bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of tomato extract on cytokine profile of human Peripheral Blood Mononuclear Cells (PBMCs)
Authors: Godoy Maria Fernanda , Prieto Ligia Esperanza Diaz , Rebato Esther Nova , Slobodianik Nora Haydee , Insani Ester Marina , Marcos Ascensión
Published: November 2023

Objective

This study was aimed to evaluate the effect of the addition of tomato extract (TE) on cytokine production.

Material and Methods

Human PBMCs (n=7, independent samples) were isolated by Ficoll‐Hypaque gradient centrifugation and cultured with PHA (7μg/ml) and LPS (1.5μg/ml) for 48hs to stimulate cytokine production. TE was achieved by means of saponification and solvent extraction (hexane) in order to obtain the non‐polar compounds naturally included in tomato. A concentration step and subsequent re‐dissolution in ethanol was made (final concentrations <1% in culture cells). In this TE, lycopene and vitamin E are main bioactive compounds. We focused on α‐tocopherol. Four groups were considered: PBMCs + PHA/LPS + 1, 10 or 20μM of α‐tocopherol from TE (PBMC‐1, PBMC‐10 and PBMC‐20 groups, respectively) and PBMCs + PHA/LPS used as the control group (C). IFN‐γ, TNF‐α, IL‐2, IL‐17A, IL‐1β, IL‐4, IL‐13 and IL‐10 were measured in culture supernatants by flow‐cytometry (CBA, EMD Millipore). Statistics were performed using Univariate (LSD Fisher test) and Multivariate Analysis (HCA).

Results

A dose‐dependent response to TE was observed (figure 1). Regarding cytokine production, IFN‐γ and IL‐10 levels were significantly lower (p=0.04 and p=0.02, respectively) in PBMC‐20 than in C. No significant differences were found in the other cytokines. Although TNF‐α showed no significant difference among groups, TNF‐α/IL‐10 ratio increased up to 60% due to the decrease in IL‐10 production in the presence of increased amounts of TE. As observed in figure 2, for a given concentration of TNF‐α, IL‐10 values are lower in the presence of higher amounts of TE (PBMC‐10 and PBMC‐20). Another pro‐inflammatory cytokine, such as IL‐1β showed a similar pattern (figure 3). These findings may indicate a modulation of TE on the pro‐inflammatory cytokine response.

The homeostatic relationship between IFN‐γ and TNF‐α is well‐known. There was a change when TE was added. While in C IFN‐γ/TNF‐α levels were 1.46, in PBMC‐20 this ratio decreased to 0.71 due to a strong decrease in IFN‐γ levels (p=0.01). In this regard, pro‐inflammatory/anti‐inflammatory cytokine ratio was significantly higher (p=0.02) for PBMC‐20 vs. C, showing PBMC‐10 intermediates values, which indicates an anti‐inflammatory state.

Conclusion

In the in vitro system performed in this study, a physiological amount of α‐tocopherol (20μM) and the corresponding level of lycopene from tomato have shown to be able to produce an anti‐inflammatory effect.
The effect of supplemental grape seed extract on pig growth performance and body composition during heat stress
Authors: Smithson Andrew T , Winslow Christopher J , Haufe Thomas C , Ryan Caroline , Gilley Andrew , Strat Karen , Ajlan Reem K.M. , Biggs Morgan , Zhang Zhenhe , Zhao Lidan , Harl Audra , Poole Rebecca , Rhoads Michelle L , Rhoads Robert P , Neilson Andrew Peter
Published: November 2023

Prolonged exposure to high ambient temperature without cooling causes heat stress resulting in altered growth, body composition and metabolic dysfunction. Grape seed extract (GSE) has been shown to reduce inflammation, and improve glucose transport and metabolism. Thus, GSE may be an effective supplement to combat the consequences of heat stress; however this possibility has not been evaluated in a large animal model. The objective of the current study was to examine the effect of GSE supplementation on pig performance and body composition during heat stress. Twenty‐four female pigs (62.3± 8 kg BW) were randomly assigned to a 2X2 factorial experiment; thermal neutral (TN; 21–22°C) or heat stress conditions (HS; 33–34°C) for 7 days and fed either a control or a GSE supplemented diet (12mg/kg body weight). Body temperature (TB), respiration rate (RR) and feed intake (FI) were measured daily. Body composition was measured by dual‐energy X‐ray absorptiometry (DXA). Respiration rate and TB increased in the HS control group compared to the TN control group (p<0.05), however GSE did not alter these parameters compared to control for the duration of the 7 day period. Heat stress decreased FI (P < 0.05). Fasting blood glucose concentrations were approximately 1.5‐fold greater in the control diet compared to their GSE supplemented counterpart (p>0.05) on day 6 of the HS period, but did not differ between groups on day 7 of HS. Body composition analysis indicated bone mineral density, bone mineral content, percent change of fat, and percent change of weight over the study remained unchanged between treatment groups. Lean tissue accretion was 45% greater in TN compared to HS groups (p<0.05). Grape seed extract supplementation does not appear to alter pig growth performance or body composition during heat stress.

Support or Funding Information

John Lee Pratt Endowment Animal Nutrition Program at Virginia Tech
Nicotinamide riboside affects toll‐like receptor 4‐NF‐κB pathway via sirtuin‐dependent and –independent ways in AML12 hepatocytes
Authors: Lee Hee Jae , Yang Soo Jin
Published: November 2023

The objective of this study was to explore the potential role of nicotinamide riboside (NR) on Sirt1/3. The NAD+ precursor NR is a third vitamin B3. Recent studies have suggested that NR shown protective effects for hearing loss, high‐fat induced obesity, and Alzheimer's disease. Mouse hepatocytes were treated with NR (10 μM and 10 mM) for 24 hours. RNA interference (RNAi)‐mediated gene silencing was performed for Sirt1 and Sirt3 knockdown, and then NR was administered. Inflammatory markers and Sirt pathway were analyzed. AML12 cell viability which was reduced by PA was increased by low concentrations of NR treatment. Sirt1 was significantly up‐regulated by NR. The NR treatment led to the up‐regulation of insulin‐sensitizing molecule adiponectin. The levels of CPT‐1 in NR‐treated cells were three times higher than those of CON. Hepatic pro‐inflammatory markers including TNF‐alpha and IL‐6 were decreased in NR‐treated cells. In addition, caspase 1, a component of NOD‐like receptor family, pyrin domain containing 3 inflammasome, was decreased by NR. Sirt1 knockdown triggered hepatic metaflammation. Sirt1 was necessary to show protective effects for inflammation in AML12 NR‐treated cells. Sirt3 knockdown resulted in decreased levels of Sirt1 and Sirt6. NR treatment altered toll‐like receptor 4‐NF‐κB, which was independent of Sirt3. NR decreased level of fatty acid synthase, and these effects were observed in Sirt3 knockdown group. These data demonstrated that NR enhances cell viability, and affects TLR4‐NF‐κB pathway via sirtuin‐dependent and ‐independent ways in hepatocytes.
Iron Storage in Women is Positively Correlated with Aging and BMI Values
Authors: Ma Aiguo , Jia Ru , Ma Xiuting , Sun Yongye , Zhang Huaqi , Ma Yan
Published: November 2023

Iron deficiency occurs commonly in vulnerable groups of women at reproductive age. But many studies have shown that the situation of iron overload in the elderly becomes an increasingly serious issue. In the present study, we aimed to investigate the association of serum ferritin levels with age and body mass index (BMI) and the iron status in middle‐aged women. This cross‐sectional study was carried out on 4625 healthy women in Qingdao who were at the age of 40–70 and had informed consent to participate in the study. Serum ferritin concentrations of all participants were measured by an automated radioimmunoassay analyzer. For female adults, normal serum ferritin is defined by the World Health Organization as 15~150 μg/L, and anemia as <15 μg/L, iron overload as ≥150 μg/L. Among all the participants, 76.25% had a normal iron storage, 19.10% had iron deficiency, and 4.65% had iron overload. The median of serum ferritin concentration in 65–70 years old was nearly two‐fold higher than that in 45–50 yeas old. And in obese people, the median of serum ferritin concentration was significantly higher than that in lean people. Furthermore, serum ferritin was positively correlated with age and BMI, with the correlation coefficient values at 0.376 and 0.156 (both p<0.05), respectively. In conclusion, our results suggest an increasing prevalence of iron overload with age and BMI.

Support or Funding Information

Supported by the National Natural Science fund (No. 81373000)
Baby Friendly Hospital Births and Lactation Consultants Are Associated with State‐Level Breastfeeding Rates
Authors: Taren Douglas , Sinari Shripad
Published: November 2023

Background

Breastfeeding has short and long term protective health implications for both the mother and the child. The breastfeeding rates in the US have not yet met the Healthy People 2020 goals. An annual breastfeeding report card by the CDC identifies five state‐level breastfeeding indicator. It also gives state‐level summary measures for practices that support breast feeding. We used the 2014 CDC breastfeeding report card to help us understand which state‐level support indicators were associated with measures of breastfeeding.

Methods

The breastfeeding outcomes were: percent ever breastfed, percent breastfed for 6 and 12 months, and percent that exclusively breastfed for 3 and 6 months. The support variables were: an overall score of maternity care practices and policies at a birth facility, La Leche leaders per 1000 live births, International Board Certified Lactation Consultants (IBCLC) per 1000 live births, the proportion of state births in baby friendly facilities and percent of infants given formula before 2 days of age. State‐level covariates in our analysis were total population size, percent urban, percent white, median income, percent unemployed, percent below poverty, and percent with high school or greater degree. We conducted canonical correlations to determine which support indicators explained the overall breastfeeding outcomes. We also conducted multivariate multiple regression to adjust for state demographic factors.

Results

The canonical correlation revealed two significant variates for the underlying data. The first variate paired the variable for the percent of women who exclusively breastfed for 3 and 6 months. The percent of infants born at baby friendly hospitals and the percent who were given formula before 2‐days of age were associated with this variate. The second variate paired the percent of infants who were ever breastfed and the percent who were exclusively breastfed for 3 months. The number of IBCLCs per 1000 birth and the percent of infants given formula before 2 days of age were associated with this variate. The multivariate multiple regression model using state‐level covariates indicated that the percent of births in baby friendly hospitals was significantly associated the percent of infants who ever breastfed, the percent who breastfed for 6 months and the percent who exclusively breastfed for 3 and 6 months. Similarly, the number of IBCLCs was significantly associated with the percent who exclusively breastfed for 6 months and who breastfed for 12 months.

Conclusion

We determined that the proportion of births that occurred in baby friendly hospitals and the availability of IBCLCs were associated with promoting positive breastfeeding practices. Community measures such as providing formula before 2‐days of age was related to poorer breastfeeding measures. These results can be used to develop policies to sustain and increase state‐level breastfeeding rates.

Support or Funding Information

None
UGT1A6 polymorphism and plasma bilirubin are associated with antioxidant status in male smokers
Authors: Seo Bo‐Young , Park Eunju
Published: November 2023

This study was conducted to investigate the relationship between plasma total bilirubin level and antioxidant status and to assess the role of UDP‐glucuronosyltransferase (UGT) 1A6 genetic polymorphism, which affects glucuronidation of bilirubin and thus may induce an altered level of bilirubin, in the antioxidant status in male smokers. Plasma antioxidant parameters[TRAP, lipid‐soluble vitamins(Retinol, γ‐, α‐tocopherol, α‐, β‐carotene, and coenzyme Q10), lipid peroxidation, erythrocyte antioxidant enzyme activities(SOD, CAT, and GPx), leukocytes DNA damage], and genetic polymorphisms for UGT1A6 were assessed in 162 male smokers with no evidence of disease and no major risk factors for liver disease. When the subjects were classified into the four groups by quartiles of bilirubin levels, significant differences in TRAP, retinol, γ‐, α‐tocopherol, α‐carotene and coenzyme Q10, activities of erythrocytes antioxidant enzymes(SOD and CAT), and leukocytes DNA damage were found among the groups. In the simple linear regression, the higher level of plasma total bilirubin was associated with the significantly increased TRAP(r2=0.044, p<0.001) and α‐tocopherol(r2=0.092, p<0.001). Additionally, the levels of plasma total bilirubin, TRAP, retinol, γ‐, α‐tocopherol, and coenzyme Q10, activities of SOD and CAT, and leukocyte DNA damage depend on variations of UGT1A6 genotypes(homo+hetero), when AST and ALT were adjusted. These results suggested that the increased plasma total bilirubin within normal range may be accompanied with enhanced physiological antioxidant status in healthy male smokers. Also, the presence of variant UGT1A6 genotypes seem to be associated with altered antioxidant status assumedly mediated by plasma total bilirubin concentration.
Functional Vitamin B12 Status in Lactating Women Living in Rural Indonesia Assessed by Using Dried Blood Spot Methylmalonic Acid Analysis
Authors: Schroder Theresa H , Diana Aly , Quay Teo AW , Gibson Rosalind S , Houghton Lisa A , Lamers Yvonne
Published: November 2023

Background

Low vitamin B12 (B‐12) status in breastfeeding women has been associated with infant B‐12 inadequacy and adverse health outcomes in mothers. Populations in resource‐poor settings are at increased risk for B‐12 deficiency. Dried blood spots (DBSs) are a convenient and minimally invasive tool to collect biological samples in field studies. We have recently developed a method to quantify methylmalonic acid (MMA), the most specific functional biomarker of B‐12 status, in DBSs. The aim of this study was to determine the risk of low B‐12 status among lactating women living in rural Indonesia by assessing DBS MMA in a convenience sample and comparing the results to a reference group of healthy, non‐lactating women.

Methods

Apparently healthy, lactating women living in Sumedang district of West Java were recruited for this research which is part of a micronutrient survey. DBSs were collected onto Whatman 903 filter paper after pricking either middle or ring finger. MMA was quantified in 8‐mm DBS disks using liquid chromatography‐tandem mass spectrometry. DBS and plasma MMA concentrations of a convenience sample of healthy, non‐lactating women (n=182; 19–35 y) residing in Vancouver BC, Canada, served as reference data. A reference value, suggestive of B‐12 deficiency, was computed as 97.5th percentile of DBS MMA concentrations from a subset of the Vancouver convenience sample including women with plasma MMA concentrations < 260 nmol/L (n= 146).

Results

Mean (95% CI) MMA concentration in DBS of Indonesian women (n= 184; aged 27±7 y) was 4.23 (3.90, 4.55) pmol per disk and significantly higher (P= 0.0001; chi‐squared test) than that of Vancouver women [3.18 (2.97, 3.39) pmol per disk; n= 182]. Forty‐three Indonesian women (23 %) had DBS MMA concentrations above the reference value of 5.03 pmol per disk (i.e., the 97.5th percentile). The rate of Vancouver women (9.3 %; n= 182) who had DBS MMA concentrations above the reference value was significantly lower (P< 0.01; chi‐squared test) than the rate of Indonesian women.

Conclusions

DBS MMA has the potential to identify populations with increased risk of low B‐12 status. Lactating women living in rural Indonesia seem to have lower functional B‐12 status than healthy, non‐lactating Vancouver women. Thus, rural Indonesian women and their breastfed infants may be at increased risk for B‐12 deficiency and associated adverse health outcomes.

Support or Funding Information

The study was, in part, funded through Meat & Livestock Australia. YL acknowledges funding from the Canada Research Chair Program of the Canadian Institutes of Health Research and THS acknowledges funding from the University of British Columbia in form of a Four Year Doctoral Fellowship.
A Probabilistic Intake Model to Estimate the Impact of Reformulation by the Food Industry in Irish Consumers
Authors: Pigat Sandrine , Cushen Maeve , O'Mahony Cian , Giammanco Stefania , Connolly Aileen
Published: November 2023

Background

The etiology of the globally growing overweight and obesity epidemic is known to be multi‐causal. The food and beverage industry has made concerted efforts to reformulate their products towards nutritionally healthier options. This project looked at the reformulation efforts of major companies from the food industry and the impact these had on the Irish population.

Methods

Nutrient composition data were collected for approximately 600 products from members of the Food and Drink Industry in Ireland for two time points, 2005 and 2012. Additionally, sales data to assess brand shares at those two time points were obtained from Kantar Worldpanel. To assess the intakes in the population for the two years, the products were matched to the foods consumed in the Irish national dietary consumption surveys from the Irish Universities Nutrition Alliance (IUNA) for adults (18 years+, n=1,500), teenagers (13 to 17 years, n=441), children (5 to 12 years, n=594) and preschool children (1 to 4 years, n=500). Using nutrient composition data provided by participating companies, weighted by market shares, statistical distributions describing the nutrient profiles of food categories of interest were constructed and replaced with the existing data in the IUNA databases for these products. To assess the impact that reformulation of these products had on Irish consumers, probabilistic intake assessments were run for baseline and post‐reformulation time points using the Creme Nutrition(R) model.

Results

The reformulation efforts assessed in this study reduced average daily energy intakes by up to 12kcal/d in adults, 15kcal/d in teens, 10kcal/d in children and 9kcal/d in preschoolers from their total diet. Mean daily fat intakes were reduced between 0.62g/d (preschoolers) and 2g/d (children). In high consumers (P97.5), total dietary sodium intakes decreased by between 520mg/d (preschoolers) and 1,160mg/d (teenagers) and saturated fat intakes were reduced by up to 8g/d among teenagers. Mean daily sugar intakes were reduced by up to 3.5g/d among children. The sugar intakes of teenagers deemed to be high consumers (P97.5) decreased by 8g/d.

Discussion & Conclusion

This model captured the impact that reformulation efforts by industry had on Irish consumers between two time points. However, other factors that were likely to have differed between the two years such as dietary behavior and portion sizes were not captured in this study. Further data on products sold on the market would refine this analysis and create a more complete picture of the overall impact. Daily intakes of energy, sugar, fat, saturated fat and sodium were reduced in all, and significantly (p<0.01) in some cases.

Support or Funding Information

This project was funded by the Food and Drink Industry Ireland and the methodology and design was supported by the Food Safety Authority of Ireland
Evaluating Association between Fast Food Consumption and Use of Internet and Mobile Phone: A Cross Sectional Study by Use of Internet
Authors: Yoshizawa Kazuko
Published: November 2023

Background

Increased fast food consumption has been linked to obesity and overweight. Some studies reported that reasons for fast food use were associated with taste, cost, convenience, energy density, and variety. Use of internet and mobile phone has become very popular, and that might change people's lifestyle including dietary habit. Objective of this study is to evaluate if frequency of use of fast food is associated with increased hours of use of internet and mobile phone.

Method

This study is a cross sectional by use of internet. In December, 2012, a questionnaire was disseminated to women who were living in 47 prefectures in Japan. For analyses in this study 3,192 mothers who had a child or more were included. The subjects had been registered for other panel studies before they participated in this study. Multivariate linear regression analysis was used to evaluate association between fast food use and hours to use internet and mobile phone. Results are statistically significant if a p‐value was less than 0.05. All p‐values are two tailed.

Results

Majority (95%) of the subjects were married while the rest were unmarried or divorced. Ten percent of the subjects lived in Tokyo followed by Osaka (8.5%) and Kanagawa (8.2%). Among the reasons for use of fast food, 28% of mothers chose “because my child likes them” as the primary reason followed by “price is cheaper than other food” (18%). Frequency of fast food use was positively correlated with hours of internet and mobile phone use, and hours of TV watching. It is also positively correlated with the income levels up to the average level and negatively correlated as the levels decrease. Multivariate linear regression analysis was conducted by use of frequency of fast food use as a dependent variable, and hours to use internet and mobile phone, hours of TV watching, income level and age as independent variables. The results showed that age (p<0.000) and hours to use of internet and mobile phone (p<0.019) were statistically significant while hours of TV watching (p<0.827) and income levels (p<0.934) were not important factors.

Conclusion

Fast food use among mothers is associated with increased hours of internet and mobile phone use after adjustment of age, income levels and TV watching: however, further analyses are needed.

Support or Funding Information

2012 Sugiyamajogakuen University Gakuen Research Grant C
F‐actin Assembly And Cytoskeletal Contraction Mediate TNF‐α Induced Hydrogel Retraction, Stiffness, And Nodule Formation By Murine Valvular Cells
Authors: Lim Jina , Ehsanipour Arshia , Lu Jinxiu , Wu Alexander , Pedego Taylor , Walthers Chris , Demer Linda , Seidlits Stephanie , Tintut Yin
Published: November 2023

In calcific aortic valve disease (CAVD), a widespread and rapidly fatal disorder, the valve cusps undergo retraction, stiffening, and nodular calcification. Therapeutic options are limited to invasive procedures. The pro‐inflammatory cytokine, tumor necrosis factor‐alpha (TNF‐α), has been shown to contribute to CAVD progression; however, the mechanisms of its actions on cusp retraction and stiffening are not clear.

We investigated the effects of TNF‐α on murine aortic valvular interstitial cells (VIC) within a 3‐dimensional, free‐floating, compliant collagen hydrogel to simulate their natural substrate. Results showed that TNF‐α increased retraction (percent diameter), stiffness (Pascal), and formation of calcified nodular structures (Alizarin Red positivity) in the VIC‐laden hydrogels. Effects of TNF‐α on retraction, stiffness, and nodule formation were attenuated by inhibition of myosin II‐mediated cytoskeletal contraction with blebbistatin. Inhibition of actin polymerization with cytochalasin D, but not inhibition of Rho‐kinase with Y27632, blocked TNF‐α‐induced retraction in 3‐D VIC hydrogels, suggesting that actin stress fibers mediate TNF‐α‐induced effects. In the hydrogels, TNF‐α increased levels of phospho‐IκB, and co‐treatment with an NF‐κB inhibitor blocked TNF‐α‐induced retraction and stiffening, suggesting that NF‐kB signaling mediates the effects of TNF‐α. TNF‐α also significantly increased collagen deposition, as visualized by Masson's trichrome staining, and it upregulated expression of discoidin domain‐containing receptor 2, fibronectin, and smooth muscle alpha‐actin, as evidenced by realtime RT‐qPCR. In human aortic valves, we found that stiffness and collagen deposition were greater in calcified than non‐diseased cusps.

In conclusion, inflammation may induce valvular cusp retraction, stiffening, and nodule formation by promoting F‐actin assembly and cytoskeletal contraction.

Support or Funding Information

This work was supported by funding from the National Institutes of Health (DK081346‐S1, HL114709, HL121019). J.L was supported by UCLA Children's Discovery and Innovation Institute Harry Winston Fellowship Award. S.S. and A.E. were supported by the University of California, Los Angeles Henry Samueli School of Engineering and Applied Sciences.
Histomorphometry of Intra‐Acinar Vessel Muscularization in the Rat Sugen/Hypoxia Efficacy Model of Pulmonary Arterial Hypertension
Authors: Syed Jameel , Brenneman Karrie , Zhan Yutian , Nguyen Vuong , Percival Karen , Schlerman Frank , Yang Zhiyong , Evans Steven , O'Neil Shawn
Published: November 2023

Pulmonary arterial hypertension (PAH) is a debilitating disease in which increased cell proliferation in the walls of small arteries in the lung causes abnormal vessel wall thickening and luminal narrowing resulting in increased arterial pressure and secondary right‐sided heart failure. Vasodilators (eg, Sildenafil [Revatio®]) are the current standard of care and may extend patient survival; however, no disease‐modifying therapeutics are currently available. In an effort to meet this unmet medical need, we have evaluated key morphometric endpoints in lung samples from the rat sugen/hypoxia (Su/Hx) efficacy model of PAH to confirm successful disease induction and quantify the increase in muscularization of intra‐acinar vessels that characterizes PAH. A modified elastin stain and dual immunohistochemical methods (αSMA/Ki67 and CD31/phosphoRb) were developed to quantify the wall thickness of intra‐acinar vessels (≤ 50 um external diameter) (elastin), the density of fully muscularized intra‐acinar vessels per mm2 (αSMA), and number of proliferating cells per mm2 (Ki67+ and pRb+) associated with vascular remodeling. For quantification, we used Visiopharm newCast stereology software and applied a random‐field sampling method with observer‐assisted identification of vasculature and quantification. For these respective endpoints, means for Su/Hx groups were 1.74×, 2.26×, 4.36×, and 28.00× higher compared with normoxic control groups, confirming successful disease induction, which correlated well with terminal hemodynamic outcomes. These histomorphometric tools are now well positioned to support future efficacy model evaluations.
Immunohistochemical Evaluation of Five Antibodies for the Detection of Simian Immunodeficiency Virus
Authors: Machiah Deepa , Villinger Francois , Gumber Sanjeev
Published: November 2023

Similar to humans with HIV who develop encephalitis, rhesus macaques infected with Simian Immunodeficiency Virus (SIV)are susceptible to develop encephalitis characterized by perivascular accumulation of virus‐laden macrophages and multinucleated giant cells, astrogliosis, microgliosis, and neuronal injury. There are no investigative studies in the literature documenting sensitivity and specificity of available Anti‐SIV antibodies for the detection of viral protein in the tissue samples and therefore, a more formal and comprehensive investigation is warranted to advance our understanding of the disease. In this study, a comparative immunohistochemical analysis was carried out on formalin‐fixed, paraffin embedded sections of lymphoidorgans and brain from SIV infected rhesus macaques (Macaca mulatta), experimentally infected with SIVmac251 or SIVmac239. Immunohistochemistry (IHC) was performed using five different antibodies: Anti‐SIV mac251gag mAb (clone FA2), Anti‐SIV mac251 p28 mAb (HRP, clone 3F7), Anti‐SIV mac251 p28 polyclonal antibody, Anti‐SIV mac251 gp130 mAb(clone 5i45) and Anti‐SIV mac251 p17 gag mAb (clone KK59) to determine their efficacy for the detection of SIV viral protein. Immunohistochemical quantitative data are correlated with the viral load to estimate sensitivity of each antibody.
Effects of Resveratrol in CXCR4 Expression and Apoptotic Markers in Normal Cell Lines Derived from a Breast Cancer Patient
Authors: Arroyo Gerardo Antonio , Figueroa Maria , Muñoz‐Forti Kevin , Maldonado Alexandra , Torres Bryan , Bernard Faviola , Almodovar Sharilyn , Suarez Edu , Ruiz Abigail
Published: November 2023

The cellular and molecular environment of normal cells from breast cancer patients are particulary understood. Chemokines and their receptors have roles in cellular proliferation, metastasis, and angiogenesis. Specifically, CXCR4, a G‐protein coupled receptor is expressed in high levels in breast neoplastic tissues and in low levels in normal breast tissue. CXCR4 expression may induce proliferation, migration, invasion and angiogenesis. Recent studies have identify resveratrol (a natural polyphenol) to play a role on inflammation, apoptosis, and neovascularization. Due to limited scientific literature on the effects of resveratrol on CXCR4, the biological process in which both coincide, (inflammation, apoptosis, and neovascularization), and the potential use of resveratrol, as a therapeutic agent in cancer, we studied the effect of this polyphenol on CXCR4. In addition, we explored the effects on the apoptosis mediator Bax. We hypothesized that resveratrol increases the expression of CXCR4 in normal cells from a breast cancer patient. We determined the effects of resveratrol at concentrations of 25, 50, 100, 200 μM after 24 hours of treatment in the protein levels of CXCR4 and Bax‐2 by western blot using CCD1097 a fibroblastic cell line, derived from normal skin from the breast of a patient undergoing biopsy of a grade III infiltrating ductal carcinoma. Our preliminary results indicate that higher doses increased expression of CXCR4 in a dose‐dependent compared to control vehicle treated cells. Similarly, Bax expression increased in all treated cells but in a non‐dependent dose manner. These results may suggest that resveratrol have an effect on CXCR4 protein levels, leading to an increase in proliferation in normal cells from cancer patients. As reported, at lower doses resveratrol may have antiapoptotic effects and at higher doses may induce apoptosis. Resveratrol may prompt for clearance of cancer cells using different pathways but additional experiments will be necessary to identify the exact mechanism of action. Additional experiments are underway to identify the proliferative effects of resveratrol including apoptosis (caspases) and inflammation (TNF‐α and SOD‐2).

Support or Funding Information

This work was supported by UPR PRISE Program NIH Grant #R25GM096955.
Discovery of Potent GOAT inhibitors to Elucidate the In Vivo and In Vitro Mechanism Of Action of GOAT
Authors: Yang HsiuChiung , Chen Yanyun , Reynolds Nichole A , Mosior Marian K , Brier Richard A , Hembre Erik J
Published: November 2023

Ghrelin (AG) is the only known orexigenic peptide in human and rodents. It is converted from its precursor, unacylated ghrelin (UAG), to the biologically active peptide, acylghrelin. Two independent groups (Yang et al., 2008; Gutierrez et al 2009) using different approaches identified the same protein, Ghrelin O‐Acyl Transferase (GOAT), as the long sought enzyme which is responsible for the bioactivation of ghrelin peptide. Results from GOAT gene knockout mice confirmed that GOAT is the principal enzyme responsible for the acylation of ghrelin. ( Zhao et al 2010)

To enable the discovery of GOAT inhibitors, we have developed GOAT enzymatic and cellular assays. Using these assays, we have discovered potent GOAT inhibitors. In rodent models, these inhibitors exhibited similar effects on the acylated ghrelin and unacylated ghrelin to that observed in the GOAT gene knockout mice. In addition, we utilized GOAT inhibitors to elucidate the detailed molecular mechanism of the GOAT enzymatic reaction, which involves two substrates, c‐CoA. These pharmacological agents also allowed us to build an in vivo ghrelin processing model. The ghrelin processing model provided us with important insights to answer key questions regarding ghrelin metabolism and regulation in vivo.
Combretastatin augments the anticancer effect of vincristine either in vivo or in vitro
Authors: El‐Moselhy Mohamed Ahmed , Aboubakr Esam Mohamed , Ali Omar Mohamed , Taye Ashraf
Published: November 2023

Hepatocellular carcinoma (HCC) is a serious healthcare problem worldwide especially in the Middle East because of its increasing morbidity and mortality.

Aim

The present study was conducted both in vivo and in vitro to investigate the anti‐cancer effect of combretastatin either alone or in combination with vincristine and the mechanisms underlying this effect.

Methods

In vivo study: a total 50 Wister albino rats were divided into five groups 10 rats for each, first group was i.p injected with normal saline and used as a normal groups. The other four group were injected i.p with 200mg/kg DENA. Two weeks later animals were orally administered 3ml/kg CCl4 once a week for 8 successive weeks. At the end of 8th week animals were divided into the following groups: Normal group, control group (hepatocellular carcinoma induced), combretastatin (200mg/kg) i.p treated group, combretastatin (100 mg/kg) i.p + vincristine (25 mg/kg) i.p treated group and vincristine (50 mg/kg) i.p treated group. The following parameters were assessed GSH, MDA, AFP, ALT, AST, tissue carbonyl content, number of hepatic nodules, histopathological examination of the hepatic tissue, tubulin inhibition in the hepatic tissue and the relative hepatic weight. In vitro study human hepatocarcinoma cell line (HepG2) were used to evaluate the cytotoxic and tubulin protein inhibitory effects of the tested drugs.

Results

The results of the in vivo study revealed that combretastatin, combretastatin + vincristine and vincristine groups showed a significant decrease in GSH, AFP, number of hepatic nodules, ALT, AST and tubulin concentration in the hepatic tissue compare to the control group.

Moreover the co‐administration of combretastatin with vincristine showed a significant increment in these parameters compared to single administration of combretastatin or vincristine, also MDA concentration and tissue carbonyl content concentrations were significantly increased compared to control group. On the other hand tubulin concentration and the relative hepatic weight were significantly less than control group. Also the results of the in vitro study revealed that both combretastatin and vincristine showed significant cytotoxic and tubulin inhibitory effects, while these effects were significantly enhanced by simultaneous administration of both drugs.

Conclusion

combretastatin showed a promising anticancer activity against hepatocellular carcinoma by three mechanisms; tubulin inhibition leading to hepatic tumor blood supply shutdown, decrease reduced glutathione concentrations and oxidative stress generation. Furthermore these effects were greatly enhanced by co‐administration with vincristine.

Moreover the present study suggests that we can greatly enhance the anti‐cancer activity of narrow therapeutic windows drugs such as vincristine by co‐administration with combretatstatin leading to fewer side effects.
Investigation of the regulatory effects of saccharin on cytochrome P450s in male ICR mice
Authors: Jo Jun Hyeon , Kim Sun Joo , Lee Sangkyu
Published: November 2023

Saccharin, the first artificial sweetener, was discovered in 1879. Saccharin does not have any calories and is approximately 200–700 times sweeter than sugar. Saccharin was the most common domestically produced sweetener in Korea in 2010, and it has been used as an alternative to sugar in many products. The interaction between artificial sweeteners and drugs may affect the drug metabolism in patients with diabetes, cancer, and liver damage, this interaction has not been clarified thus far. Here, we examined the effects of the potential saccharin‐drug interaction on the activities of 5 cytochrome P450 (CYPs) in male ICR mice; further, we examined the effects of saccharin (4,000 mg/kg) on the pharmacokinetics of bupropion after pretreatment of mice with saccharin for 7 days and after concomitant administration of bupropion and saccharin. Our results showed that compared to control, saccharin did not have a significant effect on the 5 CYPs in the S9 fractions obtained from the liver of male ICR mice. In addition, we observed no differences in the pharmacokinetic parameters of bupropion between the control group and the groups pretreated with saccharin and that receiving concomitant administration of saccharin. Thus, our results showed that saccharin is safe and the risk of saccharin‐drug interaction is very low.

Support or Funding Information

The National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Science, ICT and Future Planning) (No. NRF‐2014M3A9D9069714).
Lactucopicrin suppresses oxidative stress initiated by scopolamine‐induced neurotoxicity through activation of NRF2 pathway
Authors: Subedi Lalita , Venkatesan Ramu , Park Yong Un , Kim Sun Yeou
Published: November 2023

The cholinergic antagonist scopolamine induced dementia with impairment of memory and cognitive functions, as seen in Alzheimer's disease (AD). This scopolamine‐induced AD model has been used worldwide to understand, identify, and characterize beneficial therapeutic targets. The present study initiate to investigate the molecular mechanisms underlying scopolamine‐induced cholinergic neuronal dysfunction and the neuroprotective effect of lactucopicrin, an inhibitor of acetylcholine esterase. We focused on apoptotic cell death, caspase activation, reactive oxygen species (ROS) generation, mitochondrial dysfunction, and the expressions of anti‐ and pro‐apoptotic proteins in C6 cells. We also analyzed the expression of antioxidant enzymes and their transcription factor nuclear factor (erythroid‐derived 2)‐like 2 (NRF2) in C6 cells, along with neurite outgrowth in N2a neuroblastoma cells. Our results revealed that scopolamine pre‐treated for 1 h induce cytotoxicity by increasing apoptotic cell death via oxidative stress‐mediated caspase 3 activation and mitochondrial dysfunction. Scopolamine also downregulated the expression of antioxidant enzymes, such as superoxide dismutase, glutathione peroxidase, and catalase, and NRF2 expression. Lactucopicrin treatment protected C6 cells from the scopolamine‐induced toxicity by reversing the effects on these toxicity markers. In addition, scopolamine attenuated the secretion of neurotrophic nerve growth factor (NGF) in C6 cells and neurite outgrowth in N2a cells. As expected, lactucopicrin enhanced NGF secretion and neurite outgrowth in these neuronal cells. Our study suggests that first report about lactucopicrin, a potential neuroprotective candidate, ameliorating scopolamine‐induced cholinergic dysfunction via NRF2 activation and subsequent expression of antioxidant enzymes

Support or Funding Information

This research was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean government, MSIP (NRF‐2014M3A9B6069338)
Stapled B chain analogues of relaxin‐3 retain biological activity
Authors: Dawe Gavin Stewart , Jayakody Tharindunee , Mawari Subhi
Published: November 2023

Relaxin‐3, the most recently discovered member of the relaxin/insulin‐like family of peptides, is involved in regulating stress and anxiety, feeding, metabolism and cognition. Mature relaxin‐3 consists of two peptide chains (A and B) held together by disulphide bonds. Structure activity relationship studies have revealed that the relaxin‐3 B chain is more important in binding and activating the cognate receptor of relaxin‐3, relaxin/insulin‐like family peptide receptor 3 (RXFP3), than the A chain. Stapling renders peptide drugs less prone to proteolysis and can increase activity by locking the peptide conformation. We synthesized stapled B chains of relaxin‐3, hypothesizing that stapling mimics the function of the A chain in stabilizing the B chain and does not compromise activation of RXFP3. Relaxin‐3 B chain was stapled at the 14th and 18th positions or the 18th and 22nd positions by i and i+4 ring‐closing metathesis hydrocarbon stapling. In order to determine RXFP3 activation, we tested the peptides for cAMP inhibition and ERK1/2 activation. Both peptides significantly activated RXFP3 as observed in the inhibition of forskolin induced cAMP assay and ERK1/2 activation assay. We conclude that stapling of the relaxin‐3 B chain does not prevent its ability to activate RXFP3. Stapling of the relaxin‐3 B chain holds promise for development of stable peptide agonists and antagonists of RXFP3 to aid relaxin‐3/RXFP3 research.

Support or Funding Information

This work was supported by the Biomedical Research Council of Singapore (BMRC 10/1/21/19/645), the National Medical Research Council of Singapore (NMRC/1287/2011) and the Ministry of Education, Singapore, Academic Research Fund Tier 1 Seed Fund for Basic Science Research (T1‐BSRG 2014‐03).
G protein‐coupled Receptor 30 (GPR30) PDZ‐dependently and Constitutively Increases ERK1/2 Signaling Through Calcineurin and Kinase Suppressor of Ras 2 (KSR2)
Authors: Olde Bjorn , de Valdivia Ernesto Gonzalez , Broselid Stefan , Kahn Robin , Leeb‐Lundberg Lars M. F.
Published: November 2023

The objective of the present study was to map the mechanism whereby G protein‐coupled receptor 30 (GPR30), also called G protein‐coupled estrogen receptor (GPER), stimulates extracellular signal‐regulated kinase (ERK)1/2 signaling. GPR30 plays important roles in cancer and cardiometabolic regulation. We showed recently that GPR30 forms a plasma membrane complex through its C‐terminal type I PSD‐95/Discs‐large/ZO‐1 homology (PDZ) motif with a membrane‐associated guanylate kinase (MAGUK) and protein kinase A (PKA)‐anchoring protein 5 (AKAP5), and AKAP5‐anchored PKA regulatory subunit RII suppresses receptor endocytosis and enables the receptor to constitutively inhibit cAMP production. Here, we investigated if this PDZ‐dependent GPR30 complex also regulates ERK1/2 signaling. To do so, human and mouse GPR30 were ectopically expressed in HEK293 cells and MDCK cells, and receptors and effectors were monitored by immunoblotting, immunoprecipitation, confocal immunofluorescence microscopy, split luciferase reporter techniques, and reporter gene assays. We found that GPR30 constitutively increased extracellular signal‐regulated kinase (ERK) 1/2 activity in several cell systems. The response was dependent on an intact receptor PDZ motif. Furthermore, knocking down AKAP5 or inhibiting calcineurin with FK506 inhibited the receptor response. GPR30 PDZ‐dependently inhibited basal NFAT signaling, consistent with the receptor favoring AKAP5‐anchoring of calcineurin. The calcineurin substrate kinase suppressor of Ras 2 (KSR2), a mitogen‐activated protein kinase (MAPK) scaffold, enhanced the GPR30‐promoted response, also dependently on the receptor PDZ motif. GPR30 also PDZ‐dependently favored a membrane KSR2 complex at the expense of the monomeric form. On the other hand, disrupting AKAP5‐PKA RII interaction with St‐Ht31, or inhibiting protein kinase C (PKC) with GF109203X or epidermal growth factor receptor (EGFR) tyrosine kinase with AG1478 had no effect on the GPR30‐stimulated response. FK506 also increased the amount of GPR30 in the plasma membrane, thus acting opposite to St‐Ht31, which increased receptor endocytosis. We conclude that the PDZ‐dependent GPR30 complex with AKAP5 includes calcineurin, which favors GPR30 endocytosis and enables the receptor to constitutively increase ERK1/2 signaling through KSR2.

Support or Funding Information

Swedish Cancer Foundation
Osteomeles schwerinae extracts inhibits the binding to receptors of advanced glycation end products and TGF‐β1 expression in mesangial cells under diabetic conditions
Authors: Kim Young Sook , Jung Dong Ho , Lee Ik‐Soo , Pyun Bo‐Jeong , Kim Jin Sook
Published: November 2023

Background

Osteomeles schwerinae C. K. Schneid. (Rosaceae, OSSC) is a medicinal plant traditionally used to treat various diseases in Asia. The chemical constituents of OSSC have an inhibitory effect on aldose reductase activity, which has been implicated in the pathogenesis of diabetic complications. However, the protective effects of the pharmacological activity and potential mechanisms in diabetic nephropathy are still not known.

Objective

In the present study we investigated that OSSC extracts and major compounds were examined for their effects on binding to the receptors of advanced glycation end products (RAGE) and on transforming growth factor‐beta1 (TGF‐β1) expression‐related signal mechanisms in mouse glomerular mesangial cells (GMCs).

Materials and methods

A simple, rapid and efficient method was developed for the simultaneous determination of the marker compounds in the ethanol extract of the leaves and twigs of OSSC using HPLC‐diode array detector (DAD). In this study, we determined the effects of OSSC extract and hyperoside on AGE and RAGE binding, and studied the mechanism of OSSC extract effects on AGE‐bovine serum albumin (BSA)‐treated GMCs. GMCs overexpressing human RAGE were cultured in AGE‐BSA labelled with Alexa 488, and OSSC extract. AGE/RAGE binding were measured using fluorescence (excitation 485 nm/emission 528 nm). TGF‐β1 protein expression levels were determined by western blot analyses.

Results

OSSC extracts of leaves and twigs (1 mg/ml) inhibited on AGE/RAGE binding and TGF‐β1 protein expression in a dose‐dependent manner in GMCs. Furthermore, OSSC extracts reduced the effects on AGE‐BSA‐induced reactive oxidative species (ROS) formation and nuclear translocalization of transcription factor NF‐κB. OSSC extracts inhibited phosphorylation of extracellular signal‐regulated protein kinases1/2 (ERK1/2), p38 mitogen‐activated protein kinases (p38MAPK), and IκB. Hyperoside also inhibited AGE/RAGE binding and ROS formation, and reduced TGF‐β1 expression and IkB phosphorylation.

Conclusions

OSSC extracts and hyperoside may attenuate AGE/RAGE binding and expression of TGF‐β1 by downregulating of pERK1/2, p38MAPK and IκB phosphorylations in GMCs under diabetic condition and retard the development of diabetic complications such as diabetic nephropathy.

Support or Funding Information

This research was supported by Grants (K14040 and K15270) from the Korea Institute of Oriental Medicine (KIOM).
New Triterpenoid Compounds stimulates Glucose Uptake Through IRS‐1‐Akt and Erk1/2‐GLUT4 Pathways in C2C12 myotubes
Authors: Han Joo‐Hui , Zhou Wei , Na MinKyun , Myung Chang‐Seon
Published: November 2023

Regulation of blood glucose levels within normal range is important for treating diabetes mellitus and one of the strategies is improving glucose uptake into insulin‐sensitive tissues, including skeletal muscle and adipose tissues. The present study has investigated the effect of new triterpenoid compounds on glucose uptake in C2C12 myotubes and in clarifying the underlying molecular mechanism, focusing on glucose uptake related insulin pathways. To evaluate the activity of triterpenoid compounds 1 and 2, we examined glucose uptake ability and GLUT4 translocation in C2C12 myotubes. Compounds 1 and 2 significantly increased basal‐and insulin‐stimulated glucose uptake and GLUT4 translocation from cytoplasm to plasma membrane in C2C12 myotubes. Both compounds stimulated the insulin‐dependent pathway, insulin receptor substrate (IRS)‐1, protein kinase B (Akt), glycogen synthase kinase (GSK)‐3β and extracellular signal‐regulated kinase (Erk) 1/2 phosphorylation. But insulin‐independent pathway, which is 5′ adenosine monophosphate‐activated protein kinase (AMPK), acetyl‐CoA carboxylase (ACC) was not affected. Inhibition of signal activation using Akt inhibitor, triciribine, or Erk1/2, U0126 decreased the ability of both compounds to enhance basal‐ and insulin‐stimulated glucose uptake. These results indicate that compounds 1 and 2 activate both IRS‐1/Akt and Erk1/2 pathways and subsequently stimulate GLUT4 translocation, leading to increase glucose uptake activity in C2C12 myotubes. These findings supports the use of triterpenoid compounds in the regulation of blood glucose level and increase insulin sensitivity.