Analyze Diet

Topic:Immune Response

The immune response in horses involves a complex network of cells, tissues, and molecules that work together to protect the animal from pathogens and other harmful agents. This process includes both innate and adaptive immune mechanisms. Innate immunity provides the first line of defense and involves components such as physical barriers, phagocytic cells, and the complement system. Adaptive immunity, on the other hand, is characterized by the activation of lymphocytes and the production of antibodies, which provide a targeted response to specific antigens. Key components of the equine immune system include T cells, B cells, and various cytokines that facilitate communication between immune cells. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, regulation, and implications of immune responses in equine health.
Equine tetherin blocks retrovirus release and its activity is antagonized by equine infectious anemia virus envelope protein.
Journal of virology    November 13, 2013   Volume 88, Issue 2 1259-1270 doi: 10.1128/JVI.03148-13
Yin X, Hu Z, Gu Q, Wu X, Zheng YH, Wei P, Wang X.Human tetherin is a host restriction factor that inhibits replication of enveloped viruses by blocking viral release. Tetherin has an unusual topology that includes an N-terminal cytoplasmic tail, a single transmembrane domain, an extracellular domain, and a C-terminal glycosylphosphatidylinositol anchor. Tetherin is not well conserved across species, so it inhibits viral replication in a species-specific manner. Thus, studies of tetherin activities from different species provide an important tool for understanding its antiviral mechanism. Here, we report cloning of equine tetherin and charact...
The equine CD1 gene family is the largest and most diverse yet identified.
Immunogenetics    November 7, 2013   Volume 66, Issue 1 33-42 doi: 10.1007/s00251-013-0741-6
Dossa RG, Alperin DC, Hines MT, Hines SA.The CD1 family is a group of non-polymorphic MHC class I-like molecules that present lipid-based antigens to T cells. Previous work in our laboratory demonstrated that cytotoxic T lymphocytes from immune adult horses recognize lipids from the cell wall of an important equine pathogen, Rhodococcus equi. These findings suggest an important role for the equine CD1 antigen presentation system in protective immune responses to microbial pathogens in the horse. In this study, we characterized and mapped the equine CD1 gene cluster. The equine genome was found to contain 13 complete CD1 genes; seven ...
Expression changes and novel interaction partners of talin 1 in effector cells of autoimmune uveitis.
Journal of proteome research    November 6, 2013   Volume 12, Issue 12 5812-5819 doi: 10.1021/pr400837f
Degroote RL, Hauck SM, Treutlein G, Amann B, Fröhlich KJ, Kremmer E, Merl J, Stangassinger M, Ueffing M, Deeg CA.Autoimmune uveitis is characterized by crossing of blood-retinal barrier (BRB) by autoaggressive immune cells. Equine recurrent uveitis (ERU) is a valuable spontaneous model for autoimmune uveitis and analyses of differentially expressed proteins in ERU unraveled changed protein clusters in target tissues and immune system. Healthy eyes are devoid of leukocytes. In ERU, however, leukocytes enter the inner eye and subsequently destroy it. Molecular mechanisms enabling cell migration through BRB still remain elusive. Previously, we detected decreased talin 1 expression in blood-derived granulocy...
Study of lysozyme resistance in Rhodococcus equi.
Current microbiology    October 30, 2013   Volume 68, Issue 3 352-357 doi: 10.1007/s00284-013-0483-6
Hébert L, Bidaud P, Goux D, Benachour A, Laugier C, Petry S.Lysozyme is an important and widespread component of the innate immune response that constitutes the first line of defense against bacterial pathogens. The bactericidal effect of this enzyme relies on its capacity to hydrolyze the bacterial cell wall and also on a nonenzymatic mechanism involving its cationic antimicrobial peptide (CAMP) properties, which leads to membrane permeabilization. In this paper, we report our findings on the lysozyme resistance ability of Rhodococcus equi, a pulmonary pathogen of young foals and, more recently, of immunocompromised patients, whose pathogenic capacity...
Stimulation of airway neutrophils following dexamethasone administration and equid herpesvirus-2 challenge in horses.
Veterinary journal (London, England : 1997)    October 26, 2013   Volume 199, Issue 1 181-183 doi: 10.1016/j.tvjl.2013.10.018
Richard EA, Pitel PH, Lemaitre L, Jas D, Lekeux P, Pronost S, Fortier G.The aim of this study was to investigate neutrophil stimulation following experimentally-induced airway inflammation in healthy horses. Six horses received dexamethasone and four were then inoculated with equid herpesvirus-2 (EHV-2). Significant neutrophilia was detected in tracheal wash and bronchoalveolar lavage fluid for up to 6 days. Concentrations of neutrophil elastase (NE) and myeloperoxidase (MPO) were significantly increased compared to baseline for up to 14 days in tracheal washes and both markers were significantly correlated with neutrophil counts. Serum levels of surfactant protei...
Equine hyperimmune serum protects mice against Clostridium difficile spore challenge.
Journal of veterinary science    October 18, 2013   Volume 15, Issue 2 249-258 doi: 10.4142/jvs.2014.15.2.249
Yan W, Shin KS, Wang SJ, Xiang H, Divers T, McDonough S, Bowman J, Rowlands A, Akey B, Mohamed H, Chang YF.Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized...
Systemic and mucosal IgE antibody responses of horses to infection with Anoplocephala perfoliata.
Veterinary parasitology    October 16, 2013   Volume 199, Issue 1-2 32-41 doi: 10.1016/j.vetpar.2013.10.005
Pittaway CE, Lawson AL, Coles GC, Wilson AD.Infection of horses with Anoplocephala perfoliata induces a severe inflammatory reaction of the caecal mucosa around the site of parasite attachment adjacent to the ileocecal valve. Lesions show epithelial erosion or ulceration of the mucosa with infiltration by eosinophils, lymphocytes and mast cells leading to oedema, gross thickening and fibrosis of the caecal wall. Despite this evidence of an inflammatory reaction to A. perfoliata within the mucosa of the caecum there is little information about the nature of the local immune response to A. perfoliata. An ELISA which assays serum IgG(T) an...
Influenza: are we protecting our horses effectively?
Equine veterinary journal    October 15, 2013   Volume 45, Issue 6 766-767 doi: 10.1111/evj.12172
Marr CM, Sellon D, Mountford D.No abstract available
Real time RT-PCR analysis of inflammatory mediator expression in recurrent airway obstruction-affected horses.
Veterinary immunology and immunopathology    October 8, 2013   Volume 156, Issue 3-4 190-199 doi: 10.1016/j.vetimm.2013.09.020
Padoan E, Ferraresso S, Pegolo S, Castagnaro M, Barnini C, Bargelloni L.The goal of the present study was to investigate mRNA expression levels of several cytokines and inflammatory mediators in broncho-alveolar lavage (BAL) fluid and respiratory epithelium in recurrent airway obstruction (RAO)-affected horses. RAO, also called heaves, is a common, performance-limiting, equine respiratory disease with clinical signs and pathophysiological similarities to human asthma, and characterized by bronchospasm, neutrophilic infiltration and increased mucus in the airways. Six RAO-affected horses were examined twice within 15 days and seven clinically healthy horses were ex...
Identification of autoantigens in body fluids by combining pull-downs and organic precipitations of intact immune complexes with quantitative label-free mass spectrometry.
Journal of proteome research    October 8, 2013   Volume 12, Issue 12 5656-5665 doi: 10.1021/pr4005986
Merl J, Deeg CA, Swadzba ME, Ueffing M, Hauck SM.Most autoimmune diseases are multifactorial diseases and are caused by the immunological reaction against a number of autoantigens. Key for understanding autoimmune pathologies is the knowledge of the targeted autoantigens, both initially and during disease progression. We present an approach for autoantigen identification based on isolation of intact autoantibody-antigen complexes from body fluids. After organic precipitation of high molecular weight proteins and free immunoglobulins, released autoantigens were identified by quantitative label-free liquid chromatography mass spectrometry. We ...
Identification of potential platelet alloantigens in the Equidae family by comparison of gene sequences encoding major platelet membrane glycoproteins.
Veterinary clinical pathology    October 4, 2013   Volume 42, Issue 4 437-442 doi: 10.1111/vcp.12084
Boudreaux MK, Humphries DM.Platelet alloantigens in horses may play an important role in the development of neonatal alloimmune thrombocytopenia (NAIT). Objective: The objective of this study was to evaluate genes encoding major platelet glycoproteins within the Equidae family in an effort to identify potential alloantigens. Methods: DNA was isolated from blood samples obtained from Equidae family members, including a Holsteiner-Oldenburg cross, a Quarter horse, a donkey, and a Plains zebra (Equus burchelli). Gene sequences encoding equine platelet membrane glycoproteins IIb, IIIa (integrin subunits αIIb and β3), Ia (...
Effect of large colon ischemia and reperfusion on concentrations of calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses.
American journal of veterinary research    September 27, 2013   Volume 74, Issue 10 1281-1290 doi: 10.2460/ajvr.74.10.1281
Grosche A, Morton AJ, Graham AS, Polyak MM, Freeman DE.To determine the effect of large colon ischemia and reperfusion on concentrations of the inflammatory neutrophilic protein calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses. Methods: 6 healthy horses. Methods: Horses were anesthetized, and ischemia was induced for 1 hour followed by 4 hours of reperfusion in a segment of the pelvic flexure of the large colon. Blood samples were obtained before anesthesia, before induction of ischemia, 1 hour after the start of ischemia, and 1, 2, and 4 hours after the start of reperfusion from jugular veins and ve...
Sweat hypersensitivity-induced urticaria and sebaceous adenitis in an American Saddlebred.
Journal of veterinary internal medicine    September 20, 2013   Volume 27, Issue 6 1627-1632 doi: 10.1111/jvim.12198
Lorch G, Calderwood Mays MB, Roberts HA, Isler KK.No abstract available
Plasma D-dimer concentrations during experimental EHV-1 infection of horses.
Journal of veterinary internal medicine    September 20, 2013   Volume 27, Issue 6 1535-1542 doi: 10.1111/jvim.12203
Goehring LS, Soboll Hussey G, Gomez Diez M, Benedict K, Maxwell LK, Morley PS, Sloet van Oldruitenborgh-Oosterbaan MM, Lunn DP.Central nervous system blood vessel thrombosis is a part of the pathogenesis of equid herpesvirus-associated myeloencephalopathy (EHM). D-dimers (DD) are stable breakdown products of cross-linked fibrin, and increased DD-plasma concentrations could reflect the degree of systemic coagulation during EHV-1 infection. Objective: We hypothesized that blood DD concentrations will be increased during periods of EHV-1 fever and viremia, reflecting an activated coagulation cascade with fibrinolysis. Methods: Twenty-eight equids were infected with EHV-1 in 3 experimental infection studies. Three (uninfe...
Investigation of the immune response to autologous, allogeneic, and xenogeneic mesenchymal stem cells after intra-articular injection in horses.
Veterinary immunology and immunopathology    September 18, 2013   Volume 156, Issue 1-2 99-106 doi: 10.1016/j.vetimm.2013.09.003
Pigott JH, Ishihara A, Wellman ML, Russell DS, Bertone AL.Mesenchymal stem cells have demonstrated immunomodulatory capabilities as well as modest efficacy in animal models of joint injury, warranting further study as a potential treatment of joint disease. The goal of the study was to investigate the blood and synovial immune and histologic response to intra-articular injection of autologous, allogeneic, and xenogeneic bone marrow-derived mesenchymal stem cells (MSC) in horses. The study group consisted of 6 five-year-old Thoroughbred mares that had been injected previously with 15 million, genetically modified autologous, allogeneic, or xenogeneic ...
Natural exposure of horses to mosquito-borne flaviviruses in south-east Queensland, Australia.
International journal of environmental research and public health    September 17, 2013   Volume 10, Issue 9 4432-4443 doi: 10.3390/ijerph10094432
Prow NA, Tan CS, Wang W, Hobson-Peters J, Kidd L, Barton A, Wright J, Hall RA, Bielefeldt-Ohmann H.In 2011 an unprecedented epidemic of equine encephalitis occurred in south-eastern (SE) Australia following heavy rainfall and severe flooding in the preceding 2-4 months. Less than 6% of the documented cases occurred in Queensland, prompting the question of pre-existing immunity in Queensland horses. A small-scale serological survey was conducted on horses residing in one of the severely flood-affected areas of SE-Queensland. Using a flavivirus-specific blocking-ELISA we found that 63% (39/62) of horses older than 3 years were positive for flavivirus antibodies, and of these 18% (7/38) had ne...
Phospholipids in sera of horses with summer eczema: lipid analysis of the autoserum preparation used in therapy.
Equine veterinary journal    September 12, 2013   Volume 46, Issue 3 322-327 doi: 10.1111/evj.12135
Hallamaa RE, Batchu KC, Tallberg T.Equine summer eczema, also known as insect bite hypersensitivity, affects horses recurrently during summer months. The treatment of this allergic pruritus is difficult and therefore there is a need for efficacious treatments. Autoserum therapy, based on the use of autogenous serum that is specifically prepared for oral administration and given when the animal shows clinical signs has been introduced recently. Lipids are thought to be responsible for the effect of this therapy. Objective: The main aim of this study was to analyse the phospholipid content of autogenous serum preparations and to ...
Humoral response and antiviral cytokine expression following vaccination of thoroughbred weanlings–a blinded comparison of commercially available vaccines.
Vaccine    September 8, 2013   Volume 31, Issue 45 5216-5222 doi: 10.1016/j.vaccine.2013.08.083
Gildea S, Quinlivan M, Murphy BA, Cullinane A.Previous studies in experimental ponies using interferon gamma (IFN-γ) as a marker for cell mediated immune (CMI) response demonstrated an increase in IFN-γ gene expression following vaccination with an ISCOM subunit, a canarypox recombinant and more recently, an inactivated whole virus vaccine. The objective of this study was to carry out an independent comparison of both humoral antibody and CMI responses elicited following vaccination with all these vaccine presentation systems. Antibody response of 44 Thoroughbred weanlings was monitored for three weeks following the second dose of prima...
Differential CD4+ T-cell responses of allergic and non-allergic subjects to the immunodominant epitope region of the horse major allergen Equ c 1.
Immunology    September 3, 2013   Volume 141, Issue 1 52-60 doi: 10.1111/imm.12166
Kailaanmäki A, Kinnunen T, Kwok WW, Rytkönen-Nissinen M, Randell J, Virtanen T.The responses of allergen-specific CD4(+) T cells of allergic and healthy individuals are still incompletely understood. Our objective was to investigate the functional and phenotypic properties of CD4(+) T cells of horse-allergic and healthy subjects specific to the immunodominant epitope region of the major horse allergen Equ c 1. Specific T-cell lines (TCLs) and clones were generated from peripheral blood mononuclear cells with Equ c 1(143-160), the peptide containing the immunodominant epitope region of Equ c 1. The frequency, proliferative response, cytokine production and HLA restriction...
Role of tumor necrosis factor-α, interferon-γ and Fas-ligand on in vitro nitric oxide activity in the corpus luteum.
Cytokine    August 12, 2013   Volume 64, Issue 1 18-21 doi: 10.1016/j.cyto.2013.07.015
Galvão AM, Szóstek AZ, Skarzynski DJ, Ferreira-Dias GM.Normal reproductive function involves the expression of inflammatory mediators. Regarding the corpus luteum (CL), cytokines promote the cross-talk between immune, vascular and steroidogenic cells, among others. Moreover, TNF, IFNG and FASL were shown to regulate equine CL establishment and regression. We hypothesized that cytokines action on equine CL may be mediated by nitric oxide (NO), through the regulation of endothelial NO synthase (eNOS) expression. TNF increased eNOS mRNA level and NO metabolite (nitrite) production during CL growth. Cytokines combined action (TNF+IFNG+FASL) promoted e...
Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus.
Indian journal of virology : an official organ of Indian Virological Society    August 8, 2013   Volume 24, Issue 3 349-356 doi: 10.1007/s13337-013-0149-9
Singha H, Goyal SK, Malik P, Khurana SK, Singh RK.Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and "Stamping Out" policy. The current internationally accepted test for diagnosis of EIA is the ...
Effect of hay dust extract and cyathostomin antigen stimulation on cytokine expression by PBMC in horses with recurrent airway obstruction.
Veterinary immunology and immunopathology    August 5, 2013   Volume 155, Issue 4 229-237 doi: 10.1016/j.vetimm.2013.07.005
Lanz S, Gerber V, Marti E, Rettmer H, Klukowska-Rötzler J, Gottstein B, Matthews JB, Pirie S, Hamza E.Equine recurrent airway obstruction (RAO) is an inflammatory, obstructive airway disease induced by exposure of susceptible horses to inhaled organic dust particles. The immunological process underlying RAO is still unclear. Previous studies have shown that RAO is linked to the Interleukin-4 receptor (IL-4R) gene in one Warmblood family (F1), but not in another (F2). It has also been shown that in F1, but not in F2, RAO is associated with resistance against parasites, suggesting that this association may have an immuno-genetic basis. Therefore, we hypothesized that the T helper (h)1/Th2/regula...
In vitro induction of functional allergen-specific CD4+ CD25high Treg cells in horses affected with insect bite hypersensitivity.
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology    July 31, 2013   Volume 43, Issue 8 889-901 doi: 10.1111/cea.12131
Hamza E, Akdis CA, Wagner B, Steinbach F, Marti E.Insect bite hypersensitivity (IBH) is a recurrent allergic dermatitis of horses with similarities to human atopic eczema, caused by bites of insects of the genus Culicoides. Previous studies suggested a dysregulated T cell tolerance to Culicoides allergen in IBH-affected horses. Objective: We have investigated whether the suppressive function of CD4(+) CD25(high) cells is impaired in IBH-affected horses and possible ways to restore it. Methods: CD4(+) CD25(-) cells sorted from peripheral blood mononuclear cells (PBMC) were stimulated with irradiated autologous PBMC pulsed with Culicoides or te...
In vitro evaluation of defined oligosaccharide fractions in an equine model of inflammation.
BMC veterinary research    July 22, 2013   Volume 9 147 doi: 10.1186/1746-6148-9-147
Vendrig JC, Coffeng LE, Fink-Gremmels J.Dietary supplementation with oligosaccharides has been proven to be beneficial for health in several mammalian species. Next to prebiotic effects resulting in a modulation of gut micro biota, immunomodulatory effects of oligosaccharides have been documented in vivo. Supplementation with defined oligosaccharide fractions has been shown to attenuate allergic responses and enhance defensive immune responses. Despite the accumulating evidence for immunomodulatory effects, very limited information is available regarding the direct mechanism of action of oligosaccharides. This study aims to elucidat...
Identification of novel immunoreactive proteins of Streptococcus zooepidemicus with potential as vaccine components.
Vaccine    July 8, 2013   Volume 31, Issue 38 4129-4135 doi: 10.1016/j.vaccine.2013.06.100
Velineni S, Timoney JF.Streptococcus zooepidemicus is an important opportunistic pathogen of the equine respiratory and reproductive tracts. A normal tonsillar and mucosal commensal, it becomes invasive under conditions of stress such as virus infection, weaning, high temperature, prolonged transportation and failure of uterine involution. The aim of this study was to evaluate the vaccine potential of several surface exposed and secreted proteins of a novel mucoid clone of SzNC78 (ST-307) from an epizootic of equine respiratory disease. Methods: An expression gene library of SzNC78 was probed with a pool of convales...
Equine herpesviruses type 1 (EHV-1) and 4 (EHV-4)–masters of co-evolution and a constant threat to equids and beyond.
Veterinary microbiology    July 6, 2013   Volume 167, Issue 1-2 123-134 doi: 10.1016/j.vetmic.2013.06.018
Ma G, Azab W, Osterrieder N.The equine herpesviruses type 1 (EHV-1) and 4 (EHV-4) are ubiquitous pathogens that affect horse populations on all continents. Despite widespread vaccination, EHV-1 and EHV-4 infections remain a permanent risk. While the two viruses share a high degree of genetic and antigenic similarity, they differ significantly in host range and pathogenicity. Compared to EHV-4, which mainly infects horses and causes respiratory disease, EHV-1 has a broader host range and can result in respiratory disease, abortions, neonatal death, and equine herpesvirusmyeloencephalopathy (EHM). Recent studies have eluci...
Rhodococcus equi: the many facets of a pathogenic actinomycete.
Veterinary microbiology    July 5, 2013   Volume 167, Issue 1-2 9-33 doi: 10.1016/j.vetmic.2013.06.016
Vázquez-Boland JA, Giguère S, Hapeshi A, MacArthur I, Anastasi E, Valero-Rello A.Rhodococcus equi is a soil-dwelling pathogenic actinomycete that causes pulmonary and extrapulmonary pyogranulomatous infections in a variety of animal species and people. Young foals are particularly susceptible and develop a life-threatening pneumonic disease that is endemic at many horse-breeding farms worldwide. R. equi is a facultative intracellular parasite of macrophages that replicates within a modified phagocytic vacuole. Its pathogenicity depends on a virulence plasmid that promotes intracellular survival by preventing phagosome-lysosome fusion. Species-specific tropism of R. equi fo...
The molecular basis for recognition of bacterial ligands at equine TLR2, TLR1 and TLR6.
Veterinary research    July 4, 2013   Volume 44, Issue 1 50 doi: 10.1186/1297-9716-44-50
Irvine KL, Hopkins LJ, Gangloff M, Bryant CE.TLR2 recognises bacterial lipopeptides and lipoteichoic acid, and forms heterodimers with TLR1 or TLR6. TLR2 is relatively well characterised in mice and humans, with published crystal structures of human TLR2/1/Pam3CSK4 and murine TLR2/6/Pam2CSK4. Equine TLR4 is activated by a different panel of ligands to human and murine TLR4, but less is known about species differences at TLR2. We therefore cloned equine TLR2, TLR1 and TLR6, which showed over 80% sequence identity with these receptors from other mammals, and performed a structure-function analysis. TLR2/1 and TLR2/6 from both horses and hu...
Effects of intravenous lipopolysaccharide infusion on glucose and insulin dynamics in horses with equine metabolic syndrome.
American journal of veterinary research    June 28, 2013   Volume 74, Issue 7 1020-1029 doi: 10.2460/ajvr.74.7.1020
Tadros EM, Frank N, De Witte FG, Boston RC.To test the hypothesis that glucose and insulin dynamics during endotoxemia differ between healthy horses and horses with equine metabolic syndrome (EMS). Methods: 6 healthy adult mares and 6 horses with EMS. Methods: Each horse randomly received an IV infusion of lipopolysaccharide (20 ng/kg [in 60 mL of sterile saline {0.9% NaCl} solution]) or saline solution, followed by the other treatment after a 7-day washout period. Baseline insulin-modified frequently sampled IV glucose tolerance tests were performed 27 hours before and then repeated at 0.5 and 21 hours after infusion. Results were ass...
Effects of equine metabolic syndrome on inflammatory responses of horses to intravenous lipopolysaccharide infusion.
American journal of veterinary research    June 28, 2013   Volume 74, Issue 7 1010-1019 doi: 10.2460/ajvr.74.7.1010
Tadros EM, Frank N, Donnell RL.To test the hypothesis that inflammatory responses to endotoxemia differ between healthy horses and horses with equine metabolic syndrome (EMS). Animals-6 healthy horses and 6 horses with EMS. Methods: Each horse randomly received an IV infusion of lipopolysaccharide (20 ng/kg [in 60 mL of sterile saline {0.9% NaCl} solution]) or saline solution, followed by the other treatment after a 7-day washout period. Baseline data were obtained 30 minutes before each infusion. After infusion, a physical examination was performed hourly for 9 hours and at 15 and 21 hours; a whole blood sample was collect...
1 … 39 40 41 42 43 … 98