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https://www.readbyqxmd.com/read/29617297/the-combinational-use-of-crispr-cas9-and-targeted-toxin-technology-enables-efficient-isolation-of-bi-allelic-knockout-non-human-mammalian-clones
#1
Satoshi Watanabe, Takayuki Sakurai, Shingo Nakamura, Kazuchika Miyoshi, Masahiro Sato
Recent advances in genome editing systems such as clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9) have facilitated genomic modification in mammalian cells. However, most systems employ transient treatment with selective drugs such as puromycin to obtain the desired genome-edited cells, which often allows some untransfected cells to survive and decreases the efficiency of generating genome-edited cells. Here, we developed a novel targeted toxin-based drug-free selection system for the enrichment of genome-edited cells...
April 4, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29580181/decellularised-cartilage-directs-chondrogenic-differentiation-creation-of-a-fracture-callus-mimetic
#2
Wollis Jude Vas, Mittal Shah, Thomas Stephen Blacker, Michael Roland Duchen, Paul Sibbons, Scott John Roberts
Complications that arise from impaired fracture healing have considerable socioeconomic implications. Current research in the field of bone tissue engineering predominantly aims to mimic the mature bone tissue microenvironment. This approach, however, may produce implants that are intrinsically unresponsive to the cues present during the initiation of fracture repair. As such, this study describes the development of decellularised xenogeneic hyaline cartilage matrix in an attempt to mimic the initial reparative phase of fracture repair...
March 27, 2018: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/29492269/mammalian-meat-allergy-following-a-tick-bite-a-case-report
#3
W Landon Jackson
The alpha-gal allergy is an emerging IgE-mediated reaction against the galactose-alpha-1,3-galactose carbohydrate found in mammalian meats. Patients with this condition will develop anaphylactic symptoms 3-6 h after the ingestion of mammalian meat food products such as beef, pork or lamb. The prevalence of this allergy is drastically increasing and severe reactions including anaphylactic shock have been reported, yet many patients experience symptoms for years before a diagnosis is made. We describe the presentation, diagnosis and management of a patient with the alpha-gal allergy in attempt to improve early recognition and management of patients with this condition...
February 2018: Oxford Medical Case Reports
https://www.readbyqxmd.com/read/29486531/establishment-and-identification-of-cell-lines-from-type-o-blood-korean-native-pigs-and-their-efficiency-in-supporting-embryonic-development-via-somatic-cell-nuclear-transfer
#4
Anukul Taweechaipaisankul, Geon A Kim, Jun-Xue Jin, Su Cheong Yeom, Byeong Chun Lee
Due to their close similarities with humans in anatomy, physiology and genetics and handling advantages, miniature pigs are becoming a very attractive model for biomedical research. The purpose of this study was to establish and evaluate blood type O cells derived from Korean native pig (KNP), a typical miniature pig breed in Korea. Total 10 cell lines derived from 8 KNP piglets, adult KNP female pig (kidney and ear tissues) were established. To confirm the blood type O, genomic DNA, fucosyltransferase (FUT) expression and immunofluorescence staining were examined...
February 27, 2018: Journal of Veterinary Science
https://www.readbyqxmd.com/read/29478384/in-vitro-detection-system-to-evaluate-the-immunogenic-potential-of-xenografts
#5
Aila Daugs, Nadine Lehmann, Dilem Eroglu, Martina Meinke, Annina Markhoff, Oliver Bloch
Tissue antigenicity represents the main limitation for the use of xenografts in clinical practice. To eliminate xenoantigens and avoid graft rejection in human, decellularization is often used to remove all immunoreactive components from the extracellular matrix (ECM). After decellularization, acellular scaffolds are required to be investigated regarding the presence of antigens, but commonly used detection methods solely focus on known xenoantigens such as alpha Gal (Galα1,3-Galβ1-4GlcNAc-R) or major histocompatibility complex-I (MHC-I) However, there are unknown xenoantigens, which escape the standard methods...
February 24, 2018: Tissue Engineering. Part C, Methods
https://www.readbyqxmd.com/read/29350022/an-unusual-case-of-positive-sige-to-galactose-alpha-1-3-galactose-from-south-italy
#6
C G Uasuf, A Torina, V Ferrantelli, I Brusca
We report the case of a 38-year-old man who was bitten several times during his life by a tick. He didn't report any previous history of anaphylaxis after the ingestion of red meat. The serum specific IgE showed positivity to α-Gal. The proximity of the bits didn't increase the titer of IgE antibodies to alpha-gal. We could hypothesize that the frequency of the exposure to the tick Corresponding author bites and the amount of tick bites during his lifetime induced a sort of tolerance in this patient.
January 2018: European Annals of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/29273488/repeated-amblyomma-testudinarium-tick-bites-are-associated-with-increased-galactose-%C3%AE-1-3-galactose-carbohydrate-ige-antibody-levels-a-retrospective-cohort-study-in-a-single-institution
#7
Hideo Hashizume, Toshiharu Fujiyama, Takatsune Umayahara, Reiko Kageyama, Andrew F Walls, Takahiro Satoh
BACKGROUND: Alpha-gal syndrome is a hypersensitivity reaction to red meat mediated by IgE antibody specific to galactose-α-1,3-galactose carbohydrate (alpha-gal). Amblyomma tick bites are associated with this condition, but the pathophysiology is not understood. OBJECTIVE: To clarify the mechanism of development of alpha-gal syndrome after tick bites. METHODS: We compared alpha-gal antibody levels between patients with and without a history of tick bites and examined histologic stainings of tick bite lesions between patients with and without detectable alpha-gal IgE antibody...
December 19, 2017: Journal of the American Academy of Dermatology
https://www.readbyqxmd.com/read/29235434/ige-antibodies-to-alpha-gal-in-foresters-and-forest-workers-from-la-rioja-north-of-spain
#8
M Venturini, T Lobera, A Sebastián, A Portillo, J A Oteo
BACKGROUND AND OBJECTIVE: To investigate the prevalence and associated factors to the presence of alpha-gal-specific IgE in a risk group of foresters and forest workers from La Rioja and in a control group. METHODS: A total of 169 workers and 100 individuals who did not recall tick-bites were selected. A questionnaire including demographic data and number of tick bites per year was administered by a physician. Alpha-gal sIgE was assessed with serum samples that had been taken in 2010 using ImmunoCAP®...
December 13, 2017: Journal of Investigational Allergology & Clinical Immunology
https://www.readbyqxmd.com/read/29161766/identification-of-alpha-gal-sensitivity-in-patients-with-a-diagnosis-of-idiopathic-anaphylaxis
#9
M C Carter, K N Ruiz-Esteves, L Workman, P Lieberman, T A E Platts-Mills, D D Metcalfe
IgE antibodies (Ab) specific to galactose-α-1,3-galactose (alpha-gal) are responsible for a delayed form of anaphylaxis that occurs 3-6 hours after red meat ingestion. In a unique prospective study of seventy participants referred with a diagnosis of idiopathic anaphylaxis (IA), six (9%) were found to have IgE to alpha-gal. Upon institution of a diet free of red meat, all patients had no further episodes of anaphylaxis. Two of these individuals had indolent systemic mastocytosis (ISM). Those with ISM had more severe clinical reactions but lower specific IgE to alpha-gal and higher serum tryptase levels, reflective of the mast cell burden...
November 21, 2017: Allergy
https://www.readbyqxmd.com/read/29074067/a-tick-acquired-red-meat-allergy-a-case-series
#10
Joe Kevin Khoury, Neil Christian Khoury, Deborah Schaefer, Anup Chitnis, Getaw Worku Hassen
Allergic reaction is a common clinical picture in the Emergency Department (ED). Most allergic reactions are from food or drugs. A detailed history is an integral aspect of determining the causative agent of an allergy. Galactose-alpha-1,3-galactose (alpha-gal) allergy is a tick-acquired red meat allergy that causes delayed-onset allergic reaction or anaphylaxis due to molecular mimicry. Alpha-gal allergy may not be widely known as a cause of allergic reactions. Lack of universal awareness of this phenomenon in the ED and Urgent Care setting could lead to misdiagnosis, or delayed diagnosis...
October 16, 2017: American Journal of Emergency Medicine
https://www.readbyqxmd.com/read/29057501/different-approaches-to-heart-valve-decellularization-a-comprehensive-overview-of-the-past-30%C3%A2-years
#11
REVIEW
Filippo Naso, Alessandro Gandaglia
Xenogeneic decellularized heart valve scaffolds have the potential to overcome the limitations of existing bioprosthetic heart valves that have limited duration due to calcification and tissue degeneration phenomena. This article presents a review of 30 years of decellularization approaches adopted in cardiovascular tissue engineering, with a focus on the use, either individually or in combination, of different detergents. The safety and efficacy of cell-removal procedures are specifically reported and discussed, as well as the structure and biomechanics of the treated extracellular matrix (ECM)...
October 22, 2017: Xenotransplantation
https://www.readbyqxmd.com/read/29053434/alpha-gal-inactivated-heart-valve-bioprostheses-exhibit-an-anti-calcification-propensity-similar-to-knockout-tissues
#12
Filippo Naso, Ugo Stefanelli, Edward Buratto, Giovanna Lazzari, Andrea Perota, Cesare Galli, Alessandro Gandaglia
BACKGROUND: Glutaraldehyde (GLA) has been used to crosslink bioprosthetic heart valve (BHVs) tissues to enhance their stability, besides ensuring a satisfactory degree of immunological tolerance. Unfortunately, GLA fixation does not guarantee a complete tissue biocompatibility of BHVs in currently used devices. The interaction between preformed human anti-alpha-Gal antibody and alpha-Gal antigens promotes the calcification of GLA-treated alpha-Gal-positive tissue. Recently, an alarming correlation between the presence of the alpha-Gal epitope and a premature BHVs degeneracy was reported...
October 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28972138/-photorhabdus-luminescens-lectin-a-plla-a-new-probe-for-detecting-%C3%AE-galactoside-terminating-glycoconjugates
#13
Ghamdan Beshr, Asfandyar Sikandar, Eva-Maria Jemiller, Nikolai Klymiuk, Dirk Hauck, Stefanie Wagner, Eckhard Wolf, Jesko Koehnke, Alexander Titz
Lectins play important roles in infections by pathogenic bacteria, for example, in host colonization, persistence, and biofilm formation. The Gram-negative entomopathogenic bacterium Photorhabdus luminescens symbiotically lives in insect-infecting Heterorhabditis nematodes and kills the insect host upon invasion by the nematode. The P. luminescens genome harbors the gene plu2096 , coding for a novel lectin that we named PllA. We analyzed the binding properties of purified PllA with a glycan array and a binding assay in solution...
December 1, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28921428/production-of-recombinant-factor-viii-in-human-cell-lines
#14
Daianne Maciely Carvalho Fantacini, Virgínia Picanço-Castro
Human cell lines can produce recombinant proteins much more similar to their natural counterpart, compared to other mammalian cell lines, reducing potential immunogenic reactions. Recombinant proteins produced in nonhuman cells can have in its structure glycan epitopes, such as Galα1,3-Gal (alpha-Gal) and N-glycolylneuraminic acid (Neu5Gc) residues, that are antigenic to humans and can potentially affect the efficacy of the recombinant product. Therefore, the production of recombinant factor VIII (rFVIII) in human cell lines is a new approach to avoid nonhuman glycosylation...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28815010/development-and-characterisation-of-a-low-concentration-sodium-dodecyl-sulphate-decellularised-porcine-dermis
#15
Jack A Helliwell, Daniel S Thomas, Vaia Papathanasiou, Shervanthi Homer-Vanniasinkam, Amisha Desai, Louise M Jennings, Paul Rooney, John N Kearney, Eileen Ingham
The aim of this study was to adapt a proprietary decellularisation process for human dermis for use with porcine skin. Porcine skin was subject to: sodium chloride (1 M) to detach the epidermis, trypsin paste to remove hair follicles, peracetic acid (0.1% v/v) disinfection, washed in hypotonic buffer and 0.1% (w/v) sodium dodecyl sulphate in the presence of proteinase inhibitors followed by nuclease treatment. Cellular porcine skin, decellularised porcine and human dermis were compared using histology, immunohistochemistry, GSL-1 lectin (alpha-gal epitope) staining, biochemical assays, uniaxial tensile and in vitro cytotoxicity tests...
January 2017: Journal of Tissue Engineering
https://www.readbyqxmd.com/read/28752723/red-meat-allergy-induced-by-tick-bites-a-norwegian-case-report
#16
G Arslan Lied
Food allergies, especially delayed hypersensitivity reactions, are often challenging for both patients and clinicians. Here, we report the case of a 64-year-old man who had allergic reactions six hours after eating a meal containing red meat. He reported that he had several tick bites in months before the reaction. High serum specific IgE levels of alpha-gal confirmed the diagnosis of alpha-gal allergic reaction with delayed onset after red meat ingestion caused by tick bite.
July 2017: European Annals of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28670695/meat-allergy-associated-with-galactosyl-%C3%AE-1-3-galactose-%C3%AE-gal-closing-diagnostic-gaps-by-anti-%C3%AE-gal-ige-immune-profiling
#17
U Jappe, S Minge, B Kreft, A Ludwig, B Przybilla, A Walker, R Varga, P Seidel, T Biedermann, W Anemüller, A Kromminga, F Ruëff, H Merk, N Wagner, R Treudler, M Worm, I Waldmann, J Saloga, W M Becker, T Goldmann, T A Platts-Mills, A Homann
BACKGROUND: Glycoproteins and glycolipids of some mammalian species contain the disaccharide galactosyl-α-(1,3)-galactose (α-Gal). It is known that α-Gal is immunogenic in humans and causes glycan-specific IgG and also IgE responses with clinical relevance. α-Gal is part of the IgE-reactive monoclonal therapeutic antibody cetuximab (CTX) and is associated with delayed anaphylaxis to red meat. In this study, different α-Gal-containing analytes are examined in singleplex and multiplex assays to resolve individual sensitization patterns with IgE against α-Gal...
January 2018: Allergy
https://www.readbyqxmd.com/read/28497612/accurate-assessment-of-alpha-gal-syndrome-using-cetuximab-and-bovine-thyroglobulin-specific-ige
#18
Da Woon Sim, Jong Sun Lee, Kyung Hee Park, Kyoung Yong Jeong, Young-Min Ye, Jae-Hyun Lee, Jung-Won Park
SCOPE: IgE against galactose-α-1,3-galactose (α-Gal) causes alpha-gal syndrome. Bovine thyroglobulin (BTG) and cetuximab share this epitope. We aimed to determine the utility of specific IgE (sIgE) against cetuximab as compared to BTG for diagnosing alpha-gal syndrome. METHODS AND RESULTS: Twelve patients with alpha-gal syndrome, 11 patients with immediate beef or pork allergy, 18 asymptomatic individuals with meat sensitization, and 10 non-atopic subjects were enrolled...
May 12, 2017: Molecular Nutrition & Food Research
https://www.readbyqxmd.com/read/28472163/covalent-decoration-of-adenovirus-vector-capsids-with-the-carbohydrate-epitope-%C3%AE-gal-does-not-improve-vector-immunogenicity-but-allows-to-study-the-in-vivo-fate-of-adenovirus-immunocomplexes
#19
Ramona F Kratzer, Sigrid Espenlaub, Andrea Hoffmeister, Matthias W Kron, Florian Kreppel
Adenovirus-based vectors are promising tools for genetic vaccination. However, several obstacles have to be overcome prior to a routine clinical application of adenovirus-based vectors as efficacious vectored vaccines. The linear trisaccharide epitope αGal (alpha-Gal) with the carbohydrate sequence galactose-α-1,3-galactosyl-β-1,4-N-acetylglucosamine has been described as a potent adjuvant for recombinant or attenuated vaccines. Humans and α-1,3-galactosyltransferase knockout mice do not express this epitope...
2017: PloS One
https://www.readbyqxmd.com/read/28447914/diagnosis-of-life-threatening-alpha-gal-food-allergy-appears-to-be-patient-driven
#20
Mary Grace Flaherty, Samantha Jan Kaplan, Maya R Jerath
OBJECTIVE: Patients exhibiting life-threatening symptoms associated with the alpha-gal food allergy (delayed urticaria or anaphylaxis due to mammalian meat) are frequently undiagnosed, causing unnecessary emergency department (ED) and health care visits, and extensive pain and suffering. This study aimed to determine the path to diagnosis experienced by alpha-gal patients. METHODS: Semistructured interviews were conducted from March to June 2016 with a chronological systematic sample of approximately 10% of patients diagnosed with alpha-gal and treated by the University of North Carolina Allergy and Immunology Clinic (n = 28)...
October 2017: Journal of Primary Care & Community Health
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