bluuebunny/crossref_metadata_2025_split huggingface dataset:
Semen Characteristics And Sperm Morphology Of Pistia stratiotes Linn. (Araceae) Protected Male Albino Rats (wistar strain) Exposed To Sodium Arsenite
Authors: Ola‐Davies Olufunke Eunice , Olawuwo Olasunkanmi Stephen , Ajani Olumide Samuel , Oyeyemi Olugbenga Matthew 
Published: November 2023

The sperm protective potential of Pistia stratiotes Linn (Aracea) was carried out in arsenic‐exposed rats using twenty‐four male albino rats (225 to 228g) aged between 14 and 16 weeks old. They were grouped into 4 (A–D), each group containing 6 rats. Group A animals were orally treated with 100mg/kg ethanol leaf extract of Pistia stratiotes Linn. daily for 14 days. B (sodium arsenite at 2.5 mg/kg body weight‐ positive control), C (Pistia stratiotes extract for 14 days and single dose of sodium arsenite on the 14th day, D (0.1 ml Propylene glycol‐ negative control/vehicle).Group B had a significantly lower (P<0.05) percentage sperm motility (26.7±6.67%) while group A had a significantly (P<0.05) higher mean value (63.3±3.33%) when compared across the groups. The sperm motility of rats in group D was significantly higher (P<0.05) than groups B and C. This implies that P. stratiotes extract had no adverse effect on sperm motility. The presence of P. stratiotes with sodium arsenite alleviated its harmful effect on sperm motility. The mean value obtained for sperm viability, semen volume and sperm count followed a similar pattern although, the difference was not significant (P>0.05) for semen volume and the sperm count of rats across the groups. Total sperm abnormality was 10.44 and 14.27% with the (SA) treated group having the highest value when compared with groups A treated with Pistia stratiotes extract and D treated with Propylene, although, the differences were not significant (P>0.05). The study concluded that ethanol leaf extract of Pistia stratiotes has no negative effect on sperm motility, viability and morphology and also protected spermatozoa against arsenic induced reproductive toxicity in wistar strain albino rats. Therefore, it may play an important role in the protection of populations with chronic sodium arsenite exposure.
The Marine Pharmacology and Pharmaceuticals Pipeline in 2015
Authors: Mayer Alejandro M , Nguyen Michelle , Newman David J , Glaser Keith B 
Published: November 2023

In October 2015 there were six FDA‐approved marine‐derived drugs: for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969), eribulin mesylate (Halaven®, FDA‐approved 2010), trabectedin (Yondelis®, FDA‐orphan drug approval 2005) being EU‐registered, and brentuximab vedotin (Adcetris®, FDA‐approved 2011); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004). The clinical marine pharmaceutical pipeline (Mayer et al. TIPS 31:255–265, 2010) consisted of 10 marine‐derived compounds in clinical development in 2010, including three monoclonal antibodies conjugated to monomethyl auristatin E, a synthetic analog of the marine compound dolastatin, in either Phase I, Phase II or Phase III clinical trials. The current clinical marine pharmaceutical pipeline consists of 9 marine‐derived compounds, and at least 16 other auristatin‐containing antibody drug conjugates in clinical development. Updated information on the clinical marine pharmaceutical pipeline is available at http://marinepharmacology.midwestern.edu/clinPipeline.htm. Finally, the global preclinical marine pharmacology pipeline was reviewed in Mayer et al. MARINE DRUGS 11:2510–2573, 2013). In late 2015, both the marine pharmacology preclinical and clinical pharmaceutical pipelines remained very active.Support or Funding InformationSupported by Midwestern University
EFFECT OF NIMESULIDE ON BLOOD SUGAR LEVEL AND HAEMATOLOGICAL INDICES OF ALBINO RATS
Authors: Agbasi Patrick Ugochukwu , Chukwu Leo Clinton , Unekwe Prince 
Published: November 2023

BackgroundNimesulide is relatively new drug that belong to the family of non steroidal anti inflammatory drugs (NSAIDs) and is a selective cyclo‐oxygenase 11 inhibitor that is gastro intestinal tract protective as against other non specific NSAIDs cyclo‐oxygenase inhibitors like indomethacin which usually cause stomach ulcer. NSAIDs are used as pain killers against post operative, menstrual, arthritic, cancer pains etc.PurposeThis study looked at the possible toxic effects of chronic administration of selecctive cyclo‐oxygenase 11 inhibitor like nimesulide on the haematological and fasting blood sugar level of Albino rats. The chronic use of selective cyclooxygenase 11 inhibitors in the treatment of musculo‐skeletal and joint pains of various origins necessitated this research.MethodsAlbino rats aged between four to five weeks were used. The animals were divided into three groups of five rats each. The first group received 5mg/kg/day, the second group also received 7.5mg/kg/day nimesulide respectively while the third received no drug and served as control. The drugs were orally administered for fifty six days during which the animals were given free access to feed and water. At the end of the study, the animals were anesthetized and blood samples were collected for biochemical and hematological parameters while spleen and pancreas were isolated for histological analysis.ResultsIt showed that the fasting blood sugar levels were not significantly increased p<0.05 when compared with control. The pancreas histopathology result produced no appreciable pathological changes in the two dose levels when compared with control. Also spleen histopathological result showed mild lymphoid reactive hyperplasia and moderate necrosis. The white blood cell (WBC) indices showed that total WBC count in 5mg/kg group were significantly increased p<0.05 when compared with control while the WBC level in 7.5mg/kg were significantly reduced p>0.05 when compared to both 5mg/kg rats and those of control. We recorded significant increase p<0.05 in lymphocyte level in both 5 and 7.5mg/kg group and significant reduction in mean monocyte level for both 5 and 7.5mg/kg rats when compared to control. The mean red blood cell (RBC) count values showed statistical significant increase P<0.05 in 7.5mg/kg Nimesulide group. Packed cell volume (PCV) values were not significantly increased p<0.05 in 5mg/kg group while the rats in 7.5mg/kg category were not significantly reduced when compared with control. Platelet values were statistically significantly increased p<0.05 in 5mg/kg group while those in 7.5mg/kg category were not significantly decreased p>0.05 when compared with control.Guiding Principles for the use and care of animals were strictly adhered to in this study.The Protocol for this research was approved by our institution's ethical committee.Support or Funding InformationSupport/Funding NON
Cholesterol 7α‐hydroxylase (CYP7A1) plays a critical role in preventing hepatic inflammation and fibrosis
Authors: Liu Hailiang , Ferrell Jessica , Boehme Shannon , Chiang John 
Published: November 2023

IntroductionCholesterol 7α‐hydroxylase (CYP7A1) is the rate‐limiting enzyme in the bile acid biosynthetic pathway in the liver and controls cholesterol and bile acid homeostasis. Accumulation of toxic bile acids in liver causes cholestasis and inflammatory liver injury. The mechanism of bile acid homeostasis in hepatic inflammation and fibrosis remains unclear.MethodsWe investigated the roles of CYP7A1 in hepatic inflammation and fibrosis using lipopolysaccharides (LPS)‐induced acute liver injury and methionine/choline deficient (MCD) diet‐induced liver injury models. Cyp7a1−/− mice in pure C57BL6 background, bile acid‐activated nuclear receptor farnesoid X receptor deficient (Fxr−/−) mice, and G protein‐coupled bile acid receptor deficient (Tgr5−/−) mice, and adenovirus‐mediated transduction of Cyp7a1 were used to study the roles of CYP7A1 and bile acid receptors in hepatic inflammation and fibrosis. Real‐time PCR, western blot, immunohistochemistry, luciferase report and chromatin immunoprecipitation (ChIP) assays were used in the study.ResultsAdenovirus‐mediated overexpression of CYP7A1 ameliorated LPS‐induced inflammatory cell infiltration and pro‐inflammatory cytokines production in WT and Tgr5−/−, but not Fxr−/− mice suggesting that FXR played a key role in protection against LPS‐induced hepatic inflammation. Luciferase report assay and real‐time PCR showed that FXR agonists, GW4064 or INT‐747 significantly inhibited NF‐κB activation and pro‐inflammatory cytokine mRNA expression in HepG2 cells and human primary hepatocytes. Mammalian two‐hybrid assay, co‐immunoprecipitation and ChIP assays showed that activation of FXR promotes FXR interaction with p65, which in turn blocked p65 binding to NF‐κB response elements on the TNF‐α and IL‐1β gene promoters. Serum AST and ALT levels were significantly increased in the liver of Cyp7a1−/− mice fed with MCD diet compared with WT mice. Pro‐inflammatory cytokine mRNA expression and immunohistochemistry of α‐SMA showed that Cyp7a1−/− mice fed with MCD diet developed more severe hepatic inflammation and fibrosis than WT mice. Cholesterol is aggravated in Cyp7a1−/− mice fed with MCD diet, and may increase reactive oxidizing species and cell death in the liver.ConclusionThese data suggest that increased bile acids ameliorated hepatic inflammation through activation of FXR but not TGR5 signaling. On the other hand, reduced bile acid pool increased susceptibility to MCD diet‐induced hepatic inflammation and fibrosis. Maintaining bile acid homeostasis is important in protection against liver injury and non‐alcoholic fatty liver disease.
Effect Of Ethanol On The Metabolism Of Antiretroviral Drugs Elvitegravir And Darunavir In The Presence And Absence Of Their Pharmacoenhancers In Microsomes
Authors: Midde Narasimha M , Rahman Mohammad A , Rathi Chetan , Li Junhao , Meibohm Bernd , Li Weihua , Kumar Santosh 
Published: November 2023

Alcohol consumption is highly prevalent in HIV positive individuals (50–70%) compared to the general population (14–24%). This drinking behavior is significantly associated with increased viral replication and decreased response to antiretroviral therapy (ART). Previously, we have shown that ethanol alters ART‐CYP3A4 interactions with protease inhibitors, which is likely to alter their metabolisms. In this study, we characterized Elvitegarvir (EVG), cobicistat (cobi)‐boosted EVG, Darunavir (DRV), and Ritonavir (RTV)‐boosted DRV metabolism in human microsomes followed by ethanol‐EVG/cobi and ethanol‐DRV/RTV interactions with CYP3A4. First, we developed and validated a simple, sensitive, and robust liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method for the quantification of EVG and DRV in the human liver microsomes. The lower limit of quantification for the EVG and DRV were at 0.003 μM (1.34ng/ml) and 0.023 μM (12.61ng/ml), respectively. Time kinetics data showed that 20 mM ethanol decreases the apparent half‐life of EVG metabolism by ~50%. Our substrate kinetic results revealed that ethanol slightly decreases the catalytic efficiency for EVG metabolism. Inhibition studies demonstrated that 20 mM ethanol causes a decrease in the IC50 of EVG. However, these parameters for EVG+cobi combination could not be determined because of strong inhibitory nature of cobi on CYP3A4. Furthermore, although ethanol did not show any appreciable effects on DRV kinetics with or without RTV, it robustly increased the IC50 of DRV in the presence of RTV. Finally, molecular docking studies predicted a shift of EVG or DRV binding parameters to the active site of CYP3A4 in the presence of ethanol. More specifically, the binding orientation of these substrates with CYP3A4 were altered in the presence of ethanol suggesting formation of new metabolite(s), which could be toxic. Taken together, these results suggest that ethanol interacts with microsomal CYP3A4 and alter ART metabolism, and their inhibition to CYP3A4. These findings have clinical implications with respect to adjustments of the regimens of these anti‐HIV drugs in alcohol users.Support or Funding InformationThis research was supported by grants from the National Institute of Health to Santosh Kumar (NIAAA/NIH AA‐022063) and the National Natural Science Foundation of China to Weihua Li (Grant 81373328)
Implications of In Vitro System Conditions for CYP2C9 and CYP3A4: Impact of pH on Clearance and Enzyme Inhibition
Authors: Rougee Luc R.A. , Mohutsky Michael A , Bedwell David W , Ruterbories Kenneth J , Hall Stephen D 
Published: November 2023

Common practice in drug discovery is to use buffer systems witha pH of 7.4 for drug metabolism and drug‐drug interaction studies despite evidence that the intracellular pH of hepatocytes is approximately 7.0. Systemp H will govern the ratio of ionized (charged) and non‐ionized (non‐charged)species for ionizable drugs. As a result of the pH gradient between the extracellular plasma and the intracellular cytosol of the hepatocyte, anunbound drug gradient, dependent on the physiochemical properties of the drug exists. An ionization correction factor [Berezhkovskiy (2011)]was proposed to account for this phenomenon when scaling in vitro findings at pH 7.4 to in vivo predictions of hepatic clearance. However, this correction ignores the potential changes in enzyme function at the lower pH environment.In the current study we evaluated, in human liver microsomes, the impact of buffer pH on the unbound fraction, Km, Vmax and intrinsic clearance (CLint=Vmax/Km) of probe substrates for two CYP enzymes, CYP2C9 (diclofenac and S‐warfarin) and CYP3A4 (midazolam, dextromethorphan, testosterone for CYP3A4) over a pH range of 6.0 to 9.0. Variable changes in the enzyme kinetic factors were observed over the pH range. These changes did not coincide with a particular species, ionizedor non‐ionized, for any of the substrates. When comparing the physiologically relevant pHs (7.0 and 7.4) for CYP2C9, a consistent trend was observed for both substrates, with lower Km, higher Vmax and higher CLint observed at 7.0 compared to 7.4. For the CYP3A4 enzyme, the inverse trend for observed, with higher Km, lower Vmax (Vmax was unchanged for midazolam) and lower CLint observed at 7.0 compared to7.4 for the dextromethorphan and midazolam substrates only. When testosterone was used as the probe substrate, both Km and Vmax were lower at pH 7.0 compared to pH 7.4, resulting in only a slight decrease in CLint at pH 7.0.Changes in the inhibition parameter, Ki, for the CYP2C9 reversible inhibitors amiodarone, desethylamiodarone, clozapine and nicardipine were consistent between the two probe substrates, with pH 7.0 Ki values consistently higher (less potent) than pH 7.4. Fold changes between pH 7.0 and pH 7.4 varied between the inhibitors and ranged from 1.25 to2.46 for the probe substrate diclofenac and 1.4 to 1.65 for S‐warfarin. These results suggest that changes as a result of pH vary from enzyme, compound and inhibitor combinations. In order to improve predictive performance of hepatic clearance, we recommend that in vitro parameters be determined at pH 7.0 to provide appropriate starting inputs for in vitro to in vivo extrapolations for ionizable drugs.
Identification of Novel Substrates and Substrate‐Dependent Interactions of Human Organic Cation Transporter 1
Authors: Boxberger Kelli H , Hagenbuch Bruno , Lampe Jed N 
Published: November 2023

The human organic cation transporter 1 (OCT1) is a polyspecific transporter involved in the uptake of positively‐charged and neutral small molecules in the liver. The clinical relevance of OCT1 has been established for several drugs, including the anti‐diabetic agent metformin. It has been demonstrated that OCT1 can contribute to drug efficacy, as well as to clinically relevant drug‐drug interactions (DDIs). Currently, the FDA recommends that the pharmaceutical industry screen new drug candidates for potential transporter‐associated DDIs using one or two probe substrates. Previous work from our laboratory suggests that inhibition of OCT1 may depend on the transported substrate. To study this further, we employed the competitive counterflow (CCF) efflux assay developed by Harper and Wright (Am J Physiol Renal Physiol. 2013;304(1):F56–67.) which tests a compound's ability to exchange with a known substrate. We have demonstrated that metformin and imatinib do not exchange with the probe substrate 1‐methyl‐4‐phenylpyridinium (MPP+) through OCT1. Because metformin and imatinib are known OCT1 substrates, these results indicate the possibility of different substrate binding domains and/or translocation pathways. Additionally, using the CCF assay, we have identified the negatively‐charged liver function test dye bromosulfophthalein (BSP) as a novel substrate of OCT1. These observed substrate‐dependent interactions may provide important insight into transporter structure and function. Furthermore, substrate‐dependent interactions can have important consequences when trying to identify DDIs for new drug candidates. Because regulatory guidelines only recommend one or two probe substrates to be tested, important DDIs may be missed. Therefore, these results may have important implications for revising guidelines for preclinical studies.Support or Funding InformationThis work was supported by the National Institutes of Health Institute of General Medical Sciences [Grant NIH R01‐GM077335] and [Grant P20‐GM103549‐07], National Center for Research Resources [Grant P20‐RR021940], National Institute of Environmental Health and Sciences [Grant T32‐ES007079], and Kansas IDeA Network of Biomedical Research Excellence [Grant QH846868‐K‐INBRE].
miR‐186 suppresses cell proliferation and anchorage‐independence in a metastatic prostate cancer cell line
Authors: Jones Dominique Z. , Schmidt M. Lee , Hobbing Katharine R. , Clark Geoffrey , Kidd LaCreis R. 
Published: November 2023

MicroRNA (miR) dysregulation alters the expression of cancer related genes and contributes to disease states in many cancers. For example, ectopic miR‐186 expression leads to enhanced cell proliferation and migration in pancreatic cancer. However, the role of miR‐186 in prostate cancer (PCa) remains unclear. Previously, we observed significant upregulation of miR‐186‐5p in PCa patient serum (stage III/IV) relative to non‐cancerous controls. Furthermore, miR‐186 was significantly up‐regulated in metastatic PCa (PC‐3) cells compared to normal prostate epithelial cells (RWPE1). We hypothesized miR‐186 inhibition will reduce aggressive PCa in vitro.Consequently, miR‐186 was transiently and stably inhibited in PC‐3 cells. Cell proliferation and colony formation were evaluated for 7 and 21 days via Trypan Blue exclusion and soft agar assays. Aberrant gene expression was evaluated in transfected cells to identify miR‐186 targets. Candidate miR‐186 targets were selected using published reports on ‘miR‐186 and cancer’, availability of robust antibodies, and statistical filtering (false discovery ≥0.05 and ±1.2 fold change).Mir‐186 inhibition in PC‐3 cells significantly repressed proliferation and colony formation by 34–64%. Following miR‐186 inhibition in PC‐3 cells, 2,343 identified mRNA targets were differentially expressed compared to scramble controls (p < 0.05). The target list was reduced to 11 up‐regulated candidates. Predicted miR‐186 targets are undergoing validation via qRT‐PCR, western blots, and luciferase reporter assays. In addition, other studies are in progress to evaluate the impact of miR‐186 on cellular migration and invasion. Such efforts may lead to the identification of novel biomarkers to improve detection and clinical management strategies for aggressive PCa.Support or Funding InformationResearch & Fellowship Support: National Cancer Institute Grant R25‐CA‐134283 to DWH; Our Highest Potential Endowment in Cancer Research to LRK
Novel ShetA2 Analog, SL‐01‐18, Triggers Apoptosis and Targets Androgen Receptors in Human Prostate Cancer Cells
Authors: Rajagopalan Vanishree , Nguyen Tran , Naizghi Girmay , Liu Shengquan 
Published: November 2023

SHetA2 (NSC 726189), a “Flexible Heteroarotinoid” compound that is presently in preclinical development for cancer prevention through the National Cancer Institute (NCI) Rapid Access to Intervention Development (RAID) program has shown to exert its anti‐cancer activity by regulating apoptosis, cell growth, differentiation and angiogenesis. However, SHetA2 may not be an ideal drug candidate due to the following two limitations: its high lipophilicity (Log P=7.9) and cumbersome six step synthesis. We have tried to address these limitations by designing over ten different analogs of SHetA2 by modifying the thiochromane ring of ShetA2 while keeping the nitrophenyl group and the thiourea linker intact. These analogs were tested for growth inhibition in three human prostate cancer cell‐lines. SL‐01‐18 was found to be the most potent growth inhibitor of prostate cancer cells. Further, SL‐01‐18 inhibited the growth of androgen dependent prostate cancer cell line, LNCaP, while minimally affecting the growth of androgen independent prostate cancer cell lines such as DU145 and PC‐3.The aim of this project was to further characterize the growth inhibitory effects of SL‐01‐18 on LNCaPs grown in media conditioned for androgen dependent growth. Within 24 hours, Hoechst 33342 live nuclear staining showed chromatin condensation and nuclear fragmentation, the hallmarks of apoptosis, in LNCaPs treated with SL‐01‐18. In vitro androgen receptor (AR) binding studies with SL‐01‐18 suggested that SL‐01‐18 might act as an antagonist to AR with an IC50 value of 2640nM. Since androgen receptor (AR) activation induces expression and secretion of prostate specific antigen (PSA), ELISA assays for both secreted and cellular PSA levels in LNCaPs revealed significantly lowered PSA levels after SL‐01‐18 treatment. Finally, a significant reduction in cellular AR levels in LNCaP cells was observed after treatment with SL‐01‐18 by western blot.These results confirm that SL‐01‐18 selectively inhibits growth of androgen dependent prostate cancer cells and eventually induces cell death via apoptosis. Our preliminary investigations into the mechanism of action of SL‐01‐18 in LNCaP cells reveal in vitro binding of SL‐01‐18 to AR and blocking it, potentially resulting in modulation of AR signaling by SL‐01‐18 as indicated by diminished AR protein as well as PSA levels upon treatment. Further mechanism directed studies will continue in future as will the design and synthesis of newer analogs SL‐01‐18.Support or Funding InformationThis research was supported by funds from TU‐COP MSMHS program.
The Repurposing of Ritanserin for the Inhibition of Diacylglycerol Kinase Alpha (DGKα), a Novel Therapeutic Target in Glioblastoma Multiforme (GBM)
Authors: Boroda Salome , Harris Thurl E. , Purow Benjamin W. 
Published: November 2023

Diacylglycerol Kinase α (DGKα) catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid (PA). Recent work has demonstrated that DGKα activity is at a central tumorigenic node and the attenuation of its activity by a small molecule inhibitor, R59022, has shown promising results in countering glioblastoma multiforme (GBM) growth both in vitro and in vivo (Dominguez, et al, 2013). Until recently, R59022 and R59949 have been the only described inhibitors of DGKα. We have identified a serotonin receptor antagonist, Ritanserin, which is almost structurally identical to R59022. Our data show that Ritanserin is a more potent inhibitor of DGKα activity than R59022 in vitro. We have additionally investigated the specificity of Ritanserin for DGKα over other mammalian DGKs, and the affinity of R59022 for serotonin receptors. Since Ritanserin has previously been used clinically, and shown to have a low incidence of adverse side‐effects, we believe that it could be a viable therapeutic agent for treatment of GBM. Additionally, our work sets the stage for the development for more potent DGKα inhibitors.Support or Funding InformationR01 1R01DK101946 to T.E.H, Wanger Fellowship Award, N.I.H Pharmacology graduate student training grant