability decreased following the HFCS period compared to the glucose and fructose periods (p < 0.001), while no difference was seen in zonulin (p=0.366) or lipopolysaccharide‐binding protein (p=0.387), a measure of lipopolysaccharide exposure. This well‐controlled intervention demonstrates that medium‐term consumption of fructose‐ vs. HFCS‐ vs. glucose‐sweetened beverages does not differentially affect biomarkers of systemic inflammation or intestinal permeability.

Support or Funding Information

Supported by NIH grant R21HL108257 (to M. Kratz), NIH grant P30 DK017047 (University of Washington Diabetes Research Center), and NIH grant P30 CA015704 (FHCRC Cancer Consortium Cancer Center Support Grant). J.N. Kuzma was supported in part by grant R25CA094880 from the National Cancer Institute
Changes in the Expression of Cell Surface Markers in Spleen Leukocytes in a Murine Model of Frequent Sucralose Intake
Authors: Delgado Marcela Sánchez , Estrada José A. , Contreras Irazu
Published: November 2023

Sweeteners are used in many food products like additives. In the last few years, ingestion of both nutritive and non‐nutritive sweeteners (NNS) has considerably increased. In addition, the general population has also increased its NNS intake, without a specific nutritional recommendation for doing so, as opposed to people suffering from non‐communicable diseases such as obesity, diabetes mellitus and hypertension. Chronic intake of these compounds may indirectly affect cellular metabolism. The main defense for the organism is the immune system, in which energy availability has an impact on cellular populations and their activation capacity. For this reason, it is necessary to determine the effect of frequent intake of NNS on the effector capacity of immune cells. The objective of this research is to determine changes in leukocyte populations and their expression of surface markers in the spleen of mice supplemented with commercial sucralose. 14 week‐old BALB/c mice ingested commercial sucralose ad libitum in daily water for 6 weeks. Leukocytes were isolated from spleen and we performed flow cytometry using the surface markers CD3, CD19, CD16 and CD14 for leukocyte subpopulations, along with CTLA‐4, PD1, CD40L, MHC‐I, MHC‐II, CD80 and CD86. Our results show a decrease in the frequency of B lymphocyte population and T lymphocytes in comparison to the control group. In B and T lymphocytes the analysis of co‐stimulatory molecules show a lower frequency compared to the control group. The immune response depends of the differentiation and activation of cellular populations. We hypothesized that chronic ingestion of commercial sucralose might be affecting the immune response by modifying the frequencies of cellular populations, as well as the expression of co‐stimulatory and inhibitory molecules such as MHC‐I, MHC‐II and PD1, by decreasing the ability of co‐stimulation between B an T lymphocytes, with a probable effect on the immune response. It is necessary to further determine if sucralose intake affects the efficiency of the immune response.
CHRONIC CONSUMPTION OF NON‐CALORIC SWEETENERS IN CARBONYLATED PROTEIN CONCENTRATIONS FROM BLOOD LYMPHOCYTES OF CD1 MICE
Authors: MARTINEZ BEATRIZ ELINA , Escoto Herrera Jorge A , Iniesta Lucia García , RAMOS Roxana Valdés , López Ana Laura Guadarrama
Published: November 2023

The consumption of non‐caloric sweeteners as a strategy to reduce energy intake has recently increased. To our knowledge, there's not studies about protein carbonylation in lymphocytes based in sweeteners consumption. The aim of this study was to compare the effect of chronic consumption of non‐caloric sweeteners in carbonylated protein concentrations from blood lymphocytes of CD1 mice. We used 32 63‐day old CD1 mice, divided into 4 groups: Control (Cl), Sucrose (Suc), Sucralose (SUC), and Stevia (STE). 10g of SAC and 1g of SUC and ST were diluted in 240mL of ultrapure water, mice drank and fed ad libitum. Sweeteners were administrated for 6 weeks, after that, the mice were sacrificed by cervical dislocation and blood obtained by cardiac puncture. Lymphocytes were isolated and 2–4 Dinitrophenylhydrazine assay for quantification of protein carbonylation was performed. There was a significant difference between study groups (F=4.93, p<0.01). Carbonylated protein concentrations in SUC was significantly higher than SAC (Bonferroni test p<0.001) but ST was significantly lower than SUC (Bonferroni test p<0.01) indicating that chronic consumption of SUC may be was associated with increased protein carbonylation in blood lymphocytes.

Support or Funding Information

UAEM
In vitro antimalarial activity of microbial metabolites from mango tannins (Mangifera indica L.)
Authors: Venancio Vinicius Paula , Abrão Lailah Carvalho , Kim Hyemee , Talcott Stephen T , Mertens‐Talcott Susanne U
Published: November 2023

Malaria is a life‐threatening disease caused by Plasmodium parasites that infect humans through the bite of the female Anopheles mosquito. Although malaria is a preventable and curable disease, it is still responsible for many deaths‐mostly children and pregnant women‐especially in Africa. The absence of effective vaccines and the ever‐increasing resistance to common drugs has led to the development of alternative treatment and prevention strategies. The purpose of this study was to evaluate the antimalarial activity of a polyphenolic mango extract (var. Ataulfo, 2.5 – 40 mg/L GAE) and pyrogallol‐derivatives (0.5 – 50 mg/L) in P. falciparum parasites by the Pico Green microfluorometric DNA‐based assay. Two chloroquine‐resistant (Dd2 and FCR‐3) and one chloroquine‐sensitive (HB3) strains were used in this study. Synchronized cultures consisting of early (rings) or late stage (trophozoites) at 1% parasitemia and 2% hematocrit were allowed to grow in the presence of mango and pyrogallol compounds in 96‐well plates for 48 hours in CO2 at 37 °C. Chloroquine (0.2 μM) was used as positive control. Growth inhibition was assessed by the relative fluorescence intensity of treatments compared to the untreated group. Mango reduced the DNA content at 20 mg/L and 50 mg/L for HB3 rings and trophozoites, respectively; pyrogallol presented the same activity at 10 mg/L for both parasite stages. For the Dd2 strain, the parasite growth inhibition was observed in the presence of mango at 10 mg/L for both rings and trophozoites; for pyrogallol, the inhibition was observed at 0.5 mg/L and 1 mg/L for rings and trophozoites. For the FCR‐3 strain, mango inhibited rings at 20 mg/L and trophozoites at 10 mg/L; pyrogallol showed the same effect at 10 mg/L for both stages. These results suggest mango polyphenols and pyrogallol exhibit antimalarial activity.

Support or Funding Information

Texas A&M University and the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES) Research Grant
High Caffeine Consumption and its Association with Lower Intakes of Antioxidant Micronutrients, and with Higher Food Insecurity in People Living with HIV in the Miami Adult Studies on HIV (MASH) Cohort
Authors: Ramamoorthy Venkataraghavan , Campa Adriana , Rubens Muni , Sales Sabrina , Fleetwood Christina , Stewart Tiffanie , Hatsu Irene , George Florence , Khan Hafiz , Liuzzi Juan P. , Baum Marianna
Published: November 2023

Objective

High caffeine consumption has been associated with decreased appetite and weight loss in people living with HIV (PLWH). This could adversely affect levels of antioxidant micronutrients consumed through diet. This study examines the relationship between caffeine consumption and dietary intake of antioxidant micronutrients such as vitamins A and C, zinc and selenium, and with food security scores.

Methods

After consenting 130 PLWH on stable antiretroviral therapy, who were recruited from the Miami Adults Studies on HIV (MASH) cohort, caffeine intake was measured using standardized questionnaires and 24‐hour dietary recalls were used to estimate food intakes. The USDA Food Security Questionnaire was collected from each participant to assess food security scores. Nutribase‐9 was used for estimating the nutrient content of the foods consumed. SPSS version 21 was used for statistical analyses. More than 250 mg of caffeine intake or the equivalent of 3–4 cups of coffee daily was considered the cut‐off point for high consumption. This study was approved by the Florida International University Institutional Review Board.

Results

Mean age of this group was 47.9 ± 6.4 years, 60.8% were men, and 75.4% were Black. Mean caffeine intake was 337.63 ± 304.97 mg/day, and 22% reported inadequate caloric intake for their requirements; 76.9% reported deficient intakes of vitamin A, 64.6% of vitamin C, 84.6% for zinc and 50.8% for selenium. Compared to participants who used caffeine with moderation, intakes of vitamin C (110.38 ± 141.475 vs 61.68 ± 71.91 mg/day, P=0.029), zinc (9.13 ± 1.774 vs 4.64 ± 1.753 mg/day, P=0.027) and selenium (76.61 ± 5.797 vs 45.27 ± 5.837 μgm/day, P=0.029) were significantly lower in participants who consumed high levels of caffeine per day. High food insecurity was associated with high caffeine consumption (β=1.106, P=0.042) and remained significant after adjusting for covariates.

Conclusion

High caffeine intake adversely affected the consumption of antioxidant nutrients in PLWH in the MASH cohort and was associated with food insecurity. Further studies are needed to estimate the long term adverse effects of these dietary deficiencies on metabolic outcomes in HIV disease progression and quality of life in this population.

Support or Funding Information

Funded by NIDA and FIU Graduate School Dissertation Year Fellowship.
Effect of Age on Vitamin D in Serum of Male Rhesus Monkeys
Authors: Preston Alan M , Rodriguez‐Orengo Jose , Ayala‐Torres Sylvette
Published: November 2023

Limited studies have been carried out on blood levels of vitamin D in non‐human primates. Herein we present data using the preferred vitamin D indicator, serum 25‐OH vitamin D, collected from 56 male Rhesus monkeys from 1 yr to 31 yrs of age. To minimize number of variables, all sampling was done from May thru September, all animals were housed in specific pathogen‐free cages and fed Tekland NIB Primate diet (containing 8000 IU/kg Vitamin D3. Serum 25‐OH vitamin D was quantified using HPLC. Quantitative circulating levels were below those we previously observed in female Rhesus monkeys however a clear pattern in blood levels of males developed starting with values from the 1–5 yr old group which increased progressively thru the 6–10 yr old group with a slight decline in the 11–15 yr old group, a greater decline in the 16–20 yr old group to the lowest values in aged monkeys (21–31 yrs). Most studies in humans indicate that vitamin D levels do not change with age however our study did not measure dietary intake which could possibly explain this discrepancy.

Support or Funding Information

Work reported here was partially supported by the Caribbean Primate Research Center Grant number 5 P40 OD12217.
Vitamin A status affects the Body Mass Gain and Hepatic Gene Expression in Rats fed a High‐Fat Diet
Authors: Kuang Heqian , Li Yang , Eastep Jennifer , Chen Guoxun
Published: November 2023

Introduction

Vitamin A (VA) plays a role in variety of physiological functions from development to metabolic homeostasis. High‐fat (HF) diet has been shown to cause the development of metabolic diseases such as obesity and type II diabetes. We hypothesize that VA status may affect the gain of body mass in rats fed a HF diet and regulate the expression levels of genes involved in hepatic glucose and fatty acid metabolism.

Methods

Sprague Dawley (SD) rats at weaning were fed either a HF‐VA sufficient (HF‐VAS) or HF‐VA deficient (HF‐VAD) diet for 8 weeks. The HF‐VAS and HF‐VAD diets are isocaloric. Their body mass and food intake were recorded once a week. The expression levels of hepatic genes for glucose and fatty acid metabolism, and insulin signaling pathway were determined via immunoblotting.

Results

From week 6, body mass of HF‐VAD rats was significantly lower than that of VAS group. The total food intake was similar between VAD and VAS groups. The liver mass and white adipose mass of HF‐VAD group were also significant smaller than that of HF‐VAS group. The hepatic expression level of Cytochrome P450 26A1 protein in HF‐VAS group is higher than that in HF‐VAD group, suggesting reduced retinoic acid production. For glucose metabolism, the HF‐VAD rats had higher glycogen synthase 3α (GSK‐3α) and lower GSK‐3β expression levels than the HF‐VAS rats did. The expression levels of glucokinase and cytosolic form of phosphoenoylpyruvate carboxykinase in HF‐VAS group are higher than that in HF‐VAD group. Proteins levels of acetyl‐CoA carboxylase (ACC), phospho‐ACC, fatty acid synthase, and ATP citrate lyase in fatty acid synthesis, are similar between two groups. The levels of insulin receptor beta subunit and insulin receptor substrate 1 are increased in HF‐HF‐VAD rats, while that of insulin receptor substrate 2 are decreased. The expression level of mitogen‐activated protein kinase of HF‐VAD group was higher than that of HF‐VAS group.

Conclusions

VA status reduces the body mass gain in rats fed a HF diet. In addition, the expression levels of genes for glucose metabolism and insulin signaling pathway are also altered by the development of VA deficiency in these rats. These results support our previous observations that VA status and its metabolism affect the hepatic expression levels of insulin‐regulated genes in rats fed the diets with normal fat content. More studies are needed to determine the underlying mechanisms for these changes.

Support or Funding Information

Internal funding
Zinc Supplementation, Beta Cell Function, Insulin Secretion, and Insulin Resistance in Black and White Female Adolescents
Authors: Lobene Andrea J , Kindler Joseph M , Pollock Norman K , Laing Emma M , Lewis Richard D
Published: November 2023

Pubertal growth is a period of time characterized by increasing insulin resistance. Zinc is an essential micronutrient required for the maturation and storage of insulin in pancreatic beta cells, and its effect on insulin concentrations and insulin resistance in growing children is uncertain. The objective of this study was to determine the influence of zinc supplementation on beta cell function, insulin secretion (C‐peptide), and insulin resistance in healthy female children. A secondary objective was to determine if the effect of zinc supplementation on these outcomes differs by race. Healthy non‐Hispanic black and white females ages 9–11 y (n=147) were randomly assigned to receive either a daily oral zinc supplement (ZN; 9 mg elemental zinc; n=75) or an identical placebo (PL; n=72) for 4 weeks. Fasting insulin, glucose, and C‐peptide were assessed at baseline and 4 weeks, and insulin and glucose values were used to calculate homeostatic model assessment (HOMA2) of beta cell function (%β), insulin resistance (IR), and insulin sensitivity (%S). A mixed ANOVA design, with time as the within‐subjects factor and treatment and race as between‐subjects factors, was used to assess changes in these outcome measures. All values were non‐normally distributed, and were log transformed prior to analyses. At baseline, the overall mean (± SD) BMI‐for‐age percentile was 75.1 ± 26.1, fasting glucose, 86.1 ± 7.6 mg/dl, fasting insulin, 192.2 ± 110.8 pmol/L, and C‐peptide, 2.1 ± 1.1 ng/ml. There were no baseline differences between the ZN and PL groups for any of the variables. An interaction that approached significance was found for C‐peptide, such that an increase in C‐peptide was observed in the PL group, but not the ZN group, in blacks, but not in whites (p=0.075). No changes were observed in HOMA2‐%β, HOMA2‐IR and HOMA2‐%S for any race‐treatment group after 4 weeks. Our results suggest that zinc supplementation may attenuate an increase in insulin secretion experienced by black, but not white, premenarcheal females. Additional trials, including dose‐response trials, could provide greater insight into the effects of zinc on insulin secretion and the differential responses by race.

Support or Funding Information

National Institutes of Health (NIH grant R03 HD54630)
Dual Action of Zinc in Bone and Vascular Calcification
Authors: Kwun In‐Sook , Shin Mee‐Young
Published: November 2023

Osteogenic calcification is a normal physiological process in bone tissue, anywhere else like the blood vessel, it is a pathological process which is so‐called calcification paradox. The retarded skeletal growth in animals and man is the major sign of zinc inadequacy, which implies the involvement of zinc in osteogenic calcification.

Our previous studies showed that inadequate zinc level in osteoblasts decreased osteogenic activities by decreasing extracellular matrix calcification, mainly through the inhibition of alkaline phosphatase (ALP) expression and activity, and by down‐regulation of osteoblast differentiation marker genes (osteopontin, osteocalcin and collagen type I), which is regulated through the reduced and delayed expression of bone‐specific transcription factor Runx2. Therefore, it seems zinc can be a stimulator in bone formation which includes osteogenic bone matrix calcification.

Meanwhile, there is emerging evidence that zinc can protect against vascular damage and therefore may decrease the incidence of atherosclerotic plaque calcification. Our recent study using rat aortic smooth muscle cell line (A7r5) and rat primary vascular smooth muscle cells (pVSMC) showed that inadequate zinc level in VSMCs increased VSMC calcification and this calcification is paralleled with VSMC apoptotic pattern, rather than osteogenic calcification mode. In mice study using atherosclerotic model (ApoE null mouse), zinc deficiency promotes aorta Ca and P deposition (vascular calcification), while it decreases Ca and P contents in femur (bone calcification). The findings from our in vitro (osteoblasts and VSMCs) and in vivo (atherosclerotic ApoE null mouse) studies suggest that zinc may have a reciprocal role for calcification in bone (hard tissue) and blood vessel (soft tissue). It can be suggested that zinc may have a potential for therapeutic strategy for atherosclerosis as well as osteopoprosis, both of which involve the process of calcification.

Support or Funding Information

(Supported by the National Research Foundation of Korea, NRF‐2011‐0014535)
3D Printing in Anatomical and Palaeontological Teaching and Research
Authors: Adams Justin Winfield
Published: November 2023

Widespread commercialisation of additive manufacturing (3D printing) machinery and non‐invasive 3D data capture devices is increasingly allowing academics access to these technological advancements. As translating 3D datasets into 3D prints becomes feasible for scientists and educators from non‐engineering disciplines, it is critical to consider the potential benefits and challenges for incorporating the technology into research and teaching. This presentation covers several of these key benefits and challenges of current 3D data capture and printing technologies, and summarises the recent published research on their use in the anatomical and palaeontological sciences. The most significant challenges to using 3D printing involve financial considerations (e.g., equipment choice, funding, facilities, technical support), practical decisions (e.g., choice of materials, fidelity of prints as research materials), and engagement with the larger scientific community (e.g., intellectual property, data sharing). The benefits of 3D printing include the baseline ability to preserve, duplicate, and make accessible rare, fragile, or important anatomical and palaeontological specimens; to the production of customised research and teaching materials to expand data collection, student learning and public outreach. Within anatomy and allied discipline education, initial studies of student use of 3D prints point to their potential benefit, but will ultimately require careful evaluation (like all novel teaching tools) to assess their effectiveness and impact on student learning outcomes.
Virtual Surgical Planning and Applications in Plastic and Craniomaxillofacial Surgery
Authors: Basel Sharaf , Matsumoto Jane , Lachman Nirusha , Bite Uldis
Published: November 2023

Advanced plastic and craniomaxillofacial reconstruction is a multidisciplinary field, requiring communication and accurate planning among various specialists including oncologic surgeons, plastic surgeons, and radiologists. Complex defects may present in pediatric congenital deformities, after trauma or oncologic surgery. Restoration of form and function require precise planning among the various specialists involved and accurate execution of the surgical plan. Often, considerations for bony and soft tissue requirements make the reconstruction more challenging. Virtual surgical planning using computed tomographic imaging, 3D surface imaging technology and computer‐aided design allow the surgeons to perform virtual surgery and generate templates and cutting guides that allow for the precise and expedient recreation of the plan in the operating room. The author presents cases highlighting various reconstructive requirements where virtual planning was used for advanced plastic and craniomaxillofacial reconstruction to achieve precise reconstruction.

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None
Inter‐competetion and conversion among TNF‐α‐induced signaling complexes determine the outcomes of TNF‐α signaling
Authors: Yang Zhang‐Hua , Wu Xiao‐Nan , Wang Xuekun , He Peng , Hong Mao , Han Jiahuai
Published: November 2023

Inflammatory gene induction, apoptosis and necroptosis are major cellular responses of the cells exposed to tumor necrosis factor (TNF)‐α. Here we systematically investigated inter‐relationships among the key signaling molecules that control these cellular processes to uncover the molecular mechanism underlying the inter‐competetion and conversion among TNF‐α induced signaling complexes that elicit multiple alternative outcomes of TNF‐α in a murine system. By taking advantage of quantitative mass spectrometry, we found that the amount of Receptor interacting protein 1 (Rip1) and Tradd recruited to the TNF receptor 1 (TNFR1) complex is the initial determinant factor that governs the cell's decision to undergo cell death or not. Rip1 in complex I downstream of TNFR1 engagement not only initiates the signaling for inflammatory gene expression but also inhibits Tradd‐dependent recruitment of Fadd and caspase‐8 that can lead to rapid apoptosis. When there is no Rip3 or Mlkl in cells, elimination of the ubiquitination of Rip1 leads to formation of complex II, which triggers slower apoptosis than occurs in the absence of Rip1. Complex II can be converted to the necrosome when there are sufficient Rip3 and Mlkl. Both caspase‐8 and Flip can function in the necrosome to suppress necroptosis. Recruitment of Rsk1 to the necrosome is another layer of security to process necroptosis because Rsk1 inactivates caspase‐8 by phosphorylation. Thus, Rip1 is the central controller for cells to have induction of inflammatory genes and/or cell death; the interplay among Rip3, caspase‐8, Flip and Rsk1 determines whether the cells undergo apoptosis or necroptosis.

Support or Funding Information

This work was supported by the National Basic Research Program of China (973 Program; 2015CB553800), the National Scientific and Technological Major Project (2013ZX10002‐002), the National Natural Science Foundation of China (91429301, 31420103910, 31330047 and 31221065), the Hi‐Tech Research and Development Program of China (863 program; 2012AA02A201), the 111 Project (B12001), the Science and Technology Foundation of Xiamen (No. 3502Z20130027), the National Science Foundation of China for Fostering Talents in Basic Research (Grant No.J1310027) and the Open Research Fund of State Key Laboratory of Cellular Stress Biology, Xiamen University.
Cathepsin Localization and Activity on the Intestinal Epithelial Barrier of Rats following Hemorrhagic Shock and Reperfusion
Authors: Courelli Asimina S. , DeLano Frank , Schmid‐Schonbein Geert W.
Published: November 2023

Damage to the intestinal epithelial barrier during hemorrhagic shock results in tissue destruction and increased permeability. Although the downstream effects of damage to the intestinal epithelial barrier have been investigated, the mechanism behind the increase in permeability has not been elucidated. Previous studies have suggested a proteolytic source for the degradation, but the spectrum of proteolytic enzymes in the intestinal tissue is very broad. We employed mass‐spectroscopy based proteomic analysis of a jejunal intestinal sample obtained from a young adult male Wistar rat (flushed with PBS to remove luminal contents) and found that the enzymes with the greatest relative abundance among the proteolytic enzymes were cathepsin–B, –C, and –Z. We examined cathepsin localization profiles using rat tissue from normal control (NC) Wistar rats and rats that underwent ischemia (at a blood pressure of 35 mmHg) followed by 2 hours of reperfusion (IR). Immuno‐histochemistry (IHC) was used to determine the cathepsin localization profiles. In the case of NC, comparison of cathepsin–B, –C, and –Z localization profiles of the intestinal epithelial barrier between rats (in our study) and humans (Swedish Human Protein Atlas) revealed a significant degree of similarity. Western Blots and gel zymography were conducted to determine changes in cathepsin concentration and activity respectively. Western blots showed a decrease in cathepsin levels, while gel zymography showed no changes in cathepsin activity between NC and IR groups. However, cathepsin activity per mole of total cathepsin (expressed by the ratio between the gel zymography and western blot bands) was higher in the case of IR, supporting the potential role cathepsins play in causing damage to intestinal tissue during hemorrhagic shock, leading to increased intestinal permeability.

Support or Funding Information

Supported by HL 10881.
Cellular stress response 1 regulates RNA splicing of epidermal growth factor receptor and platelet‐derived growth factor receptor through inactivation of splicing factor 3A3
Authors: Luo Jianhua , Zuo Ze‐Hua , Yu Yan P
Published: November 2023

Cellular stress response 1 (CSR1) is a tumor suppressor gene that plays an important role in regulating cell death. In this report, we showed that the N‐terminus of CSR1 interacts with splicing factor 3A, subunit 3 (SF3A3). The SF3A3 binding motif was identified in the region of amino acids 62–91 of CSR1 through cell‐free binding analyses. The interaction between CSR1 and SF3A3 led to migration of SF3A3 from nucleus to cytoplasm. The cytoplasmic redistribution of SF3A3 significantly reduced the splicing efficiency of epidermal growth factor receptor and platelet‐derived growth factor receptor. Induction of CSR1 or down‐regulation of SF3A3 also significantly reduced the splicing activity of oxytocin reporter gene both in vivo and in vitro. Mutant CSR1 that lacks the SF3A3 binding motif contained no RNA splicing regulatory activity, while the peptide corresponding to the SF3A3 binding motif in CSR1 interfered with the wild type CSR1 mediated inhibition of RNA splicing. Interaction of CSR1 and SF3A3 is essential for CSR1 mediated cell death. To our knowledge, this is the first report demonstrating that RNA splicing is negatively regulated by redistribution of a splicing factor.

Support or Funding Information

National Cancer Institute (RO1 CA098249)
Induction of Anchorage Independent Growth and Angiogenesis by Tungsten Carbide‐Cobalt Nanoparticles
Authors: Ding Min , Barber Tabatha lynn , Bowman Linda
Published: November 2023

Powder mixtures of nano‐sized tungsten carbide‐cobalt(WC‐Co) have wide applications because of their hardness and toughness. WC‐Cois known to be associated with pulmonary injury and malignant diseases. In this study, we investigated anchorage independent growth and the angiogenic response after exposure to WC‐Co nanoparticles. The results showed that chronic exposure of BEAS‐2B cells with low dose (0.025μg/cm2) WC‐Co significantly induced cell transformation in soft agar. WC‐Co nanoparticles also induced angiogenesis as determined by the chicken chorioallantoic membrane (CAM) assay. Mechanistic studies indicated that WC‐Co nanoparticles stimulatedalteration of miR‐21, PDCD4, AP‐1, NF‐kB, VEGF, AKT and ERKs activities. Inhibition of ERKs with U0126 reversed WC‐Co‐induced PDCD4 inhibition, but inhibition of p38 with SB203580 did not. These results illustrate that exposure to WC‐Co nanoparticles induces acarcinogenic response by activating AKT‐VEGF, and AP‐1‐MAPKs signal pathways and the downstream molecules. This information may be useful for preventing potential damage from nanoparticle exposure in the future.
Moringa oleifera Whole Methanolic Leaf Extract Attenuates Levels of Angiogenic Factors in the Cervix of Preterm Labor Mice Models
Authors: Fitch Morgan , Ray Savannah , Zwetsloot Kevin A. , Mowa Chishimba
Published: November 2023

On average, 1 in 10 births in the United States are born premature, and the leading cause is microbial infection and inflammation. Current treatments have serious side effects and are undermined by the growing multi resistance to anti‐microbial agents. We recently demonstrated a positive feed forward relationship between expression of vascular endothelial growth factor (VEGF), a key angiogenic factor, in mice cervix and inflammation. We also revealed that Moringa oleifera, an ancient medicinal plant, exhibit potent anti‐bacterial (E. coli) and ‐inflammatory activities in vitro and in cervix of the same mice models, respectively. Here, we test whether Moringa oleifera can also modulate expression of VEGF and other angiogenic factors in the cervix of preterm labor mice models. Cervical tissues of pregnant mice were harvested and analyzed using real‐time PCR (qPCR) and Western blot after specific treatments [2 hour pretreatment with Moringa oleifera (4.8 μg/50 μL methanol, oral) followed by 2 hour treatment with lipopolysaccharide (LPS) (100 μg/50 μL 1X PBS, i.p.)]. Other angiogenic factors examined include VEGF receptor 1 (Flt‐1), VEGF receptor 2 (KDR), and endothelin‐1 (End‐1). The results show that Moringa oleifera whole leaf methanolic extract overall down regulates expression of VEGF, KDR, and End‐1 mRNAs, while it had no significant effect on the expression of Flt‐1. Based on these data, we conclude that Moringa oleifera may be used to intercept the positive feed forward loop between angiogenic and inflammatory factors, and thus potentially tone down inflammation and subsequently preterm labor.

Support or Funding Information

Appalachian State University, Office of Student Research.
Reacquisition of E‐cadherin Expression in Metastatic Signet Ring Cell Carcinoma: A Potential Anchor for Metastatic Deposition
Authors: Ma Yihong , Siegal Gene P , Wei Shi
Published: November 2023

E‐cadherin is a cell‐cell adhesion molecule with pivotal roles in epithelial cell behavior. Loss of E‐cadherin leads to disappearance of epithelial characteristics and down‐regulation of migratory and invasive properties of cells, a process known as epithelial–mesenchymal transition (EMT), fundamental to the initiation of metastasis during cancer progression. Signet ring cell carcinoma (SRCC) is an aggressive histologic subtype of adenocarcinoma with a diffuse infiltrative nature. We have previously demonstrated that reduced expression of E‐cadherin is a frequent event in SRCCs of various organ systems. The aim of this study was to examine the expression of E‐cadherin in paired primary and metastatic SRCCs to further explore the potential role of this molecule in metastatic dissemination of this unique tumor type.

Methods

Thirty‐four consecutive cases of SRCC from various organ systems with both primary and metastatic tumoral tissue available were retrieved from the authors’ institution between 2004 and 2014. The intensity of membranous E‐cadherin expression was scored as 3+ (strong; equivalent to adjacent normal epithelium), 2+ (moderate), or 1+ (weak), while no staining was assigned a score of 0. A score of 2+ or less was regarded as reduced expression. Chi‐square or paired t‐test was used for statistical analysis, when appropriate.

Results

Loss or reduction of E‐cadherin expression was a distinguishing feature of primary SRCCs and was observed in 27 (79%) of primary tumor specimens. Interestingly, lost or reduced E‐cadherin expression was less frequently seen in metastatic SRCCs when compared to their primary counterparts, and was only found in 18 tumors (53%) at distant relapse (Odds ratio 3.4; p=0.04). The mean score of E‐cadherin expression of primary SRCCs was significantly lower than that of their metastatic counterparts (2.3 vs. 1.8; p=0.001).

Conclusions

While the reduced E‐cadherin in primary SRCCs supports its role during EMT in tumor progression/metastatic dissemination, the re‐expression of E‐cadherin in SRCC cells at distant relapse sites implies a reversal to their epithelial phenotype (mesenchymal‐epithelial transition; MET) which, in turn, helps tumor cells to anchor and form cohesive metastatic deposits. Thus, opportunistic tumor cells may modulate this process to their advantage to go through EMT‐MET for successful metastasis, a phenomenon potentially regulated by epigenetic mechanisms rather than the accumulation of mutations and other genetic changes which are, by default, irreversible. Further elucidation of this process will help in the development of novel targets for antimetastatic therapies.

Support or Funding Information

None