bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chronic isolation stress alters antidepressant‐like behaviors in animals lacking RGS4
Authors: Talbot Jeffery N , Leon Bernice Poncede , Nielsen Cienna L , Benhatzel Christopher M , Alp Deniz , Kirsh Robert , Traynor John R 
Published: November 2023

Conventional treatments for depression increase serotonin (5‐HT) levels with selective serotonin reuptake inhibitors (SSRIs). Although superior to previous treatments, SSRIs cause unpleasant side effects, are ineffective in many patients, and take weeks to months for therapeutic benefit pointing to the need for improved understanding of mechanisms regulating mood and antidepressant drug action. Evidence suggests that antidepressant behaviors and more specifically the antidepressant actions of serotonin (5‐HT) are modulated by endogenous regulators of G protein signaling (RGS) proteins. In the current study, the effects of chronic isolation stress on mood‐related behaviors of mice lacking RGS4 were tested. Genetic deletion of RGS4 (RGS4(−/−)) had no effect on basal antidepressant‐like behaviors or generalized locomotor activity in mice. However, when subjected to chronic isolation stress, RGS4(−/−) mice developed antidepressant‐like phenotypes whereas unstressed RGS4(−/−) animals exhibited basal behaviors similar to stressed and unstressed wild‐type (RGS4(+/+)) littermates. Moreover, chronically‐stressed RGS4(−/−) animals exhibited increased phosphorylation of neuronal GSK3β, a putative biological marker of antidepressant‐like activity. Importantly, RGS4 deletion suppressed SSRI‐induced antidepressant activity but had no effect on basal or drug‐induced locomotor activity. Overall, these data suggest chronic stress differentially influences antidepressant‐like behaviors in an RGS4‐dependant manner and point to a role for RGS4 in the regulation of mood‐related behavior in vivo.Support or Funding InformationSupported by MH0907012.
Beta‐arrestin1 and testosterone levels in women with depression during premenstrual dysphoric disorder
Authors: Smith Keisha , Alam Farzana , Ansah Twum , Archibong Anthony , Nayyar Tultul 
Published: November 2023

OBJECTIVEWomen with premenstrual dysphoric disorder (PMDD), a severe form of premenstrual syndrome (PMS), have been suggested to have greater risk for future depression during pregnancy, post‐partum period and perimenopause. Currently no physical or laboratory test can diagnose PMDD. Reduction in beta‐arrestin1 (β‐AR1) protein and increased levels of testosterone are evident during depression. This study examined the levels of β‐AR‐1 protein in peripheral blood mononuclear leukocytes (PBMC) and plasma testosterone in women with PMDD as diagnosed by objective assessment from questionnaires, which may aid in the diagnosis and treatment for PMDD.DESIGN METHODSStudy participants (n=25) were non‐pregnant women between 18–42 years of age with the symptoms of PMS/PMDD, but not taking any antidepressants/therapy and at the luteal phase of menstruation. The protein levels of beta‐arrestin1 in the PBMC were determined by ELISA and the testosterone levels in plasma were measured by radioimmuno assay. The participants were evaluated with Neuropsychiatric Interview (DSMIV‐TR). The severity of depression were determined by the Hamilton Depression Rating Scale scores (HRSD).RESULTSThe magnitude of the different parameters for Axis 1 mental disorders and the plasma testosterone levels were significantly higher and beta arrestin1 protein levels in mononuclear leukocytes were significantly lower in women with depression (PMDD) as compared to PMS women.CONCLUSIONThese findings support that reduction in beta‐arrestin1 protein and associated increased levels of testosterone are implicated in the pathophysiology of depression in women with PMDD.Support or Funding InformationClinical Research Education and Career Development in Minority R25RR17577 (NCRR/NIMHD) and U54 MD007593 (NIMHD) and U54 RR026140 (NCRR) at NIH
Role of Serotonin 2A and 2C Receptors in Ketamine‐induced Hypothermia
Authors: Murphy Tyler Joseph , Murnane Kevin Sean 
Published: November 2023

Anesthesia‐Induced Hypothermia (AIH) has been reported to cause many postoperative adverse effects. These effects include increased mortality, immunosuppression, increased risk of cardiac events, and a higher prevalence of surgical wound infections. Currently, there are few effective ways of preventing AIH besides pre‐warming fluids and gases for the patients during surgery, or relying on forced‐air heating systems. Although it is established that serotonin has a role in thermoregulation, the receptors responsible for such effects remain unclear. Here, we investigated the role of serotonin receptors in AIH using the 5‐HT2A receptor antagonist M100907 and the 5‐HT2C receptor agonist WAY‐163909. Core body temperature was determined through the use of a rectal probe and hypothermia was induced by ketamine in male, young adult Swiss‐Webster mice. We found that dose‐dependently and reliably ketamine induced hypothermia. Moreover, WAY‐163909 attenuated ketamine‐induced hypothermia. In contrast, ketamine‐induced hypothermia was exacerbated by M100907. These findings suggest a new clinical use for 5‐HT2C receptor agonists, specifically to maintain body temperature during surgery. We will build upon these studies by conducting mechanistic pharmacological and neurobiological experiments, including immunohistochemical assays to determine the effects of these drugs on activity in the preoptic area of the anterior hypothalamus, the thermoregulatory center of the brain.Support or Funding InformationThis work was supported by institution funds supplied by the Mercer University College of Pharmacy, an honors thesis award supplied by Oglethorpe University, and a Summer Undergraduate Research Fellowship supplied by the American Society for Pharmacology and Experimental Therapeutics.
Synthesis of chitosan nanoparticles of flavonoids extracted from black bean (Phaseolus vulgaris L.) seed coats to enhance the intestinal absorption and increase plasma residence time
Authors: Sanchez‐Ramirez Carlos , Chavez‐Santoscoy Rocio Alejandra , Cornejo‐Bravo Jose Manuel 
Published: November 2023

Beans exert hypolipidemic activity, and it has been attributed to flavonoids and saponins from seed coats, by attenuating lipogenesis and stimulating fatty acid oxidation and cholesterol biliary excretion through the differentially regulation of LXR probably mediated by AMPK phosphorylation. However, their therapeutic potential is limited by their low oral bioavailability, attributed to poor stability and intestinal absorption. Chitosan (CS) is a natural biodegradable, biocompatible and non‐toxic biopolymer extracted from the shells of crustaceans. Chitosan and its derivatives have been examined extensively for medical and pharmaceutical applications such as targeted drug delivery, drug transport, and so on. The chitosan tripolyphosphate (CS/TPP) nanoparticles have been used as an alternative to encapsulate bioactive flavonoids from black bean seed coats. The particle size measurements of the nanoparticles were measured by the Zetasizer Nano and the cytotoxicity of CS/TPP nanoparticles was evaluated by MTT assay using fibroblasts from Mus musculus (NIH/3T3 cell line). Nanoparticles size was approximately 150 nm, the encapsulation efficiency of CS/TPP nanoparticles was found to be 59.9%, and they were not cytotoxic at a concentration of 500mg/mL in fibroblast cells. CS/TPP naanoparticles are an interesting alternative to develop a delivery systems for flavonoids and natural bioactive compounds.Support or Funding InformationThis research was supported by the Research Chair Funds for Full Time Professors (2014) of Autonomous University of Baja California (UABC), program Number 3888. The authors would like to acknowledge Consejo Nacional de Ciencia y Tecnología (CONACYT) in Mexico.
Induction of Nrf2 Activity by Ethyl Acetate Extract of Lagenaria Breviflora
Authors: Oridupa Olayinka Ayotunde , Ritchie Kenneth J 
Published: November 2023

The field of cancer chemoprevention is based upon the ability of chemical compounds from any source (synthetic and natural) to prevent cancer. Phytochemicals with the ability to increase the activity of the transcription factor Nrf2 (NF‐E2‐related factor 2) are central to this field. Nrf2 has been reported to modify the expression of up to 200 genes, many of which are involved in cellular defence against toxic insult including heme‐oxygenase 1 (HO‐1), NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S‐transferases (GSTs) [1]. Interestingly many plant species which are known possess direct antioxidant activity through mechanisms such as free radical scavenging have not been investigated for their ability to modify the expression of such crucial cellular defence genes. This study was consequently designed to investigate the Nrf2 induction capability of the medicinal plant Lagenaria breviflora Roberty (LB) which is documented to possess both in vivo and in vitro antioxidant and anti‐inflammatory activities [2].Nrf2 induction was investigated using the AREc32 cell line, (Nrf2 luciferase assay reporter cell line) and by activation of the Nrf2 regulated gene NQO1 in HepG2 cells by Western blot. Exposure of AREc32 cells to graded concentrations of LB ethyl acetate extract (4mg/ml, 6mg/ml, 8mg/ml and 12mg/ml) significantly increased luciferase activity by 3 – 7.5 fold relative to control. Higher concentrations (8mg/ml and 12mg/ml) were however toxic to the cells (by MTT assay). Kinetic studies revealed significant induction of luciferase activity in AREc32 cells treated with 6mg/ml of the extract for 24 hours (4 fold) and 72 hours (3 fold). LB extract (6mg/ml) also significantly increased the expression of NQO1 protein in HepG2 cells, relative to control, following 24 hour exposure. These results indicate that the ethyl acetate extract of LB contains Nrf2 modulating activity and as such indicates that LB is worthy of further investigation into its potential to act as a cancer chemopreventative agent.Support or Funding InformationThis research was supported by TETFund Staff Development Grant (Nigeria) Awarded to Dr. Olayinka A. Oridupa
Characterization of Nitrogen Mustard‐Induced Lung Injury Using Molecular Imaging Techniques
Authors: Venosa Alessandro , Schumacher Justin , Adler Derek , Laskin Jeffrey , Yurkow Edward , Gow Andrew , Laskin Debra 
Published: November 2023

Nitrogen mustard (NM) is an alkylating agent known to cause acute lung injury, which progresses to fibrosis. High definition magnetic resonance imaging (MRI) and computed tomography (CT) were used to follow the progression of NM‐induced injury. Male rats (8–10 wk) were treated i.t. with 0.125 mg/kg NM or control. Gradient (GRE) and fast spin echo (FSE) MRI, and CT scans were performed 1–28 d later. Image processing was used to analyze and quantify MRI scans for areas of lung injury. NM exposure was associated with an increase in lung injury volume which was evident within 1–3 d; this coincided with acute injury in histologic sections. Whereas histology and bronchoalveolar lavage fluid analysis indicated a time related decrease in extravascular lung fluids, MRI imaging showed low‐density shadows, indicative of fluid buildup, up to 28 d. CT scans were used to generate a high definition 3D reconstruction of the lung. Based on pixel intensity, lung images were segmented and the volume of air, dynamic and consolidated lung tissue identified. Within 3 d of NM dynamic lung volume was significantly decreased, when compared to controls, while the volume of consolidated lung tissue increased with time. These data show that MRI and CT imaging effectively track the progression of NM‐induced lung injury and have the potential to be used to identify a signature of acute injury and fibrosis.Support or Funding InformationNIH AR055073, HL096426, ES004738, ES005022
Altered Ion Transport in Normal Human Bronchial Epithelial (NHBE) Cells Following Exposure to Metal Welding Particles
Authors: Fedan Jeffrey S , Thompson Janet A , Meighan Terence G , Erdely Patti C , Antonini James M 
Published: November 2023

Inhalation of welding fumes is associated with adverse effects in the pulmonary system, including susceptibility to infection. Several types of welding processes occur, giving rise to fumes of different compositions. In this study we investigated the effects of particles derived from fumes generated from manual metal arc‐stainless steel (MMA‐SS) and gas metalarc‐mild steel (GMA‐MS) welding on ion transport in NHBE in culture. The toxicities and the composition of the particles generated during MMA‐SS and GMA‐MS processes are different. MMA‐SS particles are more soluble and contain Cr, Ni, Fe and Mn, whereas GMA‐MS particles are less soluble and contain Fe and Mn. Ion transport is critical to the maintenance of the airway surface liquid. We hypothesized that epithelial cells are targets of inhaled welding fumetoxicity. NHBE (Lonza) were cultured in submersion culture for 3 days followed by 21 d in air‐interface on permeable inserts; cells with transepithelial resistance (Rt) >1000 Ω·cm2were used. Generated MMA‐SS or GMA‐MS particles collected on filters were suspended in medium and applied (50 μl) to the apical chamber. The doses of both types of particles ranged from 0.0167 to166.7 μg/cm2. After 18 h of incubation the inserts were placed in Ussing chambers for the measurement of transepithelial potential difference (Vt), short circuit current (Isc) and Rt. The effects of fume exposure on Na+ and Cl− channels, the Na+,K+,2Cl−‐cotransporter and the Na+,K+‐pump, were evaluated by applying the following inhibitors, respectively: amiloride(A; apical; 3×10−5 M), NPPB (N; apical; 10−4 M), bumetanide (B; 10−4 M) and ouabain (O; basolateral; 10−4M). Changes in lactate dehydrogenase(LDH) levels in apical medium also were assessed after exposure. Following exposure, 0.0167–16.7 μg/cm2MMA increased basal Vt. Only16.7 μg/cm2 GMA‐MS increased basal Vt. MMA‐SS exposure potentiated Isc responses (decreases in Isc) to A and B, while not affecting those to N and O. Likewise, exposure to GMA‐MS potentiated Isc responses to A and B, but to a lesser degree than observed after MMA‐SS exposure. Variable effects on Rt were observed in response to A, B and O. Generally, MMA‐SS was more potent in altering responses to A and B than GMA‐MS. MMA‐SS evoked greater increases in LDH release than GMA‐MS, but bioelectric changes occurred in the absence of increased LDH release. Our findings indicate that both apical Na+ transport and basolateral Na+,K+,2Cl−‐cotransportin airway epithelium is affected by fume particle exposure. The greater effect of MMA‐MS compared to GMA‐MS might reflect the actions of soluble metals. Enhanced Na+ absorption and decreased airway surface liquid could compromise lung defenses against infection.Support or Funding InformationSupported by NIOSH
Mechanistic Role of Cytochrome P450 (CYP)1A2 in the Regulation of Hepatic and Pulmonary CYP1A1 Expression in Mice Treated with 3‐Methycholanthrene
Authors: Moorthy Bhagavatula , Maturu Paramahamsa , Wang Lihua , Chu Chun , Jiang Weiwu 
Published: November 2023

3‐Methylcholanthrene (MC) is one of most potent polycyclic aromatic hydrocarbons (PAHs) present in cigarette smoke, diesel exhausts, and charcoal broiled meats, etc. Cytochrome P450 (CYP) 1A enzymes play key roles in the activation of PAHs to carcinogenic metabolites. In this study, we tested the hypothesis that CYP1A2 plays a mechanistic role in the regulation of hepatic and pulmonary CYP1A1 by MC. Wild type (WT) (C57B6) and Cyp1a2_null mice were divided into two groups. Group I was treated with vehicle corn oil (CO) (8 ml/kg) and group II was treated with four doses of MC (100 μmol/kg), i.p. once daily for 4 days. Four animals from each group were sacrificed at 6, 12, 24, and 48 h, 8 and 15 days after MC withdrawal. The mRNA levels, protein content and enzyme activities of CYP1A1 and 1A2 were determined at different time points. In addition, the binding of MC‐AHR‐AHR nuclear translocator (ARNT) to the AHREs on the CYP1A1 promoter region was determined by chromatin immunoprecipitation (ChIP) assay in liver and lung tissues. In WT mice, ChIP experiments indicated that transcriptional activation of CYP1A1 was most pronounced at 6 h, followed by 12 h, but declined at later time points. In Cyp1a2_null mice, on the other hand, transcriptional activation of CYP1A1 persisted till 48 h. In lungs, AHR complex binding peak was observed at 12 h and declined later, but Cyp1a2 null mice showed more persistent binding till 48 h with a peak at 24 h. On the other hand, the expression of CYP1A1/1A2 at the mRNA, protein, and enzyme levels persisted for up to 48 h both in lung and liver tissues. These results suggest a mechanistic role for CYP1A2 in the molecular regulation of CYP1A1, and that transcriptional activation of CYP1A1 at 6–12 h is sufficient to result in sustained induction of CYP1A mRNA and protein expression for up to 48 h. These studies have important implications for carcinogenesis mediated by PAHs.Support or Funding InformationNIH Grant R01 ES009132
Effect of CYP17A1 17,20 Lyase Inhibitors on Regulation of Adrenal Androgen Biosynthesis
Authors: Pandey Amit V , Udhane Sameer S 
Published: November 2023

BackgroundThe P450c17 (CYP17A1) plays a vital role in regulating adrenal androgen production. CYP17A1 localized in the endoplasmic reticulum can catalyze both 17α‐hydroxylase and 17,20 lyase reactions. Understanding the mechanisms regulating 17,20 lyase activity is important for the understanding of hyperandrogenic disorders such as premature, exaggerated adrenarche and the polycystic ovary syndrome, and also for the design of selective 17,20 lyase inhibitors for use in hyperandrogenic states and in sex‐steroid dependent cancers. The orteronel and galeterone are known to inhibit 17,20 lyase activity; however, the precise mechanism of the CYP17A1 inhibition remains unknown. These inhibitors have been developed to treat the castration resistant prostate cancer (CRPC) but little is known about their effect on adrenal androgen biosynthesis. We have studied the specific 17,20 layse inhibitors to understand their effects on CYP17A1 and adrenal androgen biosynthesis.MethodsWe used NCI‐H295R adenocarcinoma cells to study the effect of orteronel and galeterone. We treated the H295R cells with 0–2μM orteronel and galeterone for 24 hours. Steroid production was labeled with [3H] pregnenolone for 90 min. Steroids were extracted and resolved by thin layer chromatography. For specific analysis of the CYP17A1, CYP19A1 and CYP21A2 activities, cells were treated with 1μM trilostane (a specific blocker of HSD3B) for 90 min before adding radiolabelled steroids. To study the effect of these inhibitors on steroidogenic gene expression we performed the relative quantification PCR (qRT‐PCR). Steroid metabolome of cells was determined in parallel experiments using GC‐MC/LC‐MS.ResultsThe drugs orteronel and galeterone were able to inhibit CYP17A1 activity in H295R cells. Both drugs have more potency towards the 17,20 lyase activity but we also observed that they partially affected 17 alpha‐hydroxylase activity. From our results, we observed the orteronel seems to be more potent and selective towards 17,20 lyase activity than galeterone. However, we also found dihydroepiandrosterone (DHEA), cortisol and androstenedione were drastically decreased by both compounds at 1 and 2μm concentration. Further, we also studied effect of these inhibitors on genes which are involved in androgen biosynthesis. We found slight increase in the HSD3B2 gene expression under galeterone treatment but no change was observed in CYP17A1 and AKR1C3 expression.ConclusionsBased on our results we conclude that orteronel is a more potent inhibitor of 17,20 lyase activity and also has partial effect on 17α‐hydroxylase activities.Support or Funding InformationThis work was supported by grants to Amit V Pandey from the Swiss National Science Foundation and Bern University Research Foundation.
Inhibition of Human CYP3A‐Catalyzed Lithocholic Acid 3‐Oxidation by Specific Vitamin E Isomers
Authors: Lau Aik Jiang , Chai Swee Fen , Teo Josephine Si Min 
Published: November 2023

Lithocholic acid is a hepatotoxic and carcinogenic bile acid. Previous studies suggested that CYP3A4 but not CYP3A5 catalyzed the oxidation of lithocholic acid to 3‐ketocholanoic acid and that CYP3A metabolized vitamin E isomers (tocotrienols and tocopherols). Given that a substrate of an enzyme may also be an inhibitor of that enzyme, we determined whether α‐tocotrienol, γ‐tocotrienol, δ‐tocotrienol, a tocotrienol‐rich mixture (a mixture consisting of 25.7% α‐tocotrienol, 2.6% β‐tocotrienol, 28.6% γ‐tocotrienol, 8.4% δ‐tocotrienol, 25.6% α‐tocopherol, and 4.3% α‐tocomonoenol), and α‐tocopherol inhibit the oxidation of lithocholic acid to 3‐ketocholanoic acid. Enzymatic formation of 3‐ketocholanoic acid via lithocholic acid 3‐oxidation was determined in pooled human liver microsomes and recombinant CYP3A4 and CYP3A5 enzymes. Enzyme inhibition assay was conducted in an incubation mixture containing potassium phosphate buffer, lithocholic acid, NADPH, enzyme (human liver microsomes, recombinant CYP3A4, or recombinant CYP3A5), and a putative inhibitor (a tocotrienol, a tocotrienol‐rich mixture, or α‐tocopherol). The amount of 3‐ketocholanoic acid was quantified by ultra‐high performance liquid chromatography‐tandem mass spectrometry. Lithocholic acid was metabolized to 3‐ketocholanoic acid by human recombinant CYP3A4 and CYP3A5 enzymes and human liver microsomes. Human liver microsomal‐catalyzed lithocholic acid 3‐oxidation exhibited Michaelis‐Menten kinetics at substrate concentrations less than 100 μM. Substrate inhibition kinetics was evident at lithocholic acid concentrations greater than 100 μM. The Km, Vmax, and Vmax/Km values for human liver microsomal LCA 3‐oxidation were 26 μM, 304 pmol/min/mg, and 12 μl/min/mg, respectively. Enzyme kinetics experiments indicated that human recombinant CYP3A4 and CYP3A5 catalyzed lithocholic acid 3‐oxidation to a similar extent. α‐Tocotrienol, γ‐tocotrienol, δ‐tocotrienol, and a tocotrienol‐rich mixture, but not α‐tocopherol, inhibited lithocholic acid 3‐oxidation by human liver microsomes. Concentration‐response experiments indicated that a tocotrienol‐rich mixture and δ‐tocotrienol inhibited human liver microsomal lithocholic acid 3‐oxidation with IC50 of 6.6 ± 2.1 μg/ml and 19.0 ± 1.0 μM, respectively. Overall, lithocholic acid 3‐oxidation is catalyzed by both CYP3A4 and CYP3A5 enzymes. It is differentially inhibited by tocotrienols, but not α‐tocopherol, suggesting vitamin E isomer‐specific inhibition of human liver microsomal lithocholic acid 3‐oxidation.Support or Funding InformationSupported by National University of Singapore Research Fund.