bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolomic evaluation of Wistar rats' liver in experimental administration of 7,12‐Dimethylbenz(a)anthracene (DMBA)
Authors: Machado Vanessa Isabel , Cabral António , Tavares Ludgero , Farinha Rodrigo , Carvalho Rui , Cabrita António 
Published: June 2021

DMBA is a potent carcinogenic widely used in experimental oncology. It has a great mutagenic capacity after being metabolically activated in the liver and induces an accelerated tumor growth. In order to evaluate metabolic alterations associated with DMBA administration the metabolic profile of liver was analysed by high resolution magic angle spinning NMR spectroscopy (HRMAS), a powerful method to study how cellular physiology is affected by xenobiotics.Fifteen male wistar rats were divided into two experimental groups: control and DMBA administration Liver samples of male Wistar rats were collected and snap frozen in liquid nitrogen and later analyzed by high rotation magic angle NMR spectroscopy (HRMAS) in a in a 14.1 Tesla Varian spectrometer.HRMAS spectral analysis followed by principal component statistical analysis demonstrated that glycogen, glucose and insaturated lipids were significantly reduced in the DMBA group, while aspartate was increased.The results of this study suggest that DMBA reduces hepatic insaturated lipid contents and glycogen plus glucose. Increased glycolytic fluxes are characteristic of tumors and could explain the reductions in carbohydrate stores. Increased aspartate and reduced insaturated fatty acids could be the result of an alteration in cytosolic redox, which also characterizes tumorigenic activity.
ATP loss and purinergic receptor signaling contribute to early transcriptional responses activated by JNK in liver regeneration after partial hepatectomy in the rat
Authors: Juskeviciute Egle , Crumm Sara , Correnti Jason , Hoek Jan B. 
Published: June 2021

A complex program of gene expression changes drives the onset of liver regeneration after 70% partial hepatectomy (PHx) in the rat. However, the signals that initiate this response have not been well characterized. We reported recently (Crumm et al., 2008) that a rapid loss of adenine nucleotides (AdN) after PHx generates early stress signals that contribute to the onset of liver regeneration. Conditions that prevent the loss of AdN or that inhibit purinergic receptor activation suppress early signaling responses. This study focuses on the potential role of connexin (Cx) hemi‐channels in AdN loss and downstream signaling events. To specifically block different Cx isoforms, we used gap peptides. Pretreating animals with gap peptide specific for Cx26/32, but not for Cx43 or pannexin‐1 inhibited AdN loss. Pretreatment with the nitric oxide synthase inhibitor L‐NAME or the alpha‐adrenergic antagonist phentolamine also completely prevents loss of and upon PHx. However, pretreatment with the purinergic receptor antagonist suramin does not prevent AdN loss. All these treatments suppress Jun N‐terminal kinase (JNK) phosphorylation after PHx. In the analysis of downstream targets of JNK contributing to the early gene expression changes after PHx these inhibitors also decreased phospho‐cJun transcriptional activity at AP1 and CREB binding sites.Supported by NIH AA008714/AA018873 to JBH.
In vitro preliminary findings on the effect of biodegradable composite material in fibroblasts‐stimulated angiogenesis
Authors: Pirraco Ana , Rocha Ana Sofia , Vieira André , Vieira Joana , Guedes Rui , Soares Raquel 
Published: June 2021

The current ligaments treatment injuries has several drawbacks that prompted attempts to use synthetic materials in ligament replacements. Although fibroblasts do not comprise a large volume of ligament tissue, they have an important role in ligament tissue regeneration as they are able to model ECM and angiogenesis, therefore affecting the healing process.The aim is to appraise the biocompatibility of different biodegrable ligament composites (PGA‐PCL, PLA‐PCL, PDO, PGA) with the fibroblast‐mediated angiogenic process in vitro.HFF1 fibroblast cells were cultured on plates coated with each of the composite and cell viability was examined. Media from HFF1 cells incubated with composite material was removed and used to appraise the effect on angiogenesis (aortic ring assay and capillary‐like structures formation assay).No significant difference in cell viability was found for any composite tested. Aortic ring results suggested no disturbance of the angiogenic process. These findings were corroborated by the formation of cord‐like structures assay, as no significant difference in the number of structures was found in comparison to controls.Overall, our preliminary findings indicate that the composites do not promote nor prevent angiogenesis, being thus a good tool for the development of ligament composites. Supported by FCT (PTDC/EME‐PME/70155/2006)
Efficacy of Ozone Therapy Against the Formation of Peritoneal Adhesion in Rat
Authors: Uysal Bulent , Demirbag Suzi , Cayci Tuncer , Poyrazoglu Yavuz , Topal Turgut , Guven Ahmet , Korkmaz Ahmet 
Published: June 2021

INTRODUCTIONPeritoneal adhesion (PA) is a consequence of wound healing that sometimes causes severe postoperative complications. Despite all the preventive measures and agents, PA has still not been eliminated completely. The aim of this study was to evaluate the effect of the medical ozone therapy (MOT) on the development of postoperative peritoneal and uterine tube adhesions.MATERIAL AND METHODTwenty one female Wistar‐Albino type rats were used in this study. The rats were randomly divided into three groups of sham‐operated, PA and PA+MOT. After performing a lower midline abdominal incision, a standard uterine lesion was created by electrocauterization in the PA and PA+MOT groups. The PA+MOT group received 0.7 mg/kg per day ozone/oxygen mixture intraperitoneally for a total of 3 days. At the 7th day, peritoneal washing fluids of rats were obtained for TNF‐alpha evaluation. Then all of the rats were killed, and uterine adhesions were staged according to the Leach scoring system. Uterine tissue samples were harvested for biochemical and histopathological evaluation.RESULTSIn the PA+MOT group, the total adhesion score, oxidative stress markers including malondialdehyde, superoxide dismutase and glutathione peroxides were significantly lower than the PA group (P < 0.05). In addition, TNF‐alpha levels of peritoneal washing fluid were found decreased in the PA+MOT group than the PA group.CONCLUSIONMedical Ozone Therapy significantly reduces the prevalence of peritoneal and uterine tube adhesion formation in rats by possible decreasing oxidative stress.
Expression profiling of toll‐like receptor mRNAs in various tissues of Toda and Murrah buffaloes
Authors: Raj G Dhinakar , Venkatachalam Ramaswamy , Angamuthu Raja , Gopalan Tirumurugaan Krishnaswamy , Rathnassamy Vignesh Ambothi , Sakhthivel Dhanasekaran , Kathaperumal Kumanan 
Published: June 2021

Toda buffalo is a native genetically isolated semi‐wild breed, maintained by the aboriginal Toda tribes, present in the Nilgiri Hills of Tamil Nadu, India. The available population is only 1800. It is a brown medium sized animal with long horns set wide apart in a circular fashion. This breed resembles the swamp buffaloes, but based on karyological studies, it is classified under the river buffalo. The expression profile of toll‐like receptor (TLR) 1–10 mRNAs in spleen, mesenteric lymph node, lung, skin, jejunum and rectum were assessed using TaqMan real time PCR with GAPDH as endogenous control from Toda and Murrah buffaloes (n=4). The Ct values for different TLR mRNA in the tissues were compared among each breed with spleen expression levels as calibrator as well as between the two breeds and analyzed using the relative expression software tool (REST 2008‐V2.0.7). The Toda buffaloes showed higher expression levels of TLR 1 and 10 mRNA in the lymph node, TLR 1 in jejunum and TLR 1, 5, 6, 7 and 10 mRNAs in the skin. However, in Murrah buffaloes, lowered expression levels of TLR 10 mRNA in lung, TLR 4 in lymph node and TLR 10 in rectum and skin were observed. The Toda buffaloes showed significantly higher levels of TLR9 mRNA in the lymph node and TLR 1,5,6,9 and 10 mRNAs in the skin than Murrah. Thus, higher expression levels of a wide repertoire of TLR mRNA were detected in the skin of Toda buffaloes. The resistance of the Toda buffalo to protozoan parasites such as Theileria or Babesia, as is known anecdotally, could be related to the stronger innate immune responses mediated by TLRs in the skin.This work has been funded under the National Agricultural Innovative Project, Indian Council of Agricultural Research, New Delhi
The Effect of Quercetin in Cultured Human RPE Cells under Oxidative Stress and in Ccl2/Cx3cr1 Double Deficient Mice
Authors: Cao Xiaoguang , Tuo Jingsheng , Shen Defen , Liu Melissa , Chan Chi‐Chao 
Published: June 2021

PURPOSETo investigate mechanisms for the protective effects of quercetin in cultured human retinal pigment epithelium (RPE) cells and in Ccl2−/−/Cx3cr1−/− (DKO) mice, which spontaneously develop retinal lesions mimicking the characteristics of age‐related macular degeneration.METHODSCultured ARPE‐19 cells were exposed to 1mM H2O2 with or without 50μM quercetin for 2 hours. DKO and C57/B6 (WT) mice were treated with 25mg/kg/day quercetin via intraperitoneal injection for two months.RESULTSQuercetin protected RPE cells from H2O2‐induced injury, increased the Bcl‐2/Bax transcript ratio, decreased Bax, FADD, Caspase‐3, −9, TNF‐α, iNOS and COX‐2 mRNA levels, and lowered COX and NO levels. DKO mice had a lower antioxidative capacity, higher serum NO, COX and PGE2 levels, and higher ocular bax and caspase3 mRNA levels. Quercetin decreased serum oxidative and inflammatory molecules but not ocular pro‐apoptotic or pro‐inflammatory transcript levels; the retinal lesions in the DKO mice remained unaffected.CONCLUSIONSQuercetin protects human RPE cells from oxidative stress in vitro by directly inhibiting the apoptosis pathway and exerting anti‐inflammatory effects. Quercetin has no effect on retinal lesions in DKO mice due to insufficient suppression of ocular apoptosis or inflammation.The study was supported by the Intramural Research Program of the National Eye Institute, NIH.
Betaine‐homocysteine methyltransferase pharmacogenetics: human liver genotype‐phenotype association study
Authors: Feng Qiping , Kalari Krishna , Fridley Brooke L , Jenkins Gregory , Zhang Jianping , Johnson Monica Nye , Weinshilboum Richard M 
Published: June 2021

ObjectivesBetaine‐homocysteine methyltransferase (BHMT) catalyzes the remethylation of homocysteine. BHMT in the human liver is responsible for up to 25% of homocysteine metabolism. Knowledge of the relationship between genetic polymorphisms and BHMT phenotypes could enhance our understanding of the role of this reaction in drug metabolism.MethodsLevels of BHMT enzyme activity and immunoreactive protein were measured in 268 human liver biopsies. Twelve tag BHMT SNPs from our own gene resequencing and HapMap data were used to genotype DNA from these samples. Genotype‐phenotype association analysis was then performed. BHMT enzyme activity averaged 0.077 ± 0.028 units/μg, with a 12 fold variation.ResultsBHMT protein levels averaged 6.9 ± 3.3 units/μg, with a 50‐fold variation. Three SNPs (rs41272270, rs16876512, and rs6875201), located 28kb upstream, in the 5′‐UTR and in intron 1 of BHMT, respectively, were significantly correlated with both phenotypes (p<10−8 for enzyme activity and p<10−5 for protein level). rs41272270 was associated with altered protein binding during electrophoresis mobility shift assay.ConclusionsThree tag SNPs were significantly associated with human hepatic BHMT enzyme activity and immunoreactive protein. These observations may help us understand the regulation of BHMT in the human liver and its role in drug metabolism. Supported by NIH U01 GM61388, R01 GM28157, R01 CA132780.
Oral absorption and pharmacokinetics of Methadone HCl and intestinal multidrug resistance gene expression in horses
Authors: Linardi Renata Lehn , Barker Steven A , Short Charles , Kousoulas Konstantin G , Huang Li‐Ju , Stokes Ashley M 
Published: June 2021

This study aimed to determine the pharmacokinetics (PK) and oral bioavailability of methadone in horses to evaluate a possible influence of transport proteins on drug absorption through a relative quantitation of multidrug resistance gene/P‐glycoprotein (MDR1/P‐gp) gene expression in horse tissues.Six healthy horses received methadone by intravenous, oral, or nasogastric tube (NG) routes. Methadone serum concentration was measure and PK analysis performed. Total RNA was extracted from tissues from 10 horses and TaqMan RT‐PCR was used to quantify the gene of interest using Relative Standard Curve Method and fold‐differences in nucleic acid across tissues. Significance was determined with a P‐value < 0.05.Methadone was rapidly absorbed and eliminated with no side effects observed. Concentration was higher and occurred sooner after oral compared to the NG administration. Bioavailability was approximately 90% and 28% for oral and NG routes, respectively. Compared to the oral mucosa tissue, MDR1/P‐gp mRNA expression levels were significantly higher in the esophagus, stomach, small intestine, and liver.Methadone absorption from the small intestine appears limited possibly due to high expression levels of MDR1/P‐gp gene. High bioavailability after oral administration is supported by the low gene expression levels in tissues like oral mucosa and sublingual.Funding was provided by VCS and EHSP Grants, LSU.
Lysophospholipid signaling in human neural progenitors
Authors: Callihan Phillip , Gelineau‐van Waes Janee , Stice Steven L , Riley Ronald T , Hooks Shelley B 
Published: June 2021

Recent studies have shown that the fungal metabolite Fumonisin B1 (FB1) and the immunomodulatory drug FTY720 cause neural tube defects in mice. Both drugs inhibit ceramide synthase, resulting in an accumulation of sphingosine 1‐phosphate (S1P) and sphinganine 1‐phosphate (Sa1P) which are agonists for G‐protein coupled S1P receptors (S1PR). S1PRs are key regulators of neural development, suggesting that FB1 or FTY720 may cause developmental toxicity by altering signaling through S1PR. We have recently described S1P signaling pathways in human embryonic stem cell‐derived neuroepithelial cells (hES‐NEP). The goal of this project is to define the effects of FTY720 and FB1 on hES‐NEP cells, and establish this system as a platform in which to define the mechanism of lysophospholipid signaling in neural development. In the current study, we show that treatment of hES‐NEP with FB1 or FTY720 recapitulates the alterations in lipid profiles observed in‐vivo, resulting in marked increases in Sphinganine and Sa1P. Further, FTY720 is phosphorylated to the S1P receptor agonist FTY720‐P in hES‐NEP cells. Like S1P, both Sa1P and FTY720‐P stimulate activation of phospholipase C and inhibition of adenylyl cyclase, but with distinct rank order potencies and sensitivity to receptor and G‐protein inhibitors, suggesting these metabolites have unique signaling properties in human neural progenitors.Funding for this work is provided by the NIH and the University of Georgia.
Exercise is Associated with Increased Drug‐Taking in Rats that Self‐Administer Methamphetamine at Low Levels
Authors: Wright Mayo Jerry , Davis Sophia A , Dickinson Glen D , Taffe Michael A 
Published: June 2021

ObjectiveAbout 1% of young adults in the U.S. report using methamphetamine (METH) within the past year. Although METH is commonly used at dance parties, little is known about how physical activity influences use. Experiments were conducted to determine the relationship between voluntary exercise and METH self‐administration.MethodsRats were trained to self‐administer METH in chambers that contained both drug self‐administration apparatus and a running wheel. Rats were initially trained to self‐administer METH without wheel access. Once performance stabilized, rats were permitted access to the running wheel and the METH‐reinforced response lever concurrently.ResultsRats self‐administered METH at low levels prior to wheel access and the mean number of drug infusions taken during each session more than tripled after wheel‐access was permitted. Wheel running was stable within individual across sessions. The number of responses on the inactive lever was not significantly related to the dose of METH.ConclusionsIn these experiments, voluntary exercise was associated with increased METH self‐administration in rats that had previously demonstrated low levels of drug self‐administration. These results suggest that reinforcer competition is not the sole factor mediating effects of exercise on drug self‐administration. Research supported by National Institute of Drug Abuse grant R01 DA024705.
The differential expression of male sexual behavior in the Lewis, Fischer and Sprague‐Dawley rat strains
Authors: Hurwitz Zachary Edward , Riley Anthony Lee 
Published: June 2021

Lewis (LEW) and Fischer (F344) rats differ markedly in reactivity to drugs of abuse and mesolimbic dopamine activity, and interestingly sexual behavior is thought to be mediated in part by some common substrates. The following experiment examined sexual behavior of these strains (and outbred Sprague‐Dawley (SD) rats) to assess possible strain differences. Males were given repeated access to a receptive female of their respective strain during which sexual behavior was coded. Males were then exposed to a receptive LEW, F344 or SD female to assess if male behavior was a function of female receptivity. LEW males exhibited fewer mounts and intromissions (frequency and/or percent of subjects) than the F344 and SD strains (which never differed). Latency to first mount or intromission was similar for the F344 and SD strains but both were shorter than the LEW males (an index of sexual motivation). Ejaculation frequency and percent of subjects displaying ejaculation were different among strains (F344> SD>LEW). No LEW male ever ejaculated. For all males, there was no significant change in mounts, intromissions or ejaculations when given access to females of different strains, indicating the initial differences were not a result of strain differences in receptivity. These findings are consistent with a growing literature describing strain differences in motivated behaviors.Supported by a Mellon Foundation grant to ALR.
Effect of ovariectomy on the receptor mechanisms contributing to the discriminative stimulus effects of ethanol
Authors: Helms Christa Marie , Buckert Aubrey D , Grant Kathleen A 
Published: June 2021

The progesterone metabolites allopregnanolone and pregnanolone produce ethanol‐like discriminative stimulus effects in male Long‐Evans rats and in male and female cynomolgus monkeys. Fluctuations in endogenous ovarian‐derived neuroactive steroids alter ethanol and allopregnanolone substitution for ethanol in drug cynomolgus monkeys. This study determined whether absence of ovarian hormones alters the receptor mechanisms mediating the discriminative stimulus effects of ethanol. Sham (n = 13) and ovariectomized (OVX, n = 12) female Long‐Evans rats were trained to discriminate 1.0 g/kg ethanol (i.g., 15% w/v) from water with a 30‐min pre‐treatment interval. Ovariectomy increased pregnanolone and decreased 5‐HT receptor agonist substitution efficacy (Table 1). Compared to studies in males, neuroactive steroids showed lower substitution efficacy in female rats. Ovarian steroids in females affect specific receptor mechanisms mediating the discriminative stimulus effects of ethanol.
Percentage of rats showing complete substitution of test drugs for 1.0 g/kg ethanol and mean (± SEM) ED50 (mg/kg).

Sham
OVX

Ethanol
100
683 ± 37
100
620 ± 48

Allopregnanolone
54
6.8 ± 0.9
64
5.9 ± 1.1

Pregnanolone
38
6.3 ± 1.9
75
6.1 ± 1.7

Pentobarbital
85
5.1 ± 0.9
67
3.3 ± 0.7

Midazolam
75
1.2 ± 0.6
63
0.6 ± 0.3

Dizocilpine
62
0.036±0.006
88
0.036±0.008

TFMPP
62
0.6 ± 0.1
29
0.11±0.001

RU 24969
85
0.9 ± 0.1
57
0.4 ± 0.2

NIH AA017040
The Human Cytomegalovirus Encoded GPCR US28 Exhibits Constitutive Signaling in Productively Infected Glioblastoma Cells
Authors: Avery Diana L , Miller William E 
Published: June 2021

The Human cytomegalovirus (HCMV) is found in a high percentage of glioblastoma multiforme brain tumors. However, a causal relationship between the virus and glioblastoma has not been proven. Interestingly, HCMV encodes a gene termed US28 that is homologous to members of the GPCR superfamily. US28 functions as a constitutively active GPCR and is coupled to the Gαq/PLC‐β pathway. Recent studies indicate that US28 may activate pathways associated with cellular proliferation, suggesting that there may be a link between US28 and glioblastoma progression. In the present study, we aimed to utilize HCMV/US28 recombinant viruses to examine US28 expression and signaling activity in HCMV infected U373 glioblastoma cells. Our results indicate that US28 is expressed in HCMV‐infected glioblastoma cells with early kinetics and that it constitutively signals through the Gαq/PLC‐β pathway. These data support the hypothesis that constitutive signaling activity via the US28 GPCR could play a role in HCMV‐associated glioblastoma development. Future studies will need to define the downstream targets of US28 and determine if/how these targets regulate glioblastoma growth. These studies may facilitate the design of novel therapeutics targeted against this viral protein that could be ultimately used to slow or prevent HCMV‐associated disease.This project was supported by NIH Grant R01 AI058159 and by the ASPET SURF Program.
Modulation of Cannabinoid Receptor Interacting Protein 1a (CRIP1a) Expression Using Adeno‐Associated Virus and Effects on Striatal Neuropharmacology and Neuronal Cells in Culture
Authors: Blume Lawrence C. , Bass Caroline E. , Dalton George D. , Roberts David C.S. , Selley Dana E. , Howlett Allyn C. 
Published: June 2021

CB1 cannabinoid receptors and the D2 dopamine receptors are both GPCRs that associate with interacting proteins (CRIP1a and DRIP) that bind to the C‐terminus of the receptor. The DRIP neuronal calcium sensor‐1 is involved in Ca2+ mediated regulation of the D2 receptor, whereas the recently discovered cannabinoid receptor interacting protein (CRIP1a), has been shown to suppress the tonic inhibition of voltage‐gated Ca2+ channels induced by CB1 (Niehaus et al., 2007). Therefore, CRIP1a may provide a new avenue for modulation of the endocannabinoid system.AAV that contain transgenes to express shRNA designed to knock down CB1, D2R, or CRIP1a were created and injected via stereotaxically. Striata from AAV‐D2R and AAV‐CB1 shRNA‐treated rats displayed a significant reduction in D2 and CB1 mRNA levels, respectively. The AAV overexpressing CRIP1a virus increased CRIP1a mRNA levels without altering CB1 levels. However, significant reductions occurred in levels of preprodynorphin(Pdyn) and preproenkephalin‐1(Penk‐1).In vitro, N18TG2 stably transfected with CRIP1a‐pcDNA3.1 displayed increased levels of CRIP1a, Pdyn, and delta opioid receptor mRNA levels without effecting CB1 levels. Additionally, CRIP1a overexpressers had a significant reduction in CB1 mediated phosphorylation of ERK during Rimonabant‐sensitive basal conditions. However, CB1 agonist mediated effects on maximum ERK phosphorylation were unaltered in CRIP1a overexpressors.These studies provide both in vivo and in vitro insight into the mechanisms involved in CRIP1a modulation of CB1 receptors.