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https://www.readbyqxmd.com/read/29030821/retroviral-transduction-of-quiescent-murine-hematopoietic-stem-cells
#1
Chun Shik Park, H Daniel Lacorazza
Hematopoietic stem cells (HSCs) represent an important target cell population in bone marrow transplantation, cell and gene therapy applications, and the development of leukemia models for research. Because the hematopoietic progeny carries the genetic information of HSCs and replenishes the blood and immune system, corrective gene transfer into HSCs provides an ideal therapeutic approach for many monogenic hematological diseases and a useful tool for studies of HSC function and blood formation in normal and malignant hematopoiesis...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29028502/dota-tetrazine-probes-with-modified-linkers-for-tumor-pretargeting
#2
Tilman Läppchen, Raffaella Rossin, Tiemen R van Mourik, Guillaume Gruntz, Freek J M Hoeben, Ron M Versteegen, Henk M Janssen, Johan Lub, Marc S Robillard
INTRODUCTION: Pretargeted radioimmunoimaging and -therapy approaches building on the bioorthogonal inverse-electron-demand Diels-Alder (IEDDA) reaction between strained trans-cyclooctenes (TCO) and electron-deficient tetrazines (Tz) have yielded impressive results in recent years and have proven a vital alternative to biological pretargeting systems. After improvement of the TCO-antibody conjugates, we here report on our evaluation of a new series of radiolabeled Tz-probes. METHODS: Four new Tz-probes were synthesized, radiolabeled with lutetium-177, and characterized in vitro in terms of lipophilicity, reactivity, and stability in PBS and mouse serum...
September 14, 2017: Nuclear Medicine and Biology
https://www.readbyqxmd.com/read/29027657/practical-issues-of-nutrition-during-continuous-renal-replacement-therapy
#3
Dariusz Onichimowski, Radosław Goraj, Rakesh Jalali, Jolanta Grabala, Ewa Mayzner-Zawadzka, Mirosław Czuczwar
Continuous renal replacement therapy (CRRT) in critically ill patients has significant impact on one's ability to provide efficient nutritional therapy. CRRT may help in the prevention of intestinal oedema and the maintenance of the proper function of the gastrointestinal tract by enabling strict control of the fluid balance. It facilitates early introduction of nutrition via the enteral route, as well as allowing for the composition of high-volume feeding mixtures. It is necessary to take into consideration that during CRRT, together with blood purification of toxic substances, nutritive elements are also eliminated to some extent (micro- and macronutrients)...
October 13, 2017: Anaesthesiology Intensive Therapy
https://www.readbyqxmd.com/read/29027450/-cloning-and-expression-of-scavenger-receptor-class-b-bmscrb8-in-silkworm-bombyx-mori
#4
Yuzu Zhao, Kui Zhang, Mei Tang, Man Xu, Chongyang Li, Guangzhao Pan, Li Shen, Hongjuan Cui, Liqun Yang
Scavenger receptor class B is involved in various indispensable physiological processes, like the formation and inhibition of atherosclerosis or other cardiovascular diseases, innate immune defense and the removal of apoptotic cells. Here, we cloned BmSCRB8, a member of scavenger receptor class B in silkworm. We obtained the full-length cDNA sequence of BmSCRB8 by rapid amplification of cDNA ends (RACE), including 2 668 bp. The ORF of BmSCRB8 is 1 704 bp, encoding 567 amino acids. Online software prediction indicated that the molecular weight of BmSCRB8 is 63...
October 25, 2016: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/29025360/best-practice-of-identification-and-proteomic-analysis-of-extracellular-vesicles-in-human-health-and-disease
#5
Barbara W Sódar, Árpád Kovács, Tamás Visnovitz, Éva Pállinger, Károly Vékey, Gabriella Pocsfalvi, Lilla Turiák, Edit I Buzás
Extracellular vesicles are emerging sources of biomarkers for modern preventive and precision medicine. Extracellular vesicles in body fluids offer a unique opportunity for integrative biomarker approaches due to their complex biocargo that includes proteins, lipids, nucleic acids and metabolites. Mass spectrometry-based proteomics data suggest that a significant portion of human proteins are sorted into extracellular vesicles and amenable for biomarker discovery schemes. Areas covered: this review focuses on key aspects of isolation, quality control and subsequent analysis of blood plasma- and conditioned medium-derived extracellular vesicle proteins, and summarizes the current state-of-the-art in the field...
October 13, 2017: Expert Review of Proteomics
https://www.readbyqxmd.com/read/29023078/polymorphism-of-insulin-like-growth-factor-1-gene-igf1-tasi-igf1-snabi-igf1-rsai-and-the-association-with-daily-gain-of-pesisir-cattle-local-breed-from-west-sumatera-indonesia
#6
Yurnalis, Arnim, Dino Eka Putra
BACKGROUND AND OBJECTIVE: The insulin-like growth factor 1 gene (IGF1) is a candidate gene for marker-assisted selection strategies. Two single nucleotide polymorphism of IGF1 gene namely IGF1/SnaBI and IGF1/TasI has been reported to be associated with production traits in several cattle breeds. One single nucleotide polymorphism in exon 4 namely IGF1/Rsa1 has a significant influence (p<0.01) on birth weight, weaning weight and average daily gain of Bali cattle. The objective of this study was to determine the IGF1/SnaBI, IGF1/TasI and IGF1/RsaI polymorphisms and their associations with average daily gain in Pesisir cattle local breed from west Sumatera Indonesia...
2017: Pakistan Journal of Biological Sciences: PJBS
https://www.readbyqxmd.com/read/29023060/polymorphism-of-glutamate-cysteine-ligase-subunit-catalytic-gclc-gene-in-pulmonary-tuberculosis-patients
#7
Ari Yuniastuti, R Susanti, Dewi Mustikaningtyas
BACKGROUND AND OBJECTIVE: The biomarker of oxidative stress in pulmonary tuberculosis patients has not been found. Oxidative stress occurs due to the low level of antioxidants. Single nucleotide polymorphism of glutamate-cysteine ligase subunit catalytic (GCLC) gene namely -129C/T GCLC has been reported to have an association with a risk factor of oxidative stress' susceptibility. The Objective of this study was to determine the GCLC polymorphism in pulmonary tuberculosis (TB) patient...
2017: Pakistan Journal of Biological Sciences: PJBS
https://www.readbyqxmd.com/read/29022248/coxsackievirus-b4-as-a-causative-agent-of-diabetes-mellitus-type-1-is-there-a-role-of-inefficiently-treated-drinking-water-and-sewage-in-virus-spreading
#8
Waled M El-Senousy, Adel Abdel-Moneim, Mahmoud Abdel-Latif, Mohamed H El-Hefnawy, Rehab G Khalil
This study proposed to detect the enterovirus (EV) infection in children with type 1 diabetes mellitus (T1D) and to assess the role of insufficiently treated water and sewage as sources of viral spreading. Three hundred and eighty-two serum specimens of children with T1D, one hundred serum specimens of children who did not suffer from T1D as control, and forty-eight water and sewage samples were screened for EV RNA using nested RT-PCR. The number of genome copies and infectious units of EVs in raw and treated sewage and water samples were investigated using real-time (RT)-PCR and plaque assay, respectively...
October 11, 2017: Food and Environmental Virology
https://www.readbyqxmd.com/read/28994773/improved-method-for-the-establishment-of-an-in-vitro-blood-brain-barrier-model-based-on-porcine-brain-endothelial-cells
#9
Simone S E Nielsen, Piotr Siupka, Ana Georgian, Jane E Preston, Andrea E Tóth, Siti R Yusof, N Joan Abbott, Morten S Nielsen
The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation...
September 24, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28994210/immunomagnetic-beads-based-isolation-of-erythropoietins-from-urine-and-blood-for-sports-anti-doping-control
#10
Philippe Desharnais, Jean-Francois Naud, Christiane Ayotte
According to World Anti-Doping Agency (WADA) technical document for erythropoiesis stimulating agents (ESA) analysis (TD2014EPO), double- blotting of serum/plasma samples is mandatory for all analysis by isoelectric focusing (IEF) and for the confirmation procedures (CP) performed by SDS-PAGE or SAR-PAGE. The goal is to prevent potential cross-reactions of the secondary antibody with remaining proteins in the purified samples. To this end, we have developed an immunopurification method of ESA in serum/plasma samples using a combination of streptavidin-coated immunomagnetic beads and biotinylated anti-EPO polyclonal antibodies...
October 9, 2017: Drug Testing and Analysis
https://www.readbyqxmd.com/read/28987877/the-anthraquinone-emodin-inhibits-the-non-exported-fikk-kinase-from-plasmodium-falciparum
#11
Benjamin C Lin, Darcy R Harris, Lucy M D Kirkman, Astrid M Perez, Yiwen Qian, Janse T Schermerhorn, Min Y Hong, Dennis S Winston, Lingyin Xu, Alexander M Lieber, Matthew Hamilton, Gabriel S Brandt
The FIKK family of kinases is unique to parasites of the Apicomplexan order, which includes all malaria parasites. Plasmodium falciparum, the most virulent form of human malaria, has a family of 19 FIKK kinases, most of which are exported into the host red blood cell during malaria infection. Here, we confirm that FIKK 8 is a non-exported member of the FIKK kinase family. Through expression and purification of the recombinant kinase domain, we establish that emodin is a relatively high-affinity (IC50=2μM) inhibitor of PfFk8...
September 18, 2017: Bioorganic Chemistry
https://www.readbyqxmd.com/read/28982367/revisiting-escherichia-coli-as-microbial-factory-for-enhanced-production-of-human-serum-albumin
#12
Ashima Sharma, Tapan K Chaudhuri
BACKGROUND: Human serum albumin (HSA)-one of the most demanded therapeutic proteins with immense biotechnological applications-is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals...
October 5, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28980403/recent-progress-in-nucleic-acids-isotachophoresis
#13
REVIEW
Vladimíra Datinská, Ivona Voráčová, Ulrich Schlecht, Jan Berka, František Foret
Progress achieved between 2014-2017 in the extraction and sample preparation of nucleic acid by isotachophoresis is reviewed in this paper. The isolation and purification of nucleic acids is very often compromised by a complex matrix such as blood and other bodily fluids, samples from the scene of crime, fossil samples, etc. While most of the common nucleic acids isolation techniques are based on extraction with inherent limitations with regard to quantitative results, isotachophoretic focusing is a quantitative process with a theoretically unlimited concentration factor...
October 5, 2017: Journal of Separation Science
https://www.readbyqxmd.com/read/28974655/isolation-of-high-molecular-weight-dna-from-mammalian-blood-using-proteinase-k-and-phenol
#14
Michael R Green, Joseph Sambrook
This procedure is the method of choice for purification of genomic DNA from mammalian blood when large amounts of DNA are required, for example, for Southern blotting. The usual yield of DNA from buffy coat lymphocytes isolated from 20 mL of normal blood is ∼250 µg.
October 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28967086/intestinal-mucosal-barrier-dysfunction-in-sap-patients-with-mods-ameliorated-by-continuous-blood-purification
#15
Qing Shen, Zhengrong Li, Shanshan Huang, Liman Li, Hua Gan, Xiao-Gang Du
BACKGROUND: Dysfunction of the intestinal mucosal barrier plays an important role in the pathophysiology of severe acute pancreatitis (SAP). Continuous blood purification (CBP) has been shown to improve the prognosis of SAP patients. In order to investigate the effect of CBP on intestinal mucosal barrier dysfunction in SAP patients with MODS, we conducted in vivo and in vitro experiments to explore the underlying mechanisms. METHODS: The markers for the assessment of intestinal mucosal barrier function including serum diamine oxidase (DAO), endotoxin and intestinal epithelial monolayer permeability were detected during CBP therapy...
September 14, 2017: International Journal of Artificial Organs
https://www.readbyqxmd.com/read/28962889/a-new-clerodane-furano-diterpene-glycoside-from-tinospora-cordifolia-triggers-autophagy-and-apoptosis-in-hct-116-colon-cancer-cells
#16
Neha Sharma, Ashok Kumar, P R Sharma, Arem Qayum, Shashank K Singh, Prabhu Dutt, Satya Paul, Vivek Gupta, M K Verma, N K Satti, R Vishwakarma
ETHNOPHARMACOLOGICAL RELEVANCE: Tinospora cordifolia is a miraculous ayurvedic herb used in the treatment of innumerable diseases such as diabetes, gonorrhea, secondary syphilis, anaemia, rheumatoid arthritis, dermatological diseases, cancer, gout, jaundice, asthma, leprosy, in the treatment of bone fractures, liver & intestinal disorders, purifies the blood, gives new life to the whole body; (rejuvenating herb) and many more. Recent studies have revealed the anticancer potential of this plant but not much work has been done on the anticancer chemical constituents actually responsible for its amazing anticancer effects...
September 27, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28960668/inhibitory-effects-of-some-drugs-on-carbonic-anhydrase-enzyme-purified-from-kangal-akkaraman-sheep-in-sivas-turkey
#17
Ümit M Koçyiğit, Sevgi Durna Daştan, Parham Taslimi, Taner Daştan, İlhami Gülçin
In this study, carbonic anhydrase (CA) enzyme was purified and characterized from blood samples of Kangal Akkaraman sheep and inhibitory properties on certain antibiotics were examined. CA purification was composed of preparation of the hemolysate and conducting the Sepharose-4B-tyrosine-sulfanilamide affinity gel chromatography in having specific activity of 11626 EU mg(-1) , yield of 14.40%, and 242.76-fold purification. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed to assess the enzyme purity and a single band was observed...
September 28, 2017: Journal of Biochemical and Molecular Toxicology
https://www.readbyqxmd.com/read/28959352/production-of-erythropoietin-specific-polyclonal-antibodies
#18
Kourosh Maboudi, Seyedeh Marzieh Hosseini, Mina Sepahi, Hashem Yaghoubi, Shahin Hadadian
BACKGROUND: Erythropoietin, as a principal hormone promotes red blood cell production in bone marrow. Varieties of erythropoietin biosimilar are being produced by recombinant DNA technology in cell cultures. The detection or quantifi cation of these molecules are being performed by diff erent methods which some of theme such as Western blot and enzymelinked immunosorbent assay (ELISA) require specifi c antibodies. High cost, inappropriate shipping (cold chain failures), reduced sensitivity and thus poor detection performance are common pitfalls of using commercial kits for performing immunological tests...
March 2017: Iranian journal of biotechnology
https://www.readbyqxmd.com/read/28956530/multiplexed-genetic-engineering-of-human-hematopoietic-stem-and-progenitor-cells-using-crispr-cas9-and-aav6
#19
Rasmus O Bak, Daniel P Dever, Andreas Reinisch, David Cruz Hernandez, Ravindra Majeti, Matthew H Porteus
Precise and efficient manipulation of genes is crucial for understanding the molecular mechanisms that govern human hematopoiesis and for developing novel therapies for diseases of the blood and immune system. Current methods do not enable precise engineering of complex genotypes that can be easily tracked in a mixed population of cells. We describe a method to multiplex homologous recombination (HR) in human hematopoietic stem and progenitor cells and primary human T cells by combining rAAV6 donor delivery and the CRISPR/Cas9 system delivered as ribonucleoproteins (RNPs)...
September 28, 2017: ELife
https://www.readbyqxmd.com/read/28934494/derivatives-of-bst-like-gss-polymerase-with-improved-processivity-and-inhibitor-tolerance
#20
Igor P Oscorbin, Ekaterina A Belousova, Ulyana A Boyarskikh, Aleksandr I Zakabunin, Evgeny A Khrapov, Maksim L Filipenko
At the moment, one of the actual trends in medical diagnostics is a development of methods for practical applications such as point-of-care testing, POCT or research tools, for example, whole genome amplification, WGA. All the techniques are based on using of specific DNA polymerases having strand displacement activity, high synthetic processivity, fidelity and, most significantly, tolerance to contaminants, appearing from analysed biological samples or collected under purification procedures. Here, we have designed a set of fusion enzymes based on catalytic domain of DNA polymerase I from Geobacillus sp...
September 19, 2017: Nucleic Acids Research
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