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https://www.readbyqxmd.com/read/27932754/-overexpression-of-lentivirus-rfx1-and-its-inhibitory-effect-on-proliferation-of-glioblastoma-cells
#1
Kai Cheng, Haojie Sun, Mingzhi Zhang, Li Shen
To construct overexpression lentivirus vector for human regulatory factor X1 (RFX1) gene, and to explore its effect on proliferation of F98 cell line.
 Methods: Huamn RFX1 gene was amplified by polymerase reaction. Gene amplification products were inserted into lentivirus vector pITA, and the lentivirus vector pITA-RFX1 was constructed. The constructed vector was verified by agarose gel electrophoresis and DNA sequencing. Lentivirus vector pITA-RFX1 and virus packaging plasmids were cotransfected into 293T cells, and then transfected into F98 cells...
November 28, 2016: Zhong Nan da Xue Xue Bao. Yi Xue Ban, Journal of Central South University. Medical Sciences
https://www.readbyqxmd.com/read/27925002/a-novel-adjuvant-solution-layer-strategy-for-improving-the-efficacy-of-cryosurgery
#2
K K Ramajayam, A Kumar, S K Sarangi, A Thirugnanam
BACKGROUND: During cryosurgery, studies reported earlier suggest increased destruction inside the tumour due to adjuvants or the prevention of damage to neighbouring healthy tissue through different methods. OBJECTIVE: This study advocates a novel strategy that increases the freezing inside agarose gel phantoms by addition of glycine and limits the freezing to the desired location using a perfluorohexane layer during cryosurgery. MATERIALS AND METHODS: Cryosurgery of glycine containing gels is carried out with and without perfluorohexane layer and the thermal history is measured using K-type thermocouples connected to a data acquisition system...
September 2016: Cryo Letters
https://www.readbyqxmd.com/read/27915178/anticancer-copper-pyridine-benzimidazole-complexes-ros-generation-biomolecule-interactions-and-cytotoxicity
#3
Kathleen E Prosser, Stephanie W Chang, Felix Saraci, Phuc H Le, Charles J Walsby
The Cu(II) complex CuCl2(pbzH), pbzH=2-(2-pyridyl)benzimidazole, and derivatives modified at the non-coordinated nitrogen of the benzimidazole fragment, have been studied as anticancer agents. These compounds show promising cytotoxicity against A549 adenocarcinomic alveolar basal epithelial cells with IC50 values in the range of 5-10μM. Importantly, this activity is higher than either CuCl2·2H2O or the individual ligands, demonstrating that ligand coordination to the Cu(II) centres of the complexes is required for full activity...
November 5, 2016: Journal of Inorganic Biochemistry
https://www.readbyqxmd.com/read/27909922/production-of-biologically-active-recombinant-goose-fsh-in-a-single-chain-form-with-a-ctp-linker-sequence
#4
Hui Li, Huanxi Zhu, Qinming Qin, Mingming Lei, Zhendan Shi
FSH is a glycoprotein hormone secreted by the pituitary gland that is essential for gonadal development and reproductive function. In avian reproduction study, especially in avian reproduction hormone study, it is hindered by the lack of biologically active FSH. In order to overcome this shortcoming, we prepared recombinant goose FSH as a single chain molecule and tested its biological activities in the present study. Coding sequences for mature peptides of goose FSH α and β subunits were amplified from goose pituitary cDNA...
December 1, 2016: Molecular Biology Reports
https://www.readbyqxmd.com/read/27908180/a-novel-approach-to-evaluate-spatial-resolution-of-mri-clinical-images-for-optimization-and-standardization-of-breast-screening-protocols
#5
Marco Borri, Erica D Scurr, Cheryl Richardson, Marianne Usher, Martin O Leach, Maria A Schmidt
PURPOSE: Stringent quality assurance is required in MRI breast screening to ensure that different scanners and imaging protocols reach similar diagnostic performance. The authors propose a methodology, based on power spectrum analysis (PSA), to evaluate spatial resolution in clinical images. To demonstrate this approach, the authors have retrospectively compared two MRI sequences commonly employed in breast screening. METHODS: In a novel approach to PSA, spatial frequency response curves (SFRCs) were extracted from the images...
December 2016: Medical Physics
https://www.readbyqxmd.com/read/27897195/arrangement-at-the-nanoscale-effect-on-magnetic-particle-hyperthermia
#6
E Myrovali, N Maniotis, A Makridis, A Terzopoulou, V Ntomprougkidis, K Simeonidis, D Sakellari, O Kalogirou, T Samaras, R Salikhov, M Spasova, M Farle, U Wiedwald, M Angelakeris
In this work, we present the arrangement of Fe3O4 magnetic nanoparticles into 3D linear chains and its effect on magnetic particle hyperthermia efficiency. The alignment has been performed under a 40 mT magnetic field in an agarose gel matrix. Two different sizes of magnetite nanoparticles, 10 and 40 nm, have been examined, exhibiting room temperature superparamagnetic and ferromagnetic behavior, in terms of DC magnetic field, respectively. The chain formation is experimentally visualized by scanning electron microscopy images...
November 29, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27896772/characterization-of-a-protein-interactome-by-co-immunoprecipitation-and-shotgun-mass-spectrometry
#7
Giuseppina Maccarrone, Juan Jose Bonfiglio, Susana Silberstein, Christoph W Turck, Daniel Martins-de-Souza
Identifying the partners of a given protein (the interactome) may provide leads about the protein's function and the molecular mechanisms in which it is involved. One of the alternative strategies used to characterize protein interactomes consists of co-immunoprecipitation (co-IP) followed by shotgun mass spectrometry. This enables the isolation and identification of a protein target in its native state and its interactome from cells or tissue lysates under physiological conditions. In this chapter, we describe a co-IP protocol for interactome studies that uses an antibody against a protein of interest bound to protein A/G plus agarose beads to isolate a protein complex...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27896668/target-specific-delivery-of-sirna-into-hepatoma-cells-cytoplasm-by-bifunctional-carrier-peptide
#8
Xiaoxuan Liu, Lin Zhu, Jingjing Ma, Xinxiao Qiao, Dunwan Zhu, Lanxia Liu, Xigang Leng
RNA interference (RNAi) is among the most potential approach for the therapy of hepatocellular carcinoma and the major barrier hindering siRNA therapeutics is the low efficiency of delivery to the desired cells. The current study aimed at developing a novel peptide for more efficient hepatoma targeted siRNA delivery, by combining luteinizing hormone-releasing hormone with hepatoma targeting specificity and MPG(△NLS) with cytoplasm-delivery tendency. The developed bifunctional peptide LHRH-MPG(△NLS) and siRNA were mixed together and resulted in LHRH-MPG(△NLS)/siRNA polyplexes through self-assembly...
November 28, 2016: Drug Delivery and Translational Research
https://www.readbyqxmd.com/read/27896233/serum-fructosamine-serum-glycated-albumin-and-serum-glycated-%C3%AE-lipoprotein-in-type-2-diabetes-mellitus-patients-with-and-without-microvascular-complications
#9
Tejaskumar R Kalaria, Habibunnisha B Sirajwala, Mukesh G Gohel
BACKGROUND: Glycation of serum proteins has been proposed as an important mechanism of complications of diabetes but whether there are differences in glycation of different serum proteins and whether it has any correlation with development of microvascular complications has not been studied in depth. This study aimed to assess level of serum fructosamine, glycated albumin and glycated β-lipoprotein in type 2 diabetes mellitus patients with and without microvascular complications and to find out their correlation with diabetes complications...
2016: Journal of Diabetes and Metabolic Disorders
https://www.readbyqxmd.com/read/27895790/corosolic-acid-inhibits-the-proliferation-of-osteosarcoma-cells-by-inducing-apoptosis
#10
Yong Jia, Hua Yuan, Shouqin Shan, Gang Xu, Jie Yu, Chenguang Zhao, Xiang Mou
Corosolic acid (CRA), a pentacyclic triterpene isolated from medicinal herbs, has been reported to exhibit anticancer properties in several cancers. However, the anticancer activity of CRA in osteosarcoma cells is still unclear. In the present study, the inhibitory effect of CRA in osteosarcoma MG-63 cells was investigated, and the results revealed that CRA significantly inhibited the viability of MG-63 cells in a dose- and time-dependent manner. A typical apoptotic hallmark such as DNA ladder was detected by agarose gel electrophoresis following treatment with CRA...
November 2016: Oncology Letters
https://www.readbyqxmd.com/read/27885727/growth-inhibitory-and-proapoptotic-effects-of-l-asparaginase-from-fusarium-culmorum-asp-87-on-human-leukemia-cells-jurkat
#11
Anil Kumar Meghavarnam, Maryam Salah, Meenakshisundaram Sreepriya, Savitha Janakiraman
The objective of this study was to evaluate the anticancer properties of L-asparaginase purified from fungal isolate Fusarium culmorum ASP-87 against human T-cell leukemia cell line (Jurkat). The growth inhibitory and proapoptotic effects of purified L-asparaginase on Jurkat cell lines were investigated by determining its influence on cell viability, colony formation, DNA fragmentation and cell cycle progression. The results revealed that purified L- asparaginase showed significant decrease in cell survival with IC50 value of 90μg/ml(9 IU/ml)...
November 25, 2016: Fundamental & Clinical Pharmacology
https://www.readbyqxmd.com/read/27884430/strengthening-of-c2c12-mouse-myoblasts-against-compression-damage-by-mild-cyclic-compressive-stimulation
#12
Yifei Yao, Arthur Ft Mak
Deep tissue injury (DTI) is a severe kind of pressure ulcers formed by sustained deformation of muscle tissues over bony prominences. As a major clinical issue, DTI affects people with physical disabilities, and is obviously related to the load-bearing capacity of muscle cells in various in-vivo conditions. It is important to provide a preventive approach to help muscle cells from being damaged by compressive stress. In this study, we hypothesized that cyclic compressive stimulation could strengthen muscle cells against compressive damage and enhance the cell plasma membrane resealing capability...
November 16, 2016: Journal of Biomechanics
https://www.readbyqxmd.com/read/27882482/immobilization-and-stabilization-of-beta-xylosidases-from-penicillium-janczewskii
#13
César Rafael Fanchini Terrasan, Maria Romero-Fernández, Alejandro H Orrego, Sandro Martins Oliveira, Benevides Costa Pessela, Eleonora Cano Carmona, José Manuel Guisan
β-Xylosidases are critical for complete degradation of xylan, the second main constituent of plant cell walls. A minor β-xylosidase (BXYL II) from Penicillium janczewskii was purified by ammonium sulfate precipitation (30% saturation) followed by DEAE-Sephadex chromatography in pH 6.5 and elution with KCl. The enzyme presented molecular weight (MW) of 301 kDa estimated by size exclusion chromatography. Optimal activity was observed in pH 3.0 and 70-75 °C, with higher stability in pH 3.0-4.5 and half-lives of 11, 5, and 2 min at 65, 70, and 75 °C, respectively...
November 23, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27882242/development-of-a-loop-mediated-isothermal-amplification-for-diagnosis-of-ascaris-lumbricoides-in-fecal-samples
#14
Esther A Shiraho, Agola L Eric, Ibrahim N Mwangi, Geoffrey M Maina, Joseph M Kinuthia, Martin W Mutuku, Robert M Mugambi, Jackson M Mwandi, Gerald M Mkoji
Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers...
2016: Journal of Parasitology Research
https://www.readbyqxmd.com/read/27876078/simultaneous-detection-and-differentiation-of-three-genotypes-of-brassica-yellows-virus-by-multiplex-reverse-transcription-polymerase-chain-reaction
#15
Xiaoyan Zhang, Yanmei Peng, Ying Wang, Zongying Zhang, Dawei Li, Jialin Yu, Chenggui Han
BACKGROUND: Brassica yellows virus (BrYV), proposed to be a new polerovirus species, three distinct genotypes (BrYV-A, BrYV-B and BrYV-C) have been described. This study was to develop a simple, rapid, sensitive, cost-effective method for simultaneous detection and differentiation of three genotypes of BrYV. RESULTS: In this study, a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneous detection and differentiation of the three genotypes of BrYV...
November 22, 2016: Virology Journal
https://www.readbyqxmd.com/read/27872702/gamma-reactivation-using-the-spongy-effect-of-klf1-binding-site-sequence-an-approach-in-gene-therapy-for-beta-thalassemia
#16
Nasrin Heydari, Laleh Shariati, Hossein Khanahmad, Zahra Hejazi, Mansoureh Shahbazi, Mansoor Salehi
OBJECTIVES: β-thalassemia is one of the most common genetic disorders in the world. As one of the promising treatment strategies, fetal hemoglobin (Hb F) can be induced. The present study was an attempt to reactivate the γ-globin gene by introducing a gene construct containing KLF1 binding sites to the K562 cell line. MATERIALS AND METHODS: A plasmid containing a 192 bp sequence with two repeats of KLF1 binding sites on β-globin and BCL11A promoters was constructed and used to transfect the K562 cell line...
October 2016: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/27871382/relevance-of-substrates-and-products-on-the-desorption-of-lipases-physically-adsorbed-on-hydrophobic-supports
#17
Jose J Virgen-Ortíz, Veymar G Tacias-Pascacio, Daniela B Hirata, Beatriz Torrestiana-Sanchez, Arnulfo Rosales-Quintero, Roberto Fernandez-Lafuente
Lipase B from Candida antarctica (CALB) has been physically immobilized on octyl-agarose via interfacial activation. The incubation of the enzyme in 80% ethanol at pH 5 and 25°C has not significant effect on enzyme activity. Moreover, the hydrolysis of 100mM tributyrin catalyzed by this biocatalyst exhibited a quite linear reaction course. However, a new cycle of tributyrin hydrolysis showed a drastic drop in the activity. SDS-PAGE gels of the supernatant and the biocatalyst showed a significant enzyme desorption after the reaction...
January 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/27865388/quick-and-cheap-miru-vntr-typing-of-mycobacterium-tuberculosis-species-complex-using-duplex-pcr
#18
Memona Yasmin, Stéphanie Le Moullec, Rubina Tabassum Siddiqui, Jessica De Beer, Christophe Sola, Guislaine Refrégier
While minisatellites are usually typed using capillary sequencers or qiaplex systems in developed countries, many low-resource regions cannot afford it. We propose an optimized agarose gel electrophoresis method to genotype Mycobacterium tuberculosis species complex minisatellites in their standardized format (24 MIRU-VNTR). It is based on duplex PCRs combining VNTR loci harboring distinct amplicon sizes whatever the repetition number of each locus. This method performs well both on DNA extracts of good quality and on thermolysates while reducing workload and reagents costs...
December 2016: Tuberculosis
https://www.readbyqxmd.com/read/27854308/characterization-of-chemically-induced-bacterial-ghosts-bgs-using-sodium-hydroxide-induced-vibrio-parahaemolyticus-ghosts-vpgs
#19
Hyun Jung Park, Sung Oh, Nagarajan Vinod, Seongmi Ji, Han Byul Noh, Jung Mo Koo, Su Hyeong Lee, Sei Chang Kim, Ki-Sung Lee, Chang Won Choi
Acellular bacterial ghosts (BGs) are empty non-living bacterial cell envelopes, commonly generated by controlled expression of the cloned lysis gene E of bacteriophage PhiX174. In this study, Vibrio parahaemolyticus ghosts (VPGs) were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH), acetic acid, boric acid, citric acid, maleic acid, hydrochloric acid, and sulfuric acid. The MIC values of the respective chemicals were 3.125, 6...
November 15, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27852838/methylation-specific-polymerase-chain-reaction-pcr-for-gene-specific-dna-methylation-detection
#20
Paul M Lizardi, Qin Yan, Narendra Wajapeyee
Methylation-specific polymerase chain reaction (MS-PCR) is a more rapid way to detect changes in DNA methylation than is bisulfite sequencing. In addition, by incorporating some basic automation, samples can be prepared and analyzed in a 96-well plate format. The method can be used either quantitatively (qRT-PCR-based MethyLight) or qualitatively (using agarose gels) to detect changes in DNA methylation; both are described in this protocol.
November 16, 2016: Cold Spring Harbor Protocols
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