isolation reverse transcription and quantitative real time pcr

... spectrophotometrically and μg of RNA was reversed transcripted using cDNA synthesis kit (Bio-Rad) and Real- time PCR and quantitative PCR were performed to assess expression of the COX-2, LPA1, LPA2, LPA3, LPA4, and ... Methods and were cultured in 6-well plates A) Total RNA was isolated and LPA receptors mRNA levels were measured by Real- time RT -PCR B) Cells were challenged with 18:1LPA (1 μM) for h, and COX-2 ... Welfare Act and the National Institutes of Health guidelines for the care and use of animals in biomedical research PCR genotyping of LPA1+/- and LPA2+/- mice Extract-N-Amp Tissue PCR kit (Sigma...

Ngày tải lên: 12/08/2014, 14:20

... respectively 1.9, 5.1, 2.3, 1.8, and 2.6 times compared to fed ND mice While, concentration of HDL-c and enzymatic activity of lipase decreases respectively 1.6 times and 2.2 times as compared to ND fed ... obesity and, possibly diabetes [2] Anti-hyperlipidemic and anti-diabetic effect of metformin were showed, but side-effect and its efficacy of remains in debate if its use for a long time It is ... enzymatic activity of the digestion and absorption of dietary fat such as lipase, lipoprotein lipase and enhancement of carnitine acyl transferases activity and fatty acid oxidation enzymes in...

Ngày tải lên: 14/03/2014, 10:20

... were not even given names and Steinbeck's dislike for them is obvious This fact truly illustrates the message he is trying to get across In "Of mice and Men" the boss and his son Curley are portrayed ... the better.2.) Description of Ma and the preacher, how they were characterized out of their traditional roles and how their straying form the norm was justified and helped relay to the reader the ... Lennie and George, how they were outcasts and Lennie killed a women yet the reader felt sorry for them both because they were on the opposite side of a greater injustice.4.) Portrait of the rich and...

Ngày tải lên: 21/03/2014, 22:03

... WBC number on day and 11 (80% and 88% respectively) in mice Recovery of WBC count started on day and day 11 in all treatment groups and returned back to the normal level on day and day 14 in NS ... significantly increased the number of blood vessel and PCNA-positive cell in both the gastric (Fig 2A and 2B respectively) and duodenal tissues (Fig 3A and 3B respectively) Dosedependent effects were ... Apigenin inhibits VEGF and HIF-1 expression via PI3K/AKT/p70S6K1 and HDM2/p53 pathways FASEB J 2005; 19: 342-53 28 Moallem SA, Hales BF The role of p53 and cell death by apoptosis and necrosis in 4-hydroperoxycyclophosphamide-induced...

Ngày tải lên: 02/11/2012, 10:19

... handoff of the host controller The time line in Figure starts with a power-up (cold boot) event on the PC 8* Immediately after power-up and for some period of time, the BIOS controls the PC and ... first host controller command Read current value of USB COMMAND REGISTER Clear RS bit in the USB COMMAND REGISTER // Will stop host controller Clear CF bit in the USB COMMAND REGISTER // Will signal ... sends Global Reset signal on the GRESET USB and then resets all its logic = host controller resets its internal timers, counters, HCRESET state machines, and so on to their initial values = Run (host...

Ngày tải lên: 07/10/2013, 00:20

... insulin and key hepatic enzymes in experimental diabetes, Pharmaceutical Biology 40(3), (2002) 165 [8] N Brandstrup, JE Kirk, C Bruni, The hexokinase and phosphoglucoisomerase activities of aortic and ... study is to investigate hypolipidemic and hypoglycaemic effects of extract fractions from Ipomoea batatas leaves Materials and methods 2.1 Plant material and preparation of sweet potato leaf extract ... water, dried at 500C and grinded into powder Powdered samples were extracted repeatedly three times with 10 volumes of 90% ethanol by continuous stirring The extract was filtered and lyophilized to...

Ngày tải lên: 12/02/2014, 17:20

... this mutant strain as described below) Body and organ weights as well as plasma parameters (Tables S2 and S3) were measured, and learning, memory and behavioral tests (Table S4) as well as histopathological ... mice (*P < 0.05, Student’s t test) E2F1, E2F transcription factor 1; E2F4, E2F transcription factor 4; E2F5, E2F transcription factor 5; E2F8, E2F transcription factor 8; MKP-1, MAP kinase phosphatase1; ... Matsushime, Masato Kobori, Jiro Hirosumi and Mr Tsutomu Higashiya at Astellas Pharma Inc., and Junko Kawano and Akemi Matsuoka at Trans Genic Inc for their helpful advice and support 12 References 13 Zhang...

Ngày tải lên: 16/02/2014, 09:20

... fed with standard diet and drinking water ad libitum + Lot 2: Obese mice treated with ethanol fraction and drinking water ad libitum + Lot 3: Obese mice treated with chloroform fraction and drinking ... fraction and drinking water ad libitum + Lot 5: Obese mice treated with metformin (500mg/kg) and drinking water ad libitum + Lot 6: Obese mice fed normal diet and drink water (no treatment) and drinking ... consist of phenolic, glycoside and alkaloid compounds However, phenolic compounds as flavonoids, tannin, catechin and alkaloid were found principally in ethanol, n-hexan and chloroform fractions Especially,...

Ngày tải lên: 05/03/2014, 11:21

... Science, Natural Sciences and Technology 27 (2011) 125-133 Introduction Obesity can be considered as an abnormal state of lipid metabolism and increasing in developed and even developing countries ... remedies were limited [6] For a long time, the Docynia indica (W.) Decne fruits were used to prepare traditional remedies for treatment of in and infectious diseases and digestive, but its anti-obesity ... National Institute of Hygiene and Epidemiology (NIHE) were housed in a temperature (25 ± 20C) and humidity (80% ± 10 %) under a 12h light/12 h dark cycle Diets and tap water were provided ad...

Ngày tải lên: 05/03/2014, 11:21

... Bachem AG (Bubendorf, Switzerland) Fetal bovine serum, RPMI-1640, penicillin and streptomycin were purchased from Gibco Laboratories (Grand Island, NY, USA) Caspase-3 and PARP1 antibodies were purchased ... Nordrehaug JE, Ueland M & Kvale G (1995) Total plasma homocysteine and cardiovascular risk profile The Hordaland Homocysteine Study JAMA 274, 1526–1533 11 Nygard O, Refsum H, Ueland PM, Stensvold ... incubation time of h, resulted in reduction of caspase-3 activity (Fig 1C) and apoptosis (Fig 1D) Through reduction of the apoptotic process, the levels of cleaved PARP1, a caspase-3 substrate, and...

Ngày tải lên: 07/03/2014, 09:20

... between and 1, to compare their variation and the midpoint transition (Fig 4) The ensemble of data provides evidence of a common x0,mid between and Time- resolved fluorescence Fluorescence lifetime ... ‡ 10 and reaching Ó FEBS 2004 Fig Fluorescence lifetimes and fraction of the short-lived component Fluorescence lifetimes, s1 (j) and s2 (m), and fraction of the shortlived component, f1 (s), ... for absorption at the excitation wavelength) and Fo and Fw are, respectively, the fluorescence intensities in the absence and Ó FEBS 2004 736 S M Andrade et al (Eur J Biochem 271) presence of...

Ngày tải lên: 07/03/2014, 15:20

... donor (TLL tryptophans) and an acceptor (fatty acid) is that there must exist a spectral overlap between the donor emission and the acceptor absorption spectra, and the donor and acceptor groups ... crystallization and crystallography TLL(S146A) solution was washed several times in 10K Centricon in 10 mM Tris/HCl pH 8.0 buffer and concen- Ó FEBS 2002 Lipase binding to lipid, FRET and structural ... concentration Data from Fig Fig FRET between TLL (and its mutants) and cis-PnA in the presence and absence of NaTDC The protein concentration was 0.8 lM and the cis-PnA concentration was varied stepwise...

Ngày tải lên: 08/03/2014, 23:20

... medium, and cultured for 3, and 14 days RT -PCR Total RNA from cell culture samples was isolated with the RNeasy Micro Kit (Qiagen) and subjected to RT -PCR with the Superscript II kit (Invitrogen) PCR ... WT and TTR KO BMSCs as assessed by ALP, osteocalcin and osteopontin levels (A) ALP activity of WT and TTR KO BMSCs under osteoblast differentiation conditions at days 3, and 14 (B–D) Semiquantitative ... of WT and TTR KO mice (3 months old) Scale bars: 50 lm B C A C 268 B Fig NPY and PAM expression in bone cells from WT and TTR KO mice (A) NPY RT -PCR analysis in brain, MC3T3-E1 cells, and BMSCs...

Ngày tải lên: 15/03/2014, 09:20

... (lanes and 3), CD69NG70 (lanes and 5), CD69NV82 (lanes and 7) CD69NS84 (lanes and 9), rat CD69 (lanes 10 and 11) and mouse CD69 (lanes 12 and 13) was performed under nonreducing (even lanes) and ... 5¢-CTCGAGACAATACAATTGTCCAGG-3¢, and reverse primer 5¢-ACAAAGCTTATTTGTAAGGTTTGTT ACA-3¢, and the PCR product was cloned into pBSK+ vector using the SmaI restriction site, and then into pRSETB vector using XhoI and HindIII ... solubility and stability among these proteins and allow us to select the human protein containing amino acids Gly70 to Lys199 (and the rat and mouse orthologs) as the most physically and biochemically...

Ngày tải lên: 16/03/2014, 04:20

... (Pierce) of the protected probe and RNA ladder Quantitative RT -PCR Quantitative reverse transcription- PCR (qRT -PCR) was used to determine the number of copies of mature and unspliced CB1 transcripts ... (M-MLV) reverse transcriptase and random decamers were used to synthesize single-stranded cDNA 5¢-RLM-RACE PCR was performed using an outer adapter primer (5¢-GCTG ATGGCGATGAATGAACACTG-3¢) and MMCB1 ... wild-type and HD transgenic mice Striatal RNA was extracted from 3-, 5-, 6-, and 12 week-old wildtype and R6/1 transgenic HD mice (n ¼ per age and genotype) Three gene-specific primers and M-MLV reverse...

Ngày tải lên: 16/03/2014, 18:20

... disrupted promoter structure and exons I and II, exhibit impaired follicular development and ovulation [25] and impaired spermiogenesis in males [26] On the other hand, mice defective in Camk4 ... hemisphere and between cerebellum primordium and medulla oblongata (Fig 6, A1 and A2) Also border regions between metencephalon and pons with the third ventricle (arrows in Fig 6, A1 and A2) showed ... using the standard primers T7 and SP6 All PCR products were purified, checked by electrophoresis, quantified and sequenced before use as probes for hybridization Northern blot Total RNA isolation...

Ngày tải lên: 16/03/2014, 23:20

... humans and S pombe (A) Q10 standard, (B) wild-type (WT), (C) E coli harboring pBSDPS1 and pSTVhDLP1, (D) pBmSPS1 and pSTVhDLP1, (E) pBSDPS1 and pSTVmDLP1, (F) pUhDPS1 and pSTVmDLP1, (G) pBmSPS1 and ... kDa), myoglobin (17 kDa) and vitamin B12 (1.35 kDa) standards (Lower) Fractions containing standards and analyzed proteins of (1) GST–mSPS1 and (2) His–mDLP1, or (3) GST–hDPS1 and (4) His–hDLP1, or ... mSPS1 and hDPS1 were 1230 and 1245 bp, respectively, whereas those of mDLP1 and hDLP1 were 1206 and 1200 bp, respectively The mSPS1 and hDPS1 genes were 83.0% identical, and their translated products...

Ngày tải lên: 16/03/2014, 23:20

... weight and fasting plasma cholesterol and triglyceride levels were determined at the indicated time- points after diet initiation At the end of each experiment, mice were killed, and plasma and liver ... 0.005) To determine plasma insulin and leptin levels in these mice, serum samples were isolated at weeks and 24 of the experiment and analyzed for insulin and leptin At week 0, all three mouse ... 60 75 Time (min) 45 60 75 90 105 120 Time (min) 90 105 120 Glucose levels (mg·dL–1) Fig Glucose tolerance curves (A, B) and insulin sensitivity curves (C, D) of ApoE3knock-in, C57BL/6 and ApoE)/)...

Ngày tải lên: 17/03/2014, 17:20

... also just as lonely as the farm workers, is bored and comes in and begins to harass him, and he realizes that he really has no hope of these things really happening The next day, Lennie receive's ... generally friendly, and let he and Candy in They talked about their dream of their own place, and Crooksdecided he'd like to join them He said he'd work for his keep They all agreed on it, and nowCrooks ... the old dog had been killed, Candy realized that he to would soon be old and unwanted like the dog was, so, wanting to have companions, he decided to join Lennie and George in their dream of having...

Ngày tải lên: 21/03/2014, 22:48

Ngày tải lên: 21/03/2014, 22:48