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#25477280   2015/05/01 To Up

The methyltransferases enhancer of zeste homolog (EZH) 1 and EZH2 control hepatocyte homeostasis and regeneration.

To investigate the role of enhancer of zeste homolog (EZH) 1 and EZH2 in liver homeostasis, mice were generated that carried Ezh1(-/-) and EZH2(fl/fl) alleles and an Alb-Cre transgene. Only the combined loss of EZH1 and EZH2 in mouse hepatocytes caused a depletion of global trimethylation on Lys 27 of histone H3 (H3K27me3) marks and the specific loss of over ∼1900 genes at 3 mo of age. Ezh1(-/-),Ezh2(fl/fl)Alb-Cre mice exhibited progressive liver abnormalities manifested by the development of regenerative nodules and concomitant periportal fibrosis, inflammatory infiltration, and activation of A6-positive hepatic progenitor cells at 8 mo of age. In response to chronic treatment with carbon tetrachloride, all experimental mice, but none of the controls (n = 27 each), showed increased hepatic degeneration associated with liver dysfunction and reduced ability to proliferate. After two-thirds partial hepatectomy, mutant mice (n = 5) displayed increased liver injury and a blunted regenerative response. Genome-wide analyses at 3 mo of age identified 51 genes that had lost H3K27me3 marks, and their expression was significantly increased. These genes were involved in regulation of cell survival, fibrosis, and proliferation. H3K27me3 levels and liver physiology were unaffected in mice lacking either EZH1 globally or EZH2 specifically in hepatocytes. This work demonstrates a critical redundancy of EZH1 and EZH2 in maintaining hepatic homeostasis and regeneration.

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EZH2 KMT6 Control Peptid EZH2 KMT6 Control Peptid Anti-PCOLE-1 (Procollagen Anti PCOLE 1 (Procollagen anti-Hairy and Enhancer o Anti blastemal regenerati Anti-blastemal regenerati EZH2 GSK-126 Mechanisms: EZH2 Methyltransferase EZH2 As EZH2 KMT6 antibody Isoty Rabbit Anti-KMT6 EZH2 Pol

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#28248099   2017/03/01 To Up

Efficient Expression of an Acidic Endo-polygalacturonase from Aspergillus niger and Its Application in Juice Production.

An endo-polygalacturonase gene (pga-zj5a) was cloned by reverse transcription from cDNAs synthesized from Aspergillus niger ZJ5 total RNA. The open reading frame of pga-zj5a was 1089 base pairs encoding 362 amino acids. Pga-zj5a lacking a signal peptide sequence was successfully amplified using A. niger ZJ5 cDNA as the template and was ligated into the pPIC9 vector. The resulting plasmid was transformed into competent cells of Pichia pastoris GS115 for heterologous expression. The polygalacturonase showed a maximum activity level of 10436 U/mL in the culture supernatant from a 3 L fermenter. Assays of enzymatic properties showed that the optimal pH and temperature of the recombinant PGA-ZJ5A were 4.5 and 40 °C, respectively. PGA-ZJ5A was effective in pear juice clarification, increased the volume of pear juice by 41.8%, and improved its light transmittance 3-fold.

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pNosdcGUS Plant GUS Expre Mouse Fibroblast Growth F Human Fibroblast Growth F MarkerGeneTM Biotin X Ant Custom Recombinant Protei Custom Monoclonal Antibod Custom Polyclonal Antibod ENDO-Growth Medium ENDO-Basal Medium ENDO-Growth Supplement ENDO AGAR BASE Mouse Anti-Aspergillus sp

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#28447263   2017/04/27 To Up

Design of a Lentiviral Vector for the Inducible Expression of MYC: A New Strategy for Construction Approach.

Lentiviral vectors are powerful tools for gene expression studies. Here we report the construction of pTIJ, a vector for inducible gene expression. pTIJ was generated from pTRIPZ backbone, which is designed for the inducible expression of shRNA sequences, by the introducing of a multiple cloning site upstream of the Tet promoter and the removal of miR30 flanking sequences. To evaluate pTIJ as a tool for the inducible expression of genes of interest, we introduced MYC cDNA into pTIJ and infected two small cell lung cancer cell lines, H209 and H345. Induction of MYC expression by doxycycline was detectable in both cell lines by real-time PCR and western blot analysis. This study highlights the relevance of pTIJ vector to allow the inducible expression of any gene of interest. In our belief, pTIJ will be an extremely useful tool to simplify the generation of genetically engineered cell lines for the inducible expression of cDNA sequences in biological studies. Furthermore, we report the generation of a pTIJ-MYC vector for the inducible expression of the oncogene MYC.

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Lentiviral Technology pCD pNosdcGUS Plant GUS Expre pCAMBIA0105.1R Vector, (G pDC57 Mammalian Luciferas pDC99 Mammalian Luciferas pCAMBIA1200 Vector (No Re pCMVLuxB Mammalian LuxB E pCMVLuxA Mammalian LuxA E pCAMBIA1201 Vector (gusA pCAMBIA1281Z Vector (gusA pCAMBIA1305.1 Vector (Gus pCAMBIA2200 Vector (No Re

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#28708518   2017/07/14 To Up

Ultrasonography to assist with timing of spawning in European eel.

Ovulation in the European eel can be induced by injection of DHP (17α,20β-dihydroxy-4-pregnen-3-one). The timing of injection is based on the developmental stage of the oocytes. The oocyte stage is determined by invasive biopsy or through external indicators of the oocyte hydration response: body weight index (BWI) and body girth index (BGI). However, in European eel, BWI and BGI are inaccurate indicators because the individual hydration response is highly variable. The aim of this study was to identify indicators of oocyte maturation non-invasively by applying ultrasonography. Ultrasound parameters that were determined were the body wall thickness; diameter of the cloaca opening; grey scale median (GSM) of the ovary, and diameters (vertical and horizontal) and surface area of ovary, liver and swim bladder. Data of farmed and wild eels were combined for correlation analyses and principal component analyses (PCA). Ovary and liver parameters were not correlated with the average oocyte stage. Body wall thickness was negatively correlated to the average oocyte stage but variability was high. The swim bladder vertical diameter (Sbv) was negatively correlated with the average oocyte stage and with the oocyte diameter. Swim bladder area (Sba) was also negatively correlated to the average oocyte stage but less accurate. Results showed that during the final stages of oocyte maturation, the swim bladder becomes smaller. This is most probably due to the pressure that is caused by the increasing ovaries as a result of the oocyte hydration response. Females with an average oocyte stage between 4.5 and 7.0 and an average oocyte diameter higher than 800 μm, had Sbv values decreasing from 4.3 to 2.4 mm and Sba values from 52 down to 11 mm(2). The correlation between Sbv and oocyte stage is more strict than for BWI or BGI which makes Sbv not only a parameter that can be determined non-invasively but also one that is more accurate in indicating the oocyte developmental stage. Therefore, ultrasonography, and in particular the ultrasound imaging of the swim bladder, represents a useful tool to assist with the timing of spawning as induced by injection of DHP.

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#28708470   2017/07/14 To Up

Safety and Efficacy of an Absorbable Filter in the Inferior Vena Cava to Prevent Pulmonary Embolism in Swine.

Purpose To evaluate the immediate and long-term safety as well as thrombus-capturing efficacy for 5 weeks after implantation of an absorbable inferior vena cava (IVC) filter in a swine model. Materials and Methods This study was approved by the institutional animal care and use committee. Eleven absorbable IVC filters made from polydioxanone suture were deployed via a catheter in the IVC of 11 swine. Filters remained in situ for 2 weeks (n = 2), 5 weeks (n = 2), 12 weeks (n = 2), 24 weeks (n = 2), and 32 weeks (n = 3). Autologous thrombus was administered from below the filter in seven swine from 0 to 35 days after filter placement. Fluoroscopy and computed tomography follow-up was performed after filter deployment from weeks 1-6 (weekly), weeks 7-20 (biweekly), and weeks 21-32 (monthly). The infrarenal IVC, lungs, heart, liver, kidneys, and spleen were harvested at necropsy. Continuous variables were evaluated with a Student t test. Results There was no evidence of IVC thrombosis, device migration, caval penetration, or pulmonary embolism. Gross pathologic analysis showed gradual device resorption until 32 weeks after deployment. Histologic assessment demonstrated neointimal hyperplasia around the IVC filter within 2 weeks after IVC filter deployment with residual microscopic fragments of polydioxanone suture within the caval wall at 32 weeks. Each iatrogenic-administered thrombus was successfully captured by the filter until resorbed (range, 1-4 weeks). Conclusion An absorbable IVC filter can be safely deployed in swine and resorbs gradually over the 32-week testing period. The device is effective for the prevention of pulmonary embolism for at least 5 weeks after placement in swine. (©) RSNA, 2017.

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Lung disease spectrum (pu Lung disease spectrum (pu Homogenizer for 24 sample Celite@ 500 Filter Aid Stat3 Filter Assay Tissue Filter Tubes, 100 MCE Membrane Filter, Pore Circulaire FB250 Filter B Transcription Factors: T Transcription Factors: N MCE Gridded Membrane Filt MCE Gridded Membrane Filt

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#28708532   2017/07/14 To Up

Blood flow and echotextural differences between the future dominant and subordinate follicles before the beginning of diameter deviation in heifers.

Diameter deviation is the beginning of a decrease in growth rate of the largest subordinate follicle (SF) and a continuing growth rate of the dominant follicle (DF). In wave 1 in cattle, deviation begins 2 or 3 days after ovulation when the future DF is about 8.5 mm. Gray scale and power-flow Doppler ultrasound examinations were done in experiment 1 (daily examinations, n = 13) and experiment 2 (examinations every 8 h, n = 15). Blood flow and an anechoic layer in the follicle wall were normalized to the beginning of diameter deviation (day 0 or hour 0). Only waves with conventional diameter deviation (68% of waves) were used as identified by: (1) future SF greater than 7.0 mm when DF was 8.5 mm and (2) future DF and SF did not switch in diameter rank. In experiment 1, deviations in the extent of blood-flow signals and in the extent of anechoic layer began on the same day as deviation in diameter. In experiment 2, deviations in diameter, surface area (πd(2)), and anechoic layer began in synchrony, and deviation in blood-flow signals began 16 h earlier. Blood-flow deviation before diameter deviation was shown by (1) a first difference (P < 0.02) between follicles at hour -16 and (2) development during the hours -24 to 0 of a greater (P < 0.05) percentage difference between follicles in blood-flow signals (11.1 ± 2.3%) than in surface area (7.4 ± 0.7%) or diameter (4.5 ± 0.4%). Results supported the hypothesis that the extent of blood flow in the future dominant and subordinate follicles deviates before diameter deviates. A similar hypothesis for anechoic layer was not supported; diameter and anechoic layer deviated in synchrony.

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#28708540   2017/07/14 To Up

Influence of two ovulation-inducing agents on the pituitary response and follicle blood flow in mares.

The objective of the current study was to evaluate the effects of deslorelin and hCG, two ovulation-inducing therapies, on LH surge and follicle vascularity in mares. Thirty mares were either treated with 1.5 mg IM of deslorelin, 2,500 IU IV of hCG or 2 mL IM of NaCl 0.9% (GnRH, hCG and Saline groups, respectively). Power-flow Doppler examination and blood collection were performed every hour during the first 12 hours after treatment (H0) and every six hours between hours 12 (H12) and 30 (H30) after treatment. Moreover, endpoints were evaluated every hour through the last six hours before ovulation (OV-6 to OV-1). In GnRH group, plasma LH concentration progressively increased (P < 0.001) during the first 6 hours after treatment and remained high (P > 0.1) until OV-1. A significant increase in LH concentrations was first detected (P < 0.05) at 24 hours after treatment in hCG group, while no changes (P > 0.1) on LH levels were found during H0-H30 and between OV-6 and OV-1 in the Saline group. Independent of the treatment, significant variations on the percentage of the follicle wall with Doppler signals were not observed (P > 0.1) throughout the entire experiment. A weak correlation between the preovulatory follicle vascularity and the plasma LH concentration was found in GnRH, hCG and Saline groups (r = +0.29, +0.29 and -0.23, respectively; P ˂ 0.0001). These results described for the first time the immediate and continuous pituitary response to ovulation-inducing therapy with injectable deslorelin. Moreover, spontaneous and induced ovulations were not preceded by an increased follicle vascularity, which differs from previous reports in large animals.

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#28708616   2017/07/14 To Up

Monitoring accumulation sediment characteristics in full scale sewer physical model with urban wastewater.

A series of experiments were carried out with real wastewater in a pilot flume located at A Coruña wastewater treatment plant (WWTP) (Spain). A full scale model was developed to test a circular (300 mm inner diameter) and an equivalent area egg-shaped plastic pipe under controlled experimental conditions (pipe slope 2-5‰, averaged discharge Q = 4 L/s). Velocity profiles and sediment accumulation in the pipe invert was daily measured. Within the 7-11 days, the average sediment accumulation rate found in the circular pipe was between 1.4 and 3.8 mm/d. The sediment height depended on the input wastewater sediment distribution and organic content. The egg-shaped pipe presented no sediment deposit for the same downstream boundary conditions, although biofilms were attached to the walls of both pipes. Besides, wastewater quality was monitored continuously and sediment composition was studied at the end of experiments. Two types of sediment were recorded: a granular bed deposit (ρ = 1,460 kg/m(3), d50 = 202 μm) and wall biofilms (ρ = 1,190 kg/m(3), d50 = 76 μm).

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ChromaLink™ Digoxigenin 5 (Octadecanoylamino)fluo 4 Methylumbelliferyl sulf Model 150 V T Ultrasonic Model 300 V T Ultrasonic ReadiLink™ mFluor™ Vi ReadiLink™ mFluor™ Vi Advanced Airway Intubatio Model 3000 Ultrasonic Hom MarkerGene™ LysoLive™ Diaph1 Full Length SIVgag251 (full length)

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#28708775   2017/07/14 To Up

Continuous Cardiac Autonomic and Hemodynamic Responses to Isometric Exercise.

Elevated arterial blood pressure (BP) is associated with autonomic dysfunction and impaired hemodynamic control mechanisms. Isometric exercise (IE) training has been demonstrated effective at reducing BP; however, the continuous cardiovascular responses during IE are underinvestigated. We hypothesized that reflex autonomic cardiovascular control is an important mediator in reducing BP. To test our hypothesis, we investigated continuous cardiac autonomic modulation and baroreceptor reflex sensitivity (BRS) in response to IE.

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MCE Gridded Membrane Filt MCE Gridded Membrane Filt Primary antibody DRAK1 A Primary antibody DRAK2 A Human Cardiac Troponin-T Human Cardiac Troponin-T Human Cardiac Troponin-T Pig Cardiac FABP ELISA Ki Recombinant Human Cardiac Recombinant Human Cardiac Recombinant Human Cardiac Recombinant Human Cardiac

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#28708841   2017/07/14 To Up

NlpD links cell wall remodeling and outer membrane invagination during cytokinesis in Escherichia coli.

Cytokinesis in gram-negative bacteria requires the constriction of all three cell envelope layers: the inner membrane (IM), the peptidoglycan (PG) cell wall and the outer membrane (OM). In order to avoid potentially lethal breaches in cell integrity, this dramatic reshaping of the cell surface requires tight coordination of the different envelope remodeling activities of the cytokinetic ring. However, the mechanisms responsible for this coordination remain poorly defined. One of the few characterized regulatory points in the envelope remodeling process is the activation of cell wall hydrolytic enzymes called amidases. These enzymes split cell wall material shared by developing daughter cells to facilitate their eventual separation. In Escherichia coli, amidase activity requires stimulation by one of two partially redundant activators: EnvC, which is associated with the IM, and NlpD, a lipoprotein anchored in the OM. Here, we investigate the regulation of amidase activation by NlpD. Structure-function analysis revealed that the OM localization of NlpD is critical for regulating its amidase activation activity. To identify additional factors involved in the NlpD cell separation pathway, we also developed a genetic screen using a flow cytometry-based enrichment procedure. This strategy allowed us to isolate mutants that form long chains of unseparated cells specifically when the redundant EnvC pathway is inactivated. The screen implicated the Tol-Pal system and YraP in NlpD activation. The Tol-Pal system is thought to promote OM invagination at the division site. YraP is a conserved protein of unknown function that we have identified as a new OM-localized component of the cytokinetic ring. Overall, our results support a model in which OM and PG remodeling events at the division site are coordinated in part through the coupling of NlpD activation with OM invagination.

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