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Mitigation of Obesity: A Phytotherapeutic Approach
Published in Amit Baran Sharangi, K. V. Peter, Medicinal Plants, 2023
A.B. Sharangi, Suddhasuchi Das
Aloe vera belongs to the family Liliaceae. The major chemical composition of A. vera includes phytosterols, anthraquinones, chromones, enzymes, tannins, amino acids, proteins, vitamins, pectins, hemicelluloses, glucomannan, acemannan, and mannose derivatives (Misawa et al., 2012a). A. vera gel powder in Sprague-Dawley rats with diet-induced obesity decreased body weight (Misawa et al., 2012a). Misawa et al. (2012b) reported that feeding of A. vera to Zucker diabetic fatty rats which produces lophenol and cycloartenol (two types of phytosterol), significantly reduced visceral fat weights. Stimulation of energy expenditure is one of the proposed anti-obesity mechanisms of A. vera (Misawa et al., 2012a), other mechanism being the regulation of expression levels of hepatic genes encoding to lipogenic enzymes (ACC, FAS), and transcriptor factor SREBP-1, which, by the administration of aloe sterols, is found to be decreased drastically; and to the increased of hepatic β-oxidation enzymes ACO, CPT1, PPARα (Misawa et al., 2012b).
Non-Hodgkin Lymphoma
Published in Tariq I. Mughal, Precision Haematological Cancer Medicine, 2018
ATLL cells show diverse cytogenetic abnormalities, with the most frequent being gains at chromosomes 14q, 7q and 3p and losses at chromosomes 6q and 13q. The earliest genetic event appears to be epigenome alterations initiated by the HTLV-1 transcriptor of the X-gene region, Tax, through the EZH2 gene. There is then an accumulation of mutations affecting the TCR and NF-κB pathways (PLCG1, PRKCB, CARD11, VAV1 and IRF4), inactivation of tumour suppressors, such as p53, p15INK4A, p16INK4B and mutations in TET2 and MLL3, which are thought to be pivotal in disease development. Other frequently mutated genes are those implicated in cell-cycle genes (CDC2, cyclin B), tyrosine kinase signalling pathways (SYK, LYN), anti-apoptotic factors (BIRC5), calcium metabolism (RANKL, PTHLH), NRXN3, CCR4, CCR7, TSLC1, CAV1 and prostaglandin D2. It is also likely that ATLL cells also affect PD1 receptors and proteins involved in cellular adhesion, within the tumour microenvironment. ENKTL appears to have a unique molecular signature affecting PRDM1, BCOR, ATG5, AIM1 genes and the AURKA, NOTCH-1, NF-κB and JAK/STAT3 pathways.
Multiplex Testing of Bcr-Abl1 and Jak2 V617f in Suspected Mpn Using Rt-Pcr Rdb Method
Published in Cut Adeya Adella, Stem Cell Oncology, 2018
N. Masykura, F. Albertha, A.R.H. Utomo, U. Habibah, M. Yunus, Suharsono, F. Selasih, A. Bowolaksono
RT reverse transcriptase cDNA conversion. Isolated RNAs were converted into cDNA with reverse transcriptase enzyme and random hexamer primers using Transcriptor First Strand cDNA Synthesis (Roche).
Inosine induces acute hyperuricaemia in rhesus monkey (Macaca mulatta) as a potential disease animal model
Published in Pharmaceutical Biology, 2021
Dong-hong Tang, Chen-yun Wang, Xi Huang, Hong-kun Yi, Zhe-li Li, Kai-li Ma, You-song Ye, Jian-wen Zhang
The total RNA was extracted from the liver and kidney biopsies (20 mg) using 1 mL of TriPure (Roche, Basel, Switzerland). RNA integrity was evaluated by 1% agarose electrophoresis, and the samples were stored at −80 °C. Subsequently, 2 μg of RNA was reverse-transcribed using Transcriptor for Strand cDNA Synthesis (Roche, Basel, Switzerland), according to the manufacturer’s protocol. The obtained cDNA was stored at −20 °C. XO (GenBank: NM_000379.3), PNP (GenBank: NM_001193551), URAT1 (GenBank: AB738914.1), GLUT9 (GenBank: NM_020041.2), OAT4 (GenBank: NM_018484) and ABCG2 (GenBank: NM_001032919.1) were amplified by quantitative real-time PCR (qRT-PCR) on a CFX384 Touch™ Real-Time PCR Detection System (Bio-Rad, Hercules, CA). The polymerase chain reaction (PCR) conditions were as follows: initial denaturation for 5 min at 95 °C, and 40 cycles of 5 s at 95 °C and 30 s at 60 °C. Gene expression was quantified by the ΔΔCt method with CXF386 software (Bio-Rad, Hercules, CA), and normalized to GAPDH (GenBank: NM_001195426.1) as an internal control. Primers were designed using Primer 5.0 and synthesized by Takara Biotechnology (Kusatsu, Japan) (Table 1).
Development of model based on clock gene expression of human hair follicle cells to estimate circadian time
Published in Chronobiology International, 2020
Taek Lee, Chul-Hyun Cho, Woon Ryoung Kim, Joung Ho Moon, Soojin Kim, Dongho Geum, Hoh Peter In, Heon-Jeong Lee
One microgram of RNA template from each sample was reverse-transcribed using the Transcriptor First Strand cDNA Synthesis Kit (Roche Diagnostics, Basel, Switzerland) according to the manufacturer’s instructions. For real-time PCR, a 2-μl aliquot of the reverse-transcribed cDNA product was used in each 20-μl sample reaction mixture containing 4 mM MgCl2, 10 pmol of upstream and downstream primer, and 2 μl of 10× LightCycler FastStart DNA Master SYBR Green 1 (Roche Diagnostics, Basel, Switzerland). The sequences of primer sets for the RT-PCR of 10 representative circadian clock genes (PER1, PER2, PER3, CLOCK, ARNTL, CRY1, CRY2, NPAS2, NR1D1, and NR1D2) are shown in Supplementary Table 1.
Modifications in GPR21 and GPR82 genes expression as a consequence of metabolic syndrome etiology
Published in Journal of Receptors and Signal Transduction, 2021
Rodrigo Romero-Nava, Noemí García, Karla Aidee Aguayo-Cerón, Fausto Sánchez Muñoz, Fengyang Huang, Enrique Hong, Santiago Villafaña
After 9 weeks of the administration of fructose or water, rats were sacrificed. The heart, kidney, brain, aorta and blood were removed carefully. Total RNA of the tissues was isolated with Trizol reagent (Life Technologies, Carlsbad, CA) in accordance with the instructions for the manufacturer. Total RNA concentration and purity was quantified using a NanoPhotometer (Implen, inc. Westlake Village, CA) measurement of the optical densities at 260/280 nm and 260/230. A ratio of 1.8–2.2 A260/A280 and A260/A230 was required for these studies. The cDNA was prepared using Transcriptor First Strand cDNA Synthesis Kit Reverse Transcriptase (Roche Diagnostics, Mannheim, Germany). Approximately 1 µg of total RNA was reverse transcribed and used for real-time PCR analysis using the Nano LightCycler System (Roche Diagnostics, Mannheim, Germany) in a total reaction mixture volume of 10 µL containing 5 µL FastStart Essential DNA Probes Master (Roche Applied Science, Mannheim, Germany), 0.15 µL of probe Universal ProbeLibrary (Roche Applied Science, Mannheim, Germany), 0.3 µL left primer (Sigma-Genosys Ltd, UK), 0.3 µL right primer (Sigma-Genosys Ltd, UK), 3.25 µL water PCR grade and 1 µL sample. Comparative cycle values were used to determine expression levels. The genes expression was normalized by β-actin expression levels. PCR primers was as follow: GPR21 receptor, 5′-GAACTCCACCTGGGATGGTA-3′ (forward) and 5′-GTAGCCCAGTGCCAGAAGAC-3′ (reverse); GPR82 receptor, 5′-CCAGTTCTCCAAGACAGAGCTT-3′ (forward) and 5′-GATTGATCCTGAGAAGCTGGATAA-3′ (reverse); β-actin, 5′-CCCGCGAGTACAACCTTCT-3′ (forward) and 5′-CGTCATCCATGGCGAACT-3′ (reverse). The GPR21 and GPR82 receptors mRNA expression levels were determined by the comparative 2-ΔΔCt method.