Miracle-dz/UniFAR-FTUs huggingface dataset:
Inhibition of TGF- b signaling by an ALK5 inhibitor protects rats from dimethylnitrosamine-induced liver fibrosis

1 Chronic liver disease is characterized by an exacerbated accumulation of matrix, causing progressive fibrosis, which may lead to cirrhosis. Transforming growth factor beta (TGF- b ), a well-known profibrotic cytokine, transduces its signal through the ALK5 ser/thr kinase receptor, and increases transcription of different genes including PAI-1 and collagens. The identification of GW6604 (2-phenyl-4-(3-pyridin-2-yl-1 H -pyrazol-4-yl)pyridine), an ALK5 inhibitor, allowed us to evaluate the therapeutic potential of inhibiting TGF- b pathway in different models of liver disease. 2 A cellular assay was used to identify GW6604 as a TGF- b signaling pathway inhibitor. This ALK5 inhibitor was then tested in a model of liver hepatectomy in TGF- b -overexpressing transgenic mice, in an acute model of liver disease and in a chronic model of dimethylnitrosamine (DMN)-induced liver fibrosis. 3 In vitro , GW6604 inhibited autophosphorylation of ALK5 with an IC 50 of 140 n M and in a cellular assay inhibited TGF- b -induced transcription of PAI-1 (IC 50 : 500n M ). In vivo , GW6604 (40mg kg (cid:1) 1 p.o. ) increased liver regeneration in TGF- b -overexpressing mice, which had undergone partial hepatectomy. In an acute model of liver disease, GW6604 reduced by 80% the expression of collagen IA1. In a chronic model of DMN-induced fibrosis where DMN was administered for 6 weeks and GW6604 dosed for the last 3 weeks (80mg kg (cid:1) 1 p.o. , b.i. d. ), mortality was prevented and DMN-induced elevations of mRNA encoding for collagen IA1, IA2, III, TIMP-1 and TGF- b were reduced by 50–75%. Inhibition of matrix genes overexpression was accompanied by reduced matrix deposition and reduction in liver function deterioration, as assessed by bilirubin and liver enzyme levels. 4 Our results suggest that inhibition of ALK5 could be an attractive new approach to treatment of liver fibrotic diseases by both preventing matrix deposition and promoting hepatocyte regeneration. British
PTB’S NEW 200 KN DEADWEIGHT FORCE STANDARD MACHINE

This paper describes PTB’s new 200 kN deadweight force standard machine with a large force range from 50 N up to 200 kN with lowest relative uncertainties of 0.001% in the range from 5 kN up to 200 kN which was possible by using deadweights of stainless steel in the quality of mass standards of class F1. The deadweights are adjusted according the gravity value and the masses are then determined with a relative expanded uncertainty of 3E-6. This maschine allows the investigation and calibrate force transducers in tension and compression without changing the installation position so that it is possible to determine the hysteresis effect (remanance) if the force changes from tension to compression.
Interaction of the Za domain of human ADAR1 with a negatively supercoiled plasmid visualized by atomic force microscopy

Interest to the left-handed DNA conformation has been recently boosted by the findings that a number of proteins contain the Za domain, which has been showntospecifically recognize Z-DNA.The biological function of Za is presently unknown, but it has been suggested that it may specifically direct protein regions of Z-DNA induced by negative supercoiling in actively transcribing genes. Many studies, including a crystal structure in complex with Z-DNA, have focused on the human ADAR1 Za domain in isolation. We have hypothesized that the recognition of a Z-DNA sequence by the ZaADAR1 domain is context specific, occurring under energetic conditions, which favor Z-DNA formation. To test this hypothesis, we have applied atomic force microscopy to image ZaADAR1 complexed with supercoiled plasmid DNAs. We have demonstrated that the ZaADAR1 binds specifically to Z-DNA and preferentially to d(CG)n inserts, which require less energy for Z-DNA induction compared to other sequences. A notable finding is that site-specific Za binding to d(GC)13 or d(GC)2C(GC)10 inserts is observed when DNA supercoiling is insufficient to induce Z-DNA formation. These results indicate that ZaADAR1 binding facilities the B-to-Z transition and provides additional support to themodel thatZ-DNA binding proteinsmayregulate biological processes through structure-specific
Regulation of STREX BK channels by the β 4 Accessory Subunit

Large conductance (BK-type) calcium-activated potassium channels utilize alternative splicing and association with accessory β subunits to tailor BK channel properties to diverse cell types. Two important modulators of BK channel gating are the neuronal-specific β 4 subunit and alternative splicing at the STREX exon. Individually, these modulators affect the gating properties of the BK channel as well as its response to phosphorylation. In this study, the combined functional consequences of STREX and the β 4 subunit on BK channel biophysical properties were investigated in transfected HEK293 cells. Surprisingly, we found that the combined effects of STREX and β 4 are non-additive and even opposite for some properties. At high calcium, β 4 or the STREX exon individually share properties that promote BK channel opening via slowing of deactivation. However, the combined effects are a speeding of deactivation and a decreased open probability. β 4 also inhibits BK channel opening by a slowing of activation. This effect occurs across calcium concentrations in the absence of STREX, but predominates only at low calcium for STREX containing channels. BK channel response to phosphorylation status are also altered by the combination of the β 4 subunit and the STREX exon. β 4/STREX channels show a slowing of activation kinetics following dephosphorylation whereas β 4 channels lacking STREX do not. In contrast, β 4 confers a speeding of activation in response to cAMP-dependent phosphorylation in channels lacking STREX, but not in channels containing the STREX exon. These results indicate that the combination of the β 4 subunit and STREX exon confer non-additive and unique properties to BK channels. Analysis of expression in brain slices suggests that STREX and β 4 mRNA overlap expression in the dentate gyrus of the hippocampus and the cerebellar purkinje cells, suggesting that these unique properties of BK channels may underlie BK channel gating in these cells.
Prize-based contingency management is efficacious in cocaine abusers with and without recent gambling participation

Prize-based contingency management (CM) is efficacious in treating cocaine abuse, and the chance-based procedures of prize CM may be appealing to those who gamble. Using data from three randomized trials, we evaluated whether cocaine abusers who had wagered in the month before treatment ( n = 62) responded more favorably to prize CM than those who had not ( n = 278). Participants were randomized to standard care (SC) or SC plus prize CM. Although prize CM was related to better outcomes overall, recent gambling was not associated with outcomes across or within treatment conditions. Gambling participation before treatment entry was associated with reductions in gambling over time, and this effect was more pronounced among those assigned to CM. These data suggest that prize CM is equally efficacious for substance abusers who do and do not gamble, and they extend prior studies indicating that prize CM does not increase gambling.
Improved methods for expression and purification of S. cerevisiae TFIIF and TFIIH; Identification of a functional E.coli promoter and internal translation initiation within the N-terminal coding region of the TFIIF TFG1 subunit

The basal RNA polymerase II (RNAPII) transcription machinery is composed of RNAPII and the general transcription factors (TF) TATA binding protein (TBP), TFIIB, TFIIE, TFIIF and TFIIH. Due to the powerful genetic and molecular approaches that can be utilized, the budding yeast S. cerevisiae has proven to be an invaluable model system for studies of the mechanisms of RNAPII transcription. Complementary biochemical studies of the S. cerevisiae basal transcription machinery, however, have been hampered by difficulties in the purification of TFIIF and TFIIH, most notably due to the severe toxicity of the TFIIF Tfg1 subunit in E. coli and the complexity of the purification scheme for native TFIIH. Here, we report the elimination of TFG1 -associated toxicity in E. coli , the identification and removal of a functional E.coli promoter and internal translation initiation within the N-terminal coding region of TFG1 , and the efficient production and two step purification of recombinant TFIIF complexes. We also report conditions for the efficient two step tandem affinity purification (TAP) of holo-TFIIH, core TFIIH and TFIIK complexes from yeast whole cell extracts.
Re-examination of the roles of PEP and Mg2+ in the reaction catalysed by the phosphorylated and non-phosphorylated forms of phosphoenolpyruvate carboxylase from leaves of Zea mays

Re-examination of the roles of PEP and Mg2+ in the reaction catalysed by the phosphorylated and non-phosphorylated forms of phosphoenolpyruvate carboxylase from leaves of Zea mays Effects of the activators glucose 6-phosphate and glycine Alejandro TOVAR-ME; NDEZ, Rogelio RODRI;GUEZ-SOTRES, Dulce M. LO; PEZ-VALENTI;N and Rosario A. MUNN OZ-CLARES Departamento de Bioquı! mica, Facultad de Quı! mica, Universidad Nacional Auto! noma de Me! xico, Me! xico D.F. , 04510, Me! xico
Dopamine D 3 Receptor Inactivation Attenuates Cocaine-Induced Conditioned Place Preference in Mice

The dopamine (DA) D3 receptor (D 3 R) has received much attention in medication development for treatment of addiction. However, the functional role of the D 3 R in drug reward and addiction has been a matter of debate. We recently reported that D 3 receptor-knockout (D 3−/− ) mice display increased vulnerability to cocaine self-administration, which we interpret as a compensatory response to attenuated cocaine reward after D 3 R deletion. Here we report that D 3−/− mice displayed attenuated cocaine-induced conditioned place response (CPP) compared to wild-type mice. Similarly, blockade of brain D 3 Rs by YQA-14, a novel DA D 3 receptor antagonist, significantly and dose-dependently inhibits acquisition and expression of cocaine-induced CPP in WT mice, but not in D 3−/− mice. These findings suggest that: 1) D 3 Rs play an important role in mediating cocaine’s rewarding effects; and 2) YQA-14 is a highly potent and selective D 3 R antagonist in vivo, which deserves further study as a candidate for treatment of cocaine addiction. compare CPP response to a commonly used dose of cocaine (20 mg/kg) between WT and D 3−/− mice. We then investigated whether pharmacological blockade of D 3 Rs with YQA-14, a novel highly potent and selective D 3 R antagonist 2012a), alters CPP response to cocaine. YQA-14 shows similar pharmacological properties than SB-277011A. In vitro receptor binding assays suggest that YQA14 has two binding sites on human cloned D3 receptors with K i-High (0.68×10 −4 nM) and K i-Low (2.11 nM), and displays >150-fold selectivity for D3 over D2 receptors and >1000-fold selectivity for D3 over other DA receptors
A simple and efficient method to reduce nontemplated nucleotide addition at the 3 9 terminus of RNAs transcribed by T7 RNA polymerase

DNA templates modified with C2 9 -methoxyls at the last two nucleotides of the 5 9 termini dramatically reduced nontemplated nucleotide addition by the T7 RNA polymerase from both single- and double-stranded DNA templates. This strategy was used to generate several different transcripts. Two of the transcripts were demonstrated by nuclear magnetic resonance spectroscopy to be unaffected in their sequence. Transcripts produced from the modified templates can be purified with greater ease and should be useful in a number of applications.
Inhibiting AP-1 activity alters cocaine induced gene expression and potentiates sensitization

We have expressed A-FOS, an inhibitor of AP-1 DNA binding, in adult mouse striatal neurons. We observe normal behavior including locomotion and exploratory activities. Following a single injection of cocaine, locomotion increased similarly in both the A-FOS expressing and littermate controls. However, following repeated injections of cocaine, the A-FOS expressing mice showed increased locomotion relative to littermate controls, an increase that persisted following a week of withdrawal and subsequent cocaine administration. These results indicate that AP-1 suppresses this behavioral responses to cocaine. We analyzed mRNA from the striatum before and 4 and 24 hours after a single cocaine injection in both A-FOS and control striata using Affymetrix microarrays (430 2.0 Array) to identify genes mis-regulated by A-FOS that may mediate the increased locomotor sensitization to cocaine. A-FOS expression did not change gene expression in the basal state or 4 hours following cocaine treatment relative to controls. However, 24 hours after an acute cocaine treatment, 84 genes were identified that were differentially expressed between the A-FOS and control mice. 56 gene are down regulated while 28 genes are up regulated including previously identified candidates for addiction including BDNF and Per1. Using a random sample of identified genes, quantitative PCR was used to verify the microarray studies. The chromosomal location of these 84 genes was compared to human genome scans of addiction to identify potential genes in humans that are involved in addiction.
A PCR Artifact: Generation of Heteroduplexes

carefully planned for us, and visits to laboratories alternated with visits to the many and well-known attractions of the city. I gave a lecture and took part in a discussion on the prospects of human genetics. The Institute of Medical Genetics is housed in a fourstory building, about half of which is shared with the Endocrinology Institute. It is subdivided into the following departments:
The yeast vacuolar Rab GTPase Ypt7p has an activity beyond membrane recruitment of the HOPS/Class C Vps complex

A previous report described lipid mixing of reconstituted proteoliposomes made using lipid mixtures that mimic the composition of yeast vacuoles. This lipid mixing required SNARE proteins, Sec18p and Sec17p (yeast NSF1 and α-SNAP), and the HOPS/Class C Vps complex, but not the vacuolar Rab GTPase Ypt7p. The present study investigates the activity of Ypt7p in proteoliposome lipid mixing. Ypt7p is required for lipid mixing of proteoliposomes lacking cardiolipin. Omission of other lipids with negatively charged and/or small headgroups does not cause Ypt7p dependence for lipid mixing. Yeast vacuoles made from strains disrupted for cardiolipin synthase (CRD1) fuse to the same extent as vacuoles from strains with functional CRD1. Disruption of CRD1 does not alter dependence on Rab GTPases for vacuole fusion. It has been proposed that HOPS complex recruitment to membranes is the main function of Ypt7p. However, Ypt7p is still required for lipid mixing even when the concentration of HOPS complex in lipid-mixing reactions is adjusted such that cardiolipin-free proteoliposomes with or without Ypt7p bind equal amounts of HOPS. Ypt7p therefore must stimulate membrane fusion by a mechanism in addition to HOPS-membrane recruitment. This is the first demonstration of such a stimulatory activity – that is, beyond bulk effector recruitment – for a Rab GTPase.
A PCR-based method for high stringency screening of DNA libraries

A rapid method for cloning genomic DNA utilizing a PCR-based screening protocol is described. A murine genomic library in lambda phage was subdivided into 64 wells, each containing 1000 clones, and propagated in bacteria. Amplified phage from each of 8 wells across columns, and each of 8 wells down rows, were pooled. The pooled phage were screened for the presence of murine M-CSF DNA by PCR using specific oligonucleotide primers. A single well that contained an M-CSF genomic clone was identified by the synthesis of a PCR product of the correct size that hybridized to an internal M-CSF oligonucleotide probe. This well was subdivided into 64 wells, each containing approximately 30 individual phage, reamplified, and rescreened utilizing the same protocol. A positive well was then subdivided and amplified a third time starting with an average of 2 phage per well, and rescreened for M-CSF DNA by PCR. Phage from a PCR-positive well, now highly enriched for M-CSF DNA, were grown as individual plaques. PCR-screening of randomly picked plaques demonstrated that the majority contained an M-CSF genomic insert. This method obviates the more labor and time intensive method of plaque hybridization screening of DNA libraries, and is more stringent since three oligonucleotides (the two PCR primers, and the hybridization probe) are required to give a true positive signal. Similar methodology has also been used to clone a cDNA gene contained within
A probability score for preoperative prediction of type 2 diabetes remission following RYGB surgery

BACKGROUND— Type 2 diabetes (T2D) is a metabolic disease with significant medical complications. Roux-en-Y gastric bypass (RYGB) surgery is one of the few interventions that remit T2D in ~60% of patients. However, there is no accurate method for predicting preoperatively the probability for T2D remission. METHODS— A retrospective cohort of 2,300 RYGB patients at Geisinger Clinic was used to identify 690 patients with T2D and complete electronic data. Two additional T2D cohorts (N=276, and N=113) were used for replication at 14 months following RYGB. Kaplan-Meier analysis was used in the primary cohort to create survival curves until remission. A Cox proportional hazards model was used to estimate the hazard ratios on T2D remission. FINDINGS— Using 259 preoperative clinical variables, four (use of insulin, age, HbA1c, and type of antidiabetic medication) were sufficient to develop an algorithm that produces a type 2 diabetes remission ( DiaRem ) score over five years. The DiaRem score spans from 0 to 22 and was divided into five groups corresponding to five probability-ranges for T2D remission: 0–2 (88%–99%), 3–7 (64%–88%), 8–12 (23%–49%), 13–17 (11%–33%), 18–22 (2%–16%). The DiaRem scores in the replication cohorts, as well as under various definitions of diabetes remission, conformed to the DiaRem score of the primary cohort.
Sexual dimorphism in development of kidney damage in the aging Fischer-344 (F344) rat

The aging kidney exhibits slowly developing injury and females are usually protected compared to males, in association with maintained renal nitric oxide (NO). Here, we compared renal injury in Fischer 344 (F344) rats in intact, ovariectomized, and ovariectomized + estrogen replaced young (6 month) and old (24 month) female rats with young and old intact male rats and measured renal protein abundance of NO synthase isoforms and oxidative stress. There was no difference in age-dependent glomerular damage between young or old intact male and female F344, and neither ovariectomy nor estrogen replacement affected renal injury; however, tubulointerstitial injury was greater in old males than old females. These data suggest that ovarian hormones do not influence these aspects of kidney aging in F344 rats and that the greater tubulointerstitial injury is caused by male sex. Old males had greater kidney cortex NOS3 abundance than females, and NOS1 abundance (alpha and beta isoforms) was increased in old males compared to both young males and old females. NOS abundance was preserved with age in intact females, ovariectomy did not reduce NOS1 or 3 protein abundance, and estrogen replacement did not uniformly elevate NOS proteins, suggesting that estrogens are not primary regulators of renal NOS abundance in this strain. NADPH oxidase-dependent superoxide production and nitrotyrosine immunoreactivity were increased in the aging male rat kidney compared to the female which could compromise renal NO production and/or bioavailability. In conclusion, the kidney damage expressed in the aging F344 is fairly mild and is not related to loss of renal cortex NOS3 or NOS1 alpha. This is in contrast to the aging male Sprague Dawley rat [1] where kidney damage is exacerbated and NOS3 and NOS1 alpha are lost compared to the old female.
AN ONLINE FORUM ON MENOPAUSAL SYMPTOM EXPERIENCE OF WHITE WOMEN IN THE U.S

Aims—Using a feminist approach, the study aimed at exploring menopausal symptom experience of 23 White midlife women through a six-month online forum. Background—Recent cross-cultural investigations have indicated significant ethnic differences in menopausal symptoms and have challenged the universality of menopausal symptoms. Currently available cultural knowledge on menopausal experience, however, is inadequate to guide appropriate and adequate care even for White midlife women in the menopausal transition. Data source—Qualitative data collected through an online forum in 2007. Method—The study was a cross-sectional qualitative online forum study. A total of 23 midlife women who self-identify as non-Hispanic Whites were recruited for the study using a convenience sampling method. Seven topics related to menopausal symptom experience were used to guide the online forum for 6-months. The data were analyzed using thematic analysis involving line-by-line coding, categorization, and theme extraction. Results—The experience of menopause caused women to redefine themselves within their busy daily life schedules. The women were optimistic about their symptoms, and tried to laugh at the experience to boost their inner strength and motivate themselves persevere. Many women thought that both generational and life styles differences were much more important than ethnic differences in menopausal symptom experience. In seeking assistance with the symptoms of menopause, women were not satisfied with the guidance of their physicians. Conclusion—Nurses need to carefully listen to what the women themselves talked about their own experience with menopausal symptoms and help them to adequately manage and live with the symptoms.
Eects of a range of b2 adrenoceptor agonists on changes in intracellular cyclic AMP and on cyclic AMP driven gene expression in cultured human airway smooth muscle cells

1 The eects of the selective b2 adrenoceptor agonists salbutamol, terbutaline and salmeterol and the non-selective b adrenoceptor agonist isoprenaline on [H]-cyclic AMP formation and cyclic AMP response element (CRE) driven luciferase expression, assessed using the construct p6CRE/luc, were studied in primary cultures of human airway smooth muscle (HASM) cells. 2 Optimal transfection conditions for transient expression of pGL3 Control were 4 mg DNA/well in a 6 well plate and 1.8 ml Transfectam/mg DNA. Expression was maximal at 48 ± 72 h. 3 Salbutamol (maximum response 19%, EC50 0.6 mM), terbutaline (maximum response 38%, EC50 2.3 mM) and salmeterol (maximum response 18%, EC50 0.0012 mM) were all partial agonists for cyclic AMP formation compared with isoprenaline (EC50 0.08 mM). However, all of the b2 adrenoceptor agonists produced increases in CRE-driven luciferase activity, in cultured HASM transfected with the vector p6CRE/luc, which were equivalent or greater (salmeterol) than those seen with isoprenaline. 4 Both salbutamol and salmeterol were more potent at increasing luciferase expression than in elevating cyclic AMP levels in these cells. The potency ratios (EC50 (cyclic AMP)/EC50 (LUC)) for the agents studied were isoprenaline: 0.2 fold, terbutaline: 3 fold, salbutamol: 24 fold, salmeterol: 38 fold. 5 These data suggest that important quantitative dierences exist in the ability of b2 adrenoceptor agonists to increase whole cell cyclic AMP levels in airway smooth muscle and to drive gene expression via a CRE-driven mechanism.