bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of phosphodefective FFAR4 C‐terminal mutants on COX‐2 expression in macrophages
Authors: Cheshmehkani Ameneh , Moniri Nader H. 
Published: November 2023

Macrophages have critical roles in the initiating, maintaining, and resolving inflammation, and mediate this response partially through expression of cyclooxygenase‐2 (COX‐2), which generates prostaglandins (PG). Free‐fatty acid receptor‐4 (FFAR4), a G protein‐coupled receptor that was previously termed GPR120, has been shown to play important roles in regulating macrophage function to produce robust anti‐inflammatory effects. In this regard, agonism of FFAR4 with docosahexaenoic acid (DHA) has previously been shown to reduce expression of COX‐2 and subsequent PGE2 synthesis. Our previous work, as well as that of others, has shown that FFAR4 signaling is regulated by phosphorylation of the receptor on its C‐terminal tail and that mutation of these phosphorylation sites diminishes the receptors ability to interact with β‐arrestin partner proteins. The goal of the current study was to assess the role of the C‐terminal tail of FFAR4 in regulating COX2 expression and PGE2 synthesis. Here, we have created C‐terminal truncated mutants of both the short (FFAR4‐S) and long (FFAR4‐L) isoforms of FFAR4 and stably expressed them in murine RAW264.7 macrophages. Our results show that stable expression of wild‐type (WT) FFAR4‐S in RAW cells significantly reduces COX2 expression, while expression of the C‐terminal truncated mutant of FFAR4‐S does not affect COX2 expression compared to control RAW macrophages. Meanwhile, stable expression of WT‐FFAR4‐L, which has previously been shown to be intrinsically biased solely towards β‐arrestin signaling, also resulted in reduced COX‐2 expression, while expression of the truncated FFAR4‐L mutant reversed this effect. These results suggest that C‐terminal phosphorylation and subsequent β‐arrestin recruitment are integral in FFAR4‐mediated alteration of COX2 expression in macrophages.Support or Funding InformationSupported by NIH/NIDDK grant DK098730 to N.H.M.
Apelin‐13 inhibits lipid accumulation in human hepatocytes
Authors: Park Soojin , Huang Jin , Im Dongsoon 
Published: November 2023

BackgroundApelin is an adipokine and acts through a G protein‐coupled receptor named APJ. Its protective function on lipid accumulation in adipocytes has been proven. However, functions of apelin in hepatocytes have not been investigated.METHODSIn order to assess functional role of apelin and APJ in hepatocytes, we used an in vitro model of liver X receptor‐mediated hepatocellular steatosis. In human hepatoma cells, APJ expression was confirmed.RESULTSA specific liver X receptor activator induced lipid accumulation and apelin‐13 inhibited the lipid accumulation. Apelin‐13‐induced inhibition was ameliorated by treatment of F13A, a specific APJ antagonist. Furthermore, silencing of APJ by siRNA transfection also inhibited apelin‐13 action. Liver X receptor activator induced expression of SREBP‐1c, a key transcription factor for lipogenesis. Apelin‐13 inhibited the SREBP‐1c induction and the inhibition was ameliorated by F13A in the hepatocytes. Treatment of pertussis toxin, LY294002, U0126, and SB202190 blocked apelin‐13‐induced inhibition of lipid accumulation, suggesting involvement of Gi/o proteins, PI‐3‐kinase, and MAPK kinases.CONCLUSIONTherefore, the present data suggest that apelin may function to protect lipid accumulation in the liver.
Effects of Pentadecapeptide BPC 157 on Experimental Rat Model of Dry Eye
Authors: Kralj Tamara , Kokot Antonio , Kasnik Katarina , Drmic Domagoj , Zlatar Mirna , Seiwerth Sven , Sikiric Predrag 
Published: November 2023

AimWe hypothesize the stable gastric pentadecapeptide BPC 157 as a therapy of dry eye syndrome in rats. BPC 157 already cures ocular tissues. BPC 157 eye drops (ug‐ or ng‐regimen) rescued total debridement of corneal epithelium (Coll Antropol 2005;29(1):321–5), successfully closed perforating corneal incisions in rats (Exp Eye Res 2015;136:9–15), counteracted particular NO‐specific parallel miotic effect of L‐NAME and L‐arginine and atropine‐induced mydriasis in rats and guinea pigs (Eur J Pharmacol 2016;771:211–9). In addition, BPC 157 counteracts various ulcerative lesions in skin and gastrointestinal tract, and BPC 157/NO‐relationship is established in various experimental models and species (Curr Pharm Des 2014;20(7):1126–35), providing that it would counteract the damaging effects of lacrimal gland extirpation, and dry eye syndrome. Dry eye syndrome is a relatively common disease with multifactorial causes, which may cause simptoms variating from discomfort to blindness. This study aimed to evaluate the effect of BPC 157 on the rat dry eye model.MethodsWe used male Albino Wistar rats, 200g, randomly divided. After application of Ketmine‐HCl 50–60 mg/kg + Xylazine‐HCl 5–10 mg/kg, Tetrakain 0,5% drops, left extraorbital lacrimal gland was isolated from surrounding tissues and surgically removed. Rats received stable gastric pentadecapeptide BPC 157 (Diagen, Slovenia) topically (5 drops (2 μg/ml)) or an equivolume of destilled water (controls). Corneal defects were analyzed with microcamera observation using blue light after 0.5% fluorescein eye drop instillation. These examinations were performed in both treated and control eyes at baseline and on weeks 1, 2 and 4.ResultsPentadecapeptide BPC 157 administered topically significantly reduced clinical signs of dry eye by decreasing corneal defect in all treated groups (Figure 1), while in control groups corneal defect persisted (Figure 2)ConclusionBPC 157 was originally an anti‐ulcer peptide used in trials for ulcerative colitis and now is in trials for the treatment of multiple sclerosis (Curr Pharm Des 2014;20(7):1126–35). Now, we demonstrated that pentadecapeptide BPC 157 counteracts dry eye lesions in rats.Support or Funding InformationUniverity of Zagreb, Zagreb, Croatia (Grant number BM099)
Anti‐inflammatory Evaluation Of LASSBio‐1823, A New p38 MAPK Inhibitor
Authors: de Carvalho Patrícia Ribeiro , de Morais Cordeiro Natália , Freitas Rosana Ribeiro , Alves Marina , Manssour Carlos Alberto Ribeiro , Fernandes Patricia Ribeiro 
Published: November 2023

Protein kinases are directly involved in many human disorders, including inflammatory diseases and cancer. The p38 mitogen‐activated protein kinase (MAPK) plays a crucial role in a signal transduction cascade regulating biosynthesis and release of pro‐inflammatory cytokines. Our aim was to evaluate LASSBio‐1823 effects using in vitro and in vivo models of inflammation.LASSBio‐1823 was evaluated in vitro p38 MAPK assay, carrageenan‐induced leukocyte migration into subcutaneous air pouch (SAP) of mice and measurement of nitric oxide (NO) and TNF‐α obtained from SAP exudates and after activation of J774.A1 macrophage cell line in vitro. J774.A1 cells were incubated with LASSBio‐1823 (1, 10 or 30 μM) and further activated with lipopolysaccharide (LPS, 1 μg/mL). After 24 h the supernatant was collected and NO and TNF‐α measured. Mice (Swiss Webster, 22–25g, n=4–6, authorization of use #DFBCICB015‐04/16 (COBEA/UFRJ/Brazil)) were orally treated with LASSBio‐1823 (10 μmol/kg) 1 h before carrageenan injection in SAP. After 24 h animals were euthanized and exudate was collected for subsequent dosages. Results are presented as mean±SD. Statistical analysis was ANOVA and Bonferroni post‐test (*p<0.05).Results obtained in p38 enzymatic assay demonstrated LASSBio‐1823 IC50 of 21.33 μM (10.5–43.5 μM). The compound reduced NO production by LPS‐activated J774.A1 cells: 1 μM: 77.5±27 μM, 10 μM: 7.5±5.2 μM* and 30 μM: 14.6±11.2 μM* when compared to LPS: 85.5±20.1 μM. It was also observed a reduction in TNF‐α levels in LPS‐activated cells: 1 μM: 1,430.3±48.5 pg/mL, 10 μM: 865.6±53.9 pg/mL* and 30 μM: 324.0±27.2 pg/mL* when compared to LPS: 1,313.8±146.0 pg/mL. LASSBio‐1823 (10 μmol/kg, p.o.) also reduced leukocyte migration induced by carrageenan (54.3±33.0 cells × 106/mL*) when comparing with carrageenan‐injected group (128±9.9 cells × 106/mL). Levels of NO (91.7±43.0 μM*), TNF‐α (222.4±57.3 pg/mL*) and protein extravasased (69.6±21.8 pg/mL*) were reduced when compared to carrageenan‐treated groups (242.5±117.6 μM, 454.2±148.6 pg/mL and 273.0±14.7 pg/mL, to NO, TNF‐α and protein, respectively).Our results suggest that LASSBio‐1823 reduces some parameters of an inflammatory process thus suggesting as a prototype of a new anti‐inflammatory drug.Support or Funding InformationFinancial support: CAPES, CNPq, FAPERJ and Instituto Vital Brazil (donation of animals).Technical support: Alan Minho.
Tetrachloroethene‐mediated autoimmunity: Significance of lipid‐derived aldehydes
Authors: Wang Gangduo , Wang Jianling , Ansari G.A. Shakeel , Khan M. Firoze 
Published: November 2023

Tetrachloroethene (perchloroethylene, PCE), a ubiquitous environmental contaminant, has been implicated in inducing autoimmunity/autoimmune diseases (ADs), including systemic lupus erythematosus (SLE) and scleroderma in humans. However, experimental evidence suggesting the potential of PCE in mediating autoimmunity is lacking. This study was, therefore, designed to explore PCE's potential in inducing an autoimmune response. Six‐week old female MRL+/+ mice, in groups of 6 each, were treated with PCE (0.25, 0.5 or 1.0 mg/ml) via drinking water for 12, 18 and 24 weeks and markers of lipid peroxidation (oxidative stress) and autoimmunity were determined. PCE exposure led to significant increases in malondialdehyde (MDA)‐protein adducts, and anti‐ MDA‐/4‐hydroxynonenal (HNE)‐protein adduct antibodies, which showed a time‐dependent response. The increases in anti‐MDA‐/HNE‐protein adduct antibodies were associated with significant increases in serum anti‐nuclear antibodies (ANA) and anti‐dsDNA antibodies at 18 weeks and, to a greater extent at 24 weeks, suggesting that PCE exposure exacerbated autoimmunity in our animal model. The splenocytes isolated from mice treated with PCE for 18 and 24 weeks showed greater Th17 cell proliferation and increased release of IL‐17 in culture supernatants following stimulation with MDA‐mouse serum albumin adducts, suggesting that MDA‐modified proteins may act as an immunologic trigger by activating Th17 cells and contributing to PCE‐mediated autoimmunity. Our studies provide the experimental evidence that PCE induces/exacerbates an autoimmune response and lipid‐derived reactive aldehydes (MDA and HNE) contribute to this response.Support or Funding InformationSupported by NIH ES023106.
Let‐7c is the Most Effective among a Set of Bioengineered miRNA Agents in Inhibiting Hepatocellular Carcinoma Cell Proliferation
Authors: Jilek Joseph Lionel , Ho Pui Yan , Duan Zhijian , Yu Ai‐Ming 
Published: November 2023

Hepatocellular carcinoma (HCC) is the most common liver cancer in the world. Few curative drugs are currently on the market, including sorafenib, although efficacy is limited and resistance and recurrence remains a significant problem. Dysregulation of microRNAs (miRNA), a family of short non‐coding RNAs, has been revealed as a significant contributor to HCC signaling, thus posing as a pharmacological target. However, current research is limited to the use of synthetic miRNA reagents that carry extensive artificial modifications. Recently our group has established a robust, fermentation based approach for the production of miRNA agents using a tRNA/pre‐miRNA scaffold. Using this approach, we have generated a small set of biologic miRNA/siRNA agents. Upon transfection, several agents were identified to significantly reduce Huh7 and Sk‐Hep‐1 cell viability. Further study on this subset of hits revealed chimeric let‐7c as the most potent and effective miRNA in inhibiting proliferation of both Huh7 and Sk‐Hep‐1 cell lines, with EC50 values of 2 and 7 nM, respectively. Subsequently, we demonstrated that biologic let‐7c significantly enhanced the sensitivities of Huh7 and Sk‐Hep‐1 cells to doxorubicin and sorafenib. These were accompanied by an 80‐fold and 20‐fold increase in mature let‐7c in Huh7 and Sk‐Hep‐1 cell lines, respectively, as well as suppression of let‐7c target gene expression (e.g., Bcl‐2/Bcl‐xL), following the transfection with biologic let‐7c. Overall, these data demonstrate that biologic let‐7c is a potent antiproliferative agent for HCC, and it also sensitizes HCC cells to sorafenib and doxorubicin.Support or Funding InformationThis study was supported by grant [R01GM133888] from NIGMS, NIH. J.L.J. was supported by a NIGMS Pharmacology Training Program [T32GM099608].
The Anti‐cancer Effects of Resveratrol on Canine Hemangiosarcoma Cells
Authors: Alderete Karen Stefania , Carlson Ariel , Grant Marianne , Sharkey Leslie , Zordoky Beshay 
Published: November 2023

Hemangiosarcoma (HSA) is a highly malignant tumor of vascular endothelial origin that exhibits aggressive biological behavior. HSAs are more common in dogs than other species including cats, sheep, horses, and cattle. Generally, canine HSAs have poor prognosis. After the complete surgical excision of the primary tumor, the survival time of dogs with HSA is typically short, less than 3 months. Adjuvant treatment with chemotherapy, especially anthracycline‐based regimens such as doxorubicin, can result in modest improvement of survival rates. However, the median survival time of dogs with HSA is still short, even after chemotherapy and surgery. Therefore, there is an urgent need to improve the adjuvant chemotherapeutic regimens in order to reach better clinical outcomes in dogs with HSA. Metformin and resveratrol have been shown to have strong anti‐proliferative and/or pro‐apoptotic properties in several human cancer cell lines. However, the potential anticancer effects of metformin and resveratrol have not been determined in canine HSAs. Therefore, the objective of this study is to investigate the potential growth inhibitory effects of metformin and resveratrol on HSA cell lines when used alone or in combination with doxorubicin. FROG and DD‐1 canine HSA cell lines were treated with varying concentrations of metformin or resveratrol with and without doxorubicin. Cell viability was measured by the MTT assay, and the expression of apoptotic proteins was assessed by western blotting. In contrast to human cell line studies, metformin had a minimal inhibitory effect on the growth of HSA cells. However, resveratrol markedly inhibited the growth of HSA cells, which is in agreement with human cell line studies. In combination, resveratrol and doxorubicin inhibited the growth of both HSA cell lines more than doxorubicin alone. Mechanistically, resveratrol induced apoptosis in HSA cells, and augmented doxorubicin‐induced apoptosis in these cells. Additionally, MTT cell viability of the cardiac‐derived H9c2 cells showed that resveratrol does not augment doxorubicin‐induced cell death in cardiac cells. These findings suggest that resveratrol may have anticancer effects against canine HSA and could be a potential adjunct therapy to the standard‐of‐care anticancer drugs used in canine HSA patients. Since resveratrol has been shown to have both anti‐cancer and cardioprotective effects in rodent models, canine cancer patients can be a perfect translational platform to move the findings of these rodent studies forward to the clinic.Support or Funding InformationResearch reported in this abstract was supported by a Pre‐K Career Development Award from the National Center for Advancing Translational Sciences of the National Institutes of Health Award Number UL1TR000114. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Effect of Prenatal Methamphetamine Exposure on Blood Pressure and Vascular Function in Adult Male and Female Offspring
Authors: Harrison Allison M. , Seeley Sarah L. , Rorabaugh Boyd R. , Chrissobolis Sophocles 
Published: November 2023

Hypertension and endothelial dysfunction are commonly associated with cardiovascular disease. Methamphetamine (meth) use is a widespread societal problem, with ~4.7% of the US population having tried meth at least once. Meth use during pregnancy can have deleterious developmental and behavioral consequences for offspring, including delayed development of gross motor skills and decreased memory. Although prenatal meth exposure reportedly causes increased cardiac ischemia in females, there exists very little additional data on the cardiovascular consequences of prenatal meth exposure in adult offspring. The aims of this study were to therefore test whether prenatal meth exposure increases blood pressure and impairs endothelial function in adult offspring, and whether this is influenced by gender. Pregnant female Sprague Dawley rats (8 wk old) were treated with either saline or methamphetamine (5 mg/kg/d sc, once daily) from gestational day 1 until pups were born. Male and female pups were weaned at 3 weeks of age and blood pressure (using tail cuff plethysmography) and vascular function (using myography) assessed after rats were 8 weeks old. Systolic blood pressure (SBP) was increased in male offspring prenatally exposed to meth (SBP = 151±2 mmHg, n=11) vs saline (141±3 mmHg, n=11, P<0.05 vs meth). SBP was similar in female offspring prenatally exposed to meth (SBP = 128±5 mmHg, n=7) and saline (SBP = 125±4 mmHg, n=8, P>0.05 vs meth). In males, endothelium‐dependent relaxation (assessed via relaxation responses to acetylcholine [ACh]) in second‐order mesenteric resistance arteries was impaired by prenatal meth exposure (pEC50: meth‐exposed = −7.5±0.06, n=5; saline‐exposed = −8.0±0.1, n=6; P<0.05). Prenatal meth exposure had no effect on responses to ACh in females (pEC50: meth‐exposed = −7.8±0.05, n=5; saline‐exposed = −8.1±0.08, n=5; P>0.05). Vasorelaxation responses to the endothelium‐independent agonist DEA NONOate, and phenylephrine‐induced vasoconstriction were not influenced by prenatal meth exposure in either gender. These data suggest that prenatal exposure to methamphetamine increases blood pressure and causes endothelial dysfunction in adult male offspring, whereas female offspring prenatally exposed to methamphetamine are protected against an increase in blood pressure and endothelial dysfunction. Prenatal methamphetamine exposure may therefore increase risk of cardiovascular disease in males.Support or Funding InformationRaabe College of Pharmacy, Ohio Northern University
Gestational Exposure to Engineered Nanomaterials Effects Microvascular Health of Young Progeny
Authors: Stapleton Phoebe A 
Published: November 2023

Engineered nanomaterials (ENM) are specifically manufactured at less than 100 nm in one dimension to take advantage of the unique physiochemical properties. While ENM are pervasive in domestic use, many are also being explored in biomedical applications as a platform for therapeutic or diagnostic (theranostic) applications. The toxicological implications of their use are tested primarily in healthy young adult male populations. However, these biomedical applications as contrast agents or directed drug delivery platforms may also be medically relevant during gestation. Unfortunately the outcomes during pregnancy and for developing young are unknown. In early studies, gestational ENM exposure led to impairments in uterine microvascular reactivity indicative of the development of a hostile gestational environment. The Barker Hypothesis theorizes that development is such an environment will influence cardiovascular health later in life.Therefore, studies were initiated to assess the microvascular health of fetal and postnatal young at differing developmental time points, exposed to nano‐sized titanium dioxide through maternal inhalation during gestation. Microvascular reactivity was assessed via a pressurized isolated microvessel preparation where the arterioles of the tail (fetal [gestational day 20], neonate [6 ± 0.48d]) or heart (weanling [25.7 ± 0.9d], juvenile [7.64 ± 0.48w], adult [10.36 ± 0.34w]) progeny were dissected, excised, and evaluated in response to chemical stimuli to calculate endothelium‐dependent [acetylcholine (10−9–10−4 M)], ‐independent [spermine‐NONOate (10−9–10−4 M)], and vascular smooth muscle [phenylephrine (10−9–10−4 M)] function. Significant temporal impairments in endothelium‐dependent reactivity were identified in fetal (−37.2% ± 7.3 vs. control, respectively), juvenile (−29.1% ± 11.3) and adult (−59.3% ± 17.1) exposed progeny. ­Following experiments performed with antioxidant treatment ([TEMPOL, 1 × 10−4 M] and catalase [50 units/mL]) in the isolated microvessel bath of the young adult cohort, led to an 86% ± 11.2 improvement in arteriolar reactivity and returned function to near control levels. These results indicate an increase in reactive oxidative stress (ROS) may be interfering in arteriolar function. Future studies may explore the origination of the ROS in the exposed progeny. Understanding the mechanisms associated with F1 effects may allow for pregnant women to safely reap the benefits of nanotechnology‐enabled products.Support or Funding InformationNIH‐K99‐ES024783; P30‐ES005022
Antenatal Betamethasone Fetal Programming of Cardiometabolic Disease Is Reversed by Removal of Visceral Adipose Tissue Early in Life
Authors: Shaltout Hossam A , Massmann Angela , Figueroa Jorge 
Published: November 2023

The use of antenatal betamethasone (BM) is a widely accepted therapy to improve survival in preterm infants. We have developed an experimental model of programming whereby pregnant ewes are administered BM (2 doses of 0.17 mg/kg) or vehicle at the 80th day of gestation and offspring deliver spontaneously at term. Our earlier studies show that BM‐exposed (BMX) offspring had a markedly impaired (~50% reduction) baroreflex sensitivity for control of heart rate (BRS) by 6 weeks of age followed by persistent suppression of the reflex and elevation in mean blood pressure (MAP) and development of glucose intolerance by 6 months of age compared to control group. In this study, we investigated the hypothesis that fetal programming of adipose tissue contributes to the elevation in MAP and reduction in BRS in BMX sheep. In a group of beta exposed sheep, Under general anesthesia, intra‐abdominal visceral fat (omental and perirenal) was surgically removed at 2–3 months of age, (BMX‐FR, n=7) and animals allowed to recover. At one year of age, heart rate and MAP were recorded continuously via femoral arterial catheters and BRS was tested using multiple doses of phenylephrine administered via femoral venous catheter to elevate MAP eliciting the reflex bradycardia. Data are presented as Mean ± SEM, and were analyzed by one way anova. While fat removal did not affect total body weight it lowered MAP by 20 mm Hg (from 95 ± 1 in BMX to 75 ± 4 in BMX‐FR, p < 0.05 to a level that is not different from the control group 78 ± 2 mm Hg). Also BRS improved by ~100 % as measured by ms/mm Hg (from 6 ± 1 in BMX to 13.8 ± 2.5 in BMX‐FR, p < 0.05 to a level that is not different from the control group 17 ± 3); there was no change in the heart rate between groups. At necropsy, we confirmed the effectiveness of the surgical fat ablation and found that BMX‐FR had a significant reduction in visceral fat expressed as % Body weight from 5.1± 0.5 to 1.0 ± 0.15. These data support the hypothesis that antenatal betamethasone exposure programs adipose tissue in a manner that leads to the impairment of cardiovascular system and that visceral fat removal early in life abrogate the cardiovascular dysregulation associated with betamethasone exposure. Further studies are being carried on to delineate the molecular changes in adipose tissues with fetal programming that leads to cardiometabolic disease and how it could be interrupted.Support or Funding InformationHD‐47584, HL 68728