ABSTRACT
BACKGROUND: Historically, cryopreservation of equine embryos >300 µm gave poor pregnancy rates until researchers collapsed the blastocoele cavity and aspirated the blastocoele fluid prior to vitrification. OBJECTIVE: To determine if aspiration of the blastocoele fluid prior to vitrification is essential for post warming survival. STUDY DESIGN: In vivo experiments. METHODS: Fifty embryos were recovered on day 7-8 and washed in holding medium (HM; M-199HEPES + 20% FBS + antibiotics). Embryos were punctured using a micromanipulator mounted 30 µm biopsy needle; following this 28 had >90% of their blastocoelic fluid actively aspirated while the remaining 22 were not-aspirated. Embryos were then vitrified using a two-step process with increasing concentrations of DMSO and ethylene glycol (7.5-15% v:v), and 0.5 mol/L sucrose in the final solution before being loaded onto a Cryolock device and plunged into liquid nitrogen. The embryos were warmed by plunging the Cryolock tip into HM with 1 mol/L sucrose at 37°C. After 1 min, the embryos were transferred to HM + 0.5 mol/L sucrose at RT for 4 min before transfer into HM for a further 4 min prior to transfer to a recipient mare. RESULTS: Mean (±s.e.) embryo diameter was not significantly different between the punctured and punctured plus aspirated group (646.4 ± 61.7 vs. 754.8 ± 59.1 µm, respectively; P = 0.215). Nonaspirated and aspirated embryos gave pregnancy rates of 10/22 (45%) and 21/28 (75%) respectively (P = 0.061). Sub-dividing embryos on the basis of size showed that vitrification of larger embryos (>550 µm) yielded a significantly higher pregnancy rate when they were aspirated vs. not-aspirated (13/18 [72%] vs. 1/10 [10%], respectively; P = 0.006), whereas there was no difference for smaller embryos (8/10 [80%] vs. 9/12 [75%], respectively; P = 0.8). MAIN LIMITATIONS: Group sizes are limited. CONCLUSION: Aspiration of blastocoele fluid from embryos ≤550 µm is not a pre-requisite for successful vitrification. The Summary is available in Spanish - see Supporting Information.
Subject(s)
Blastocyst/physiology , Cryopreservation/veterinary , Embryo, Mammalian/physiology , Vitrification , Animals , Cryopreservation/methods , Embryonic Development , HorsesABSTRACT
This study was designed to investigate the effect of pterostilbene (PTS) on cardiac oxidative stress in vitro, as this is a simple and promising methodology to study cardiac disease. Cardiac myoblasts (H9c2 cells) and homogenised cardiac tissue were incubated with the PTS and cyclodextrin (PTS + HPßCD) complex for 1 and 24 h, respectively, at concentrations of 50µM for the cells and 25 and 50µM for cardiac tissue. The PTS + HPßCD complex was used to increase the solubility of PTS in water. After the pretreatment period, cardiomyoblasts were challenged with hydrogen peroxide (6.67µM) for 10 min, while cardiac tissue was submitted to a hydroxyl radical generator system (30 min). Cellular viability, oxidative stress biomarkers (e.g. total reactive oxygen species (ROS), carbonyl assay and lipoperoxidation) and the antioxidant response (e.g. sulfhydryl and the antioxidant enzyme activities of superoxide dismutase, catalase and glutathione peroxidase) were evaluated. In cardiomyoblasts, the PTS + HPßCD complex (50µM) increased cellular viability. Moreover, the PTS + HPßCD complex also significantly increased sulfhydryl levels in the cells submitted to an oxidative challenge. In cardiac tissue, lipid peroxidation, carbonyls and ROS levels were significantly increased in the groups submitted to oxidative damage, while the PTS + HPßCD complex significantly reduced ROS levels in these groups. In addition, the PTS + HPßCD complex also provoked increased catalase activity in both experimental protocols. These data suggest that the PTS + HPßCD complex may play a cardioprotective role through a reduction of ROS levels associated with an improved antioxidant response.
Subject(s)
Antioxidants/pharmacology , Heart/drug effects , Homeostasis/drug effects , Myoblasts, Cardiac/drug effects , Stilbenes/pharmacology , Animals , Antioxidants/chemistry , Apoptosis/drug effects , Catalase/metabolism , Cell Survival/drug effects , Cyclodextrins/chemistry , Glutathione Peroxidase/metabolism , Homeostasis/physiology , Hydrogen Peroxide/antagonists & inhibitors , Hydrogen Peroxide/pharmacology , Lipid Peroxidation/drug effects , Male , Myoblasts, Cardiac/cytology , Myoblasts, Cardiac/metabolism , Myocytes, Cardiac/cytology , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , Oxidation-Reduction , Rats , Rats, Wistar , Reactive Oxygen Species/antagonists & inhibitors , Reactive Oxygen Species/metabolism , Stilbenes/chemistry , Superoxide Dismutase/metabolismABSTRACT
In this paper a method to evaluate the presence of microorganisms of the coliform group in water samples using the ultra-weak bioluminescence (UWB) is proposed. A series of UWB measurements and optical density measurements from cultures of both a set of standard E. coli strain samples, and a set of water samples from a river near Curitiba City in Brazil were performed. All samples were previously incubated at 37°C for 11h in nutritive medium before the temporal UWB emission profiles data were acquired for a period of 24h inside a dark chamber of an especially implemented instrumentation capable of doing photon counting measurements. For the optical density measurements, a spectrophotometer was used to acquire the growth kinetics of those cultures for a period of 13h, and the results compared to the UWB profiles. Periodic time-components analysis of the UWB data from both the set of standard E. coli samples and the set of the river's water samples were performed and compared to each other. The results have shown that the UWB temporal profiles resemble in some way the growth kinetics curve and the periodic time-components analysis is an effective way to discriminate between contaminated and non-contaminated samples, therefore the method may be viable for detecting coliforms in water samples in less time than usual methods.
Subject(s)
Environmental Monitoring/methods , Luminescent Measurements , Water Microbiology , Water/chemistry , Escherichia coli/growth & development , Photons , Rivers/microbiologySubject(s)
Hemoglobin E/genetics , Hemoglobin, Sickle/genetics , beta-Globins/genetics , Adult , Asymptomatic Diseases , Base Sequence , Brazil , Child, Preschool , Female , Genetic Testing , Hemoglobin E/analysis , Hemoglobin, Sickle/analysis , Heterozygote , Humans , Infant , Infant, Newborn , Male , Molecular Sequence Data , Neonatal ScreeningABSTRACT
INTRODUCTION: It has been shown that the innate immune system mediates acute lung inflammation triggered by intestinal trauma. Sexual dimorphism modulates the profile of TH1 and TH2 lymphocytes, and accordingly sex hormones may modulate acute lung inflammation by intestinal ischemia/reperfusion (I/R). Studies indicate that female rats are relatively resistant to organ injury caused by hemorrhagic shock and that the gut of female is more resistant than that of the male to deleterious effects of ischemic injury. At the present study, we investigated the effect of estradiol (E(2)) on the lung inflammation after intestinal I/R and its interaction with the nitric oxide (NO) pathway. METHODS: Anesthetized female rats submitted or not to 7 days ovariectomy (OVx) were subjected to occlusion of the superior mesenteric artery during 45 min, followed by 2 h of reperfusion. Groups of rats were treated with E(2) (17ß-estradiol, 280 µg/kg, s.c.) 24 h before ischemia and/or with the nonselective NO synthase inhibitor L-NAME (Nω-nitro-L-arginine methyl ester hydrochloride) (5 mg/kg, i.v.). In a parallel set of experiments, the selective NO synthase inhibitor, aminoguanidine (50 mg/kg i.v.), was given 1 h before ischemia. In all groups, lung vascular permeability (LVP) was assessed using the Evans blue dye extravasation method, neutrophil recruitment to the tissues by the standard myeloperoxidase (MPO) method, and endothelial NO synthase (eNOS) protein expression by Western blot. RESULTS: In OVx rats, LVP and MPO were increased after intestinal I/R as compared with intact controls. Estradiol reverted the LVP, but not MPO. Aminoguanidine reduced LVP in OVx rats. The E(2) protective effect on LVP was abolished by L-NAME; moreover, an increase in LVP even when compared with OVx rats treated only with L-NAME was observed. In addition, lung eNOS protein expression was reduced in OVx-I/R rats in comparison to intact controls and the E(2) inhibited this effect. CONCLUSIONS: Estradiol treatment is able to reduce lung inflammation due to intestinal I/R, but with the concomitant blockade of NOS activity, this effect is abolished. Nitric oxide probably reduces the vascular deleterious effects of intestinal I/R, and E(2) pretreatment reduces lung inflammation after intestinal I/R and exerts these effects by modulating eNOS protein expression in the lungs.
Subject(s)
Estradiol/therapeutic use , Intestines/blood supply , Nitric Oxide/metabolism , Pneumonia/drug therapy , Pneumonia/metabolism , Animals , Female , Male , Rats , Rats, WistarABSTRACT
The objective of this study was to explore the influence of the renin-angiotensin system on cardiac prooxidants and antioxidants levels and its association to autonomic imbalance induced by hyperthyroidism. Male Wistar rats were divided into four groups: control, losartan (10mg/kg/day by gavage, 28 day), thyroxine (T4) (12 mg/L in drinking water for 28 days), and T4+losartan. Spectral analysis (autonomic balance), angiotensin II receptor (AT1R), NADPH oxidase, Nrf2 and heme-oxygenase-1 (HO-1) myocardial protein expression, and hydrogen peroxide (H2O2) concentration were quantified. Autonomic imbalance induced by hyperthyroidism (~770%) was attenuated in the T4+losartan group (~32%) (P<0.05). AT1R, NADPH oxidase, H2O2, as well as concentration, Nrf2 and HO-1 protein expression were elevated (~172%, 43%, 40%, 133%, and 154%, respectively) in T4 group (P<0.05). H2O2 and HO-1 levels were returned to control values in the T4+losartan group (P<0.05). The overall results demonstrate a positive impact of RAS blockade in the autonomic control of heart rate, which was associated with an attenuation of H2O2 levels, as well as with a reduced counter-regulatory response of HO-1 in experimental hyperthyroidism.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Hyperthyroidism/metabolism , Reactive Oxygen Species/metabolism , Receptor, Angiotensin, Type 1/physiology , Angiotensin II Type 1 Receptor Blockers/therapeutic use , Animals , Heart Rate/drug effects , Heart Rate/physiology , Hyperthyroidism/drug therapy , Male , Rats , Rats, WistarABSTRACT
The objective was to compare a leukocyte esterase (LE) test with endometrial cytology (EC) for diagnosis of subclinical endometritis in dairy cows. The relationship between subsequent fertility and the uterine (Ut) and cervical (Cx) leukocyte esterase activity was determined by the odds of pregnancy by 90 days in milk (DIM). Holstein cows (N = 218) without clinical endometritis and between 21 and 47 DIM from five commercial dairy herds were sampled for uterine and cervical leukocyte esterase activity and EC by cytobrush. To test the effect of time, cows were grouped into early (21-31 DIM) and late (32-47 DIM) animals. There was a slight agreement between UtLE and CxLE (weighted κ = 0.37). The percentage of neutrophils was correlated with esterase score either from the uterus (UtLE; P = 0.0001) or cervix (CxLE; P = 0.002). The percentage of neutrophils on EC (P < 0.001), the UtLE score (P < 0.0001), and the CxLE (P = 0.0009) diminished as DIM increased. Neither CxLE nor UtLE were statistically associated with pregnancy at 90 DIM. However, between 32 and 47 DIM, the percentage of neutrophils on EC and odds of pregnancy at 90 DIM were associated (P = 0.04). For the same interval, based on receiver/response operating characteristics analysis, the optimal cutoff was >6.7% neutrophils to classify cows with subclinical endometritis. In conclusion, uterine LE activity was correlated with percentage of neutrophils as determined by EC, but not with odds of pregnancy. Subclinical endometritis (>6.70% neutrophils) diagnosed by EC between 32 and 47 DIM was associated with reduced odds of pregnancy.
Subject(s)
Carboxylic Ester Hydrolases/analysis , Cattle Diseases/diagnosis , Diagnostic Techniques, Obstetrical and Gynecological , Endometritis/diagnosis , Endometrium/pathology , Animals , Asymptomatic Infections , Cattle , Cattle Diseases/etiology , Dairying , Diagnostic Techniques, Obstetrical and Gynecological/veterinary , Endometritis/pathology , Endometritis/veterinary , Endometrium/cytology , Female , Postpartum Period/physiology , Pregnancy , Puerperal Disorders/diagnosis , Puerperal Disorders/etiology , Puerperal Disorders/veterinary , Reagent Kits, Diagnostic , Urinalysis/methods , Urinalysis/veterinaryABSTRACT
Biocompatibilidade é a capacidade de um material exercer funções específicas quando aplicado em contato com tecidos vivos de determinado hospedeiro, sem, contudo, causar danos ou prejuízo ao mesmo. Este trabalho objetivou determinar a biocompatibilidade in vivo e in vitro do extrato hidroalcoólico do Cissus sicyoides L - Vitaceae. Foram utilizados 30 ratos (Rattus novergicus albinus wistar), com idade entre 45 e 90 dias e pesando entre 170 e 260 g. Os animais foram divididos em 3 grupos (A1, A2 e A3) de 6 animais cada para o teste in vivo, os quais foram sacrificados com 2, 4 e 6 dias, respectivamente. Para o teste in vitro, foram utilizados 12 animais para obtenção do índice de aderência e da capacidade fagocítica dos macrófagos de ratos do grupo controle e do grupo experimental. Nos resultados encontrados no teste in vivo, conclui-se que o extrato apresentou-se biocompatível, visto que não provocou alterações significativas no tecido. Já no teste in vitro, o mesmo não se apresentou biocompatível, pois o extrato puro apresentou índice de aderência baixo (7,1) e taxa de fagocitose elevada (35,7), indicando diferença significante quando comparado ao controle. Porém, quando diluído, o extrato se mostrou inócuo, devido ao aumento dos valores do índice de aderência nas concentrações de 1/10 (61,4) e 1/100 (74,3) nos ensaios, as quais não apresentaram diferença significante quando comparadas ao controle. Após a análise dos dados, concluiu-se que a solução diluída do extrato hidroalcoólico do Cissus sicyoides L. não causa danos ou prejuízos. Entretanto, como nem todos os efeitos farmacológicos foram testados no presente trabalho, não se pode inferir automaticamente que ele é biocompatível em todos os casos.
Biocompatibility is the ability of a material to perform specifictasks when applied to living tissues without causing damage or injuries to it. Thus, this study aimed at determining the in vivo and in vitro biocompatibility of Cissus sicyoides L. - Vitaceae hydroalcoholic extracts. A total of 30 rats (Rattus norvegicus Albinus Wistar), with ages ranging from 45 to 90 days and weighing between 170 and 260g were used. The animals were divided into 3 groups (A1, A2 and A3) with 6 animals each, for the in vivo test, which were sacrificed after 2, 4 and 6 days, respectively. For in vitro test, 12 animals were used to obtain the index of adherence and phagocytic ability of macrophages of rats from the control and the experimental groups. In results found for the in vivo test, it was concluded that the extract was biocompatible, whereas no significant changes were observed in the tissue. As to the in vitro test, the extract was not biocompatible, since the pure extract showed a low rate of adherence (7.1) and a high rate of phagocytosis (35.7), indicating a significant difference when compared to the control group. However, when diluted, the extract was shown to be harmless, due to an increase in the values of the adherence index at the following concentrations : 1/10 (61.4) and 1/100 (74.3) in the tests, which showed no significant differences when compared to the control group. After analyzing the data, it was concluded that since the infusion of the plant is a kind of dilution, its use does not cause any harm to the body. A new study is necessary at the moment to possibily demonstrate its effects on the long term.
Subject(s)
Animals , Male , Female , Rats , Materials Testing/instrumentation , Vitaceae/classification , Plants, Medicinal/adverse effects , Biocompatible Materials/analysisABSTRACT
Determinou-se a digestibilidade intestinal (DI) da proteína de vários coprodutos do biodiesel nas formas de farelo e torta. Foram avaliados oito coprodutos: tortas e farelos de pinhão manso, nabo forrageiro, tremoço, algodão. Os coprodutos foram incubados no rúmen por 16 horas, e os resíduos não degradados no rúmen submetidos à digestão enzimática com solução de pepsina e pancreatina para a determinação da DI. Ainda, nos resíduos da incubação ruminal, foram determinadas: degradabilidade da matéria seca (DR), proteína degradável no rúmen (PDR) e proteína não degradável no rúmen (PNDR). A digestibilidade intestinal da proteína para os coprodutos do biodiesel variou de 2,4 a 48,6%. Todos os coprodutos avaliados caracterizaram-se como alimentos de alto teor proteico, sendo considerados de alta PDR, e apresentaram baixa digestibilidade intestinal da proteína. A DI da proteína dos coprodutos do biodiesel na forma de torta foi maior em comparação com a dos farelos. A torta e o farelo de algodão apresentaram os maiores coeficientes de DI.
The objective of this research was to determine intestinal protein digestibility (ID) of some biodiesel by-products in the form of cakes and the meals. Eight by-products were: cakes and meals of physic nut, turnip, lupine, cotton cake, cottonseed meal. The by-products were incubated in the rumen for 16 hours, were the undegradable rumen residues were submitted to enzymatic digestion with pepsin and pancreatin solution for the determination of ID. In the incubation residues the following was also determined: dry matter degradability (RD), rumen degradable protein (RDP) and rumen undegradable protein (RUP). The intestinal protein digestibility of biodiesel by-products ranged from 2.4 to 48.6%. All the by-products evaluated in this study were characterized as high protein sources and were considered high-RDP. The by-products presented low intestinal protein digestibility. The ID protein of biodiesel by-products was higher in the cakes than the meals. The by-products evaluated, the cottonseed cake and meal presented the highest ID coefficients.
Subject(s)
Animals , Digestion , Ruminants , Rumen/metabolism , Proteins/analysis , ProteinsABSTRACT
OBJECTIVES: To evaluate citrated recalcified thromboelastography (TEG) in healthy newborn foals, and to determine intra-assay, inter-individual and intra-individual (at 12 h, 24 h and 7 days after birth) variations. Additionally, to compare TEG variables, haematological values and conventional coagulation profiles from healthy, sick non-septic, and septic foals. DESIGN: Prospective study. METHODS: The study group comprised 18 healthy, 15 sick non-septic and 17 septic foals. Two citrated (3.2%; 1 : 9 anticoagulant : blood ratio) blood samples were submitted for haemostatic evaluation using a TEG analyser and conventional coagulation profile. TEG values (R time (R), K time (K), angle (α), maximum amplitude (MA) and G value (G)), complete blood count (CBC) and conventional coagulation profile (prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen concentration (Fib) and antithrombin (AT)) were evaluated. Signalment, presenting complaint, sepsis scores, blood culture results and outcome were taken from the medical records of the sick foals. RESULTS: Mean values ± SD for TEG variables in healthy neonatal foals were: R = 11.82 ± 5.35 min, K = 3.06 ± 1.34 min, α= 51.19 ± 12.66 degrees, MA = 55.06 ± 6.67 mm and G = 6361 ± 1700 dyn/cm(2) . Mean coefficients of variation for intra-assay/inter-individual/intra-individual in healthy foals were: R = 3.5/45.2/43.1%; K = 5.3/58.7/28.7%; α= 1.5/24.7/11.9%; MA = 0.3/12.1/6.1%; G = 1.6/26.7/14.7%. Septic foals had significantly greater α, MA and G values than sick non-septic foals, and significantly greater MA and G than healthy foals, changes that are consistent with hypercoagulability. Weak correlations were detected between TEG variables and haematological or haemostatic values. CONCLUSIONS: TEG could be used to provide additional information about the haemostatic system in equine neonates.
Subject(s)
Horse Diseases/blood , Horses/blood , Sepsis/veterinary , Thrombelastography/veterinary , Animals , Animals, Newborn , Blood Coagulation , Citrates , Female , Hemostasis , Male , Prospective Studies , Reproducibility of Results , Sensitivity and Specificity , Thrombelastography/methods , Thrombelastography/standards , ThromboplastinABSTRACT
The endothelium plays a vital role in maintaining circulatory homeostasis by the release of relaxing and contracting factors. Any change in this balance may result in a process known as endothelial dysfunction that leads to impaired control of vascular tone and contributes to the pathogenesis of some cardiovascular and endocrine/metabolic diseases. Reduced endothelium-derived nitric oxide (NO) bioavailability and increased production of thromboxane A2, prostaglandin H2 and superoxide anion in conductance and resistance arteries are commonly associated with endothelial dysfunction in hypertensive, diabetic and obese animals, resulting in reduced endothelium-dependent vasodilatation and in increased vasoconstrictor responses. In addition, recent studies have demonstrated the role of enhanced overactivation of ß-adrenergic receptors inducing vascular cytokine production and endothelial NO synthase (eNOS) uncoupling that seem to be the mechanisms underlying endothelial dysfunction in hypertension, heart failure and in endocrine-metabolic disorders. However, some adaptive mechanisms can occur in the initial stages of hypertension, such as increased NO production by eNOS. The present review focuses on the role of NO bioavailability, eNOS uncoupling, cyclooxygenase-derived products and pro-inflammatory factors on the endothelial dysfunction that occurs in hypertension, sympathetic hyperactivity, diabetes mellitus, and obesity. These are cardiovascular and endocrine-metabolic diseases of high incidence and mortality around the world, especially in developing countries and endothelial dysfunction contributes to triggering, maintenance and worsening of these pathological situations.
Subject(s)
Cardiovascular Diseases/physiopathology , Endocrine System Diseases/physiopathology , Endothelium, Vascular/physiopathology , Metabolic Diseases/physiopathology , Nitric Oxide Synthase Type III/metabolism , Animals , Cardiovascular Diseases/metabolism , Diabetes Mellitus/metabolism , Diabetes Mellitus/physiopathology , Endocrine System Diseases/metabolism , Endothelium, Vascular/metabolism , Endothelium-Dependent Relaxing Factors/physiology , Humans , Nitric Oxide/biosynthesis , Obesity/metabolism , Obesity/physiopathology , RatsABSTRACT
The endothelium plays a vital role in maintaining circulatory homeostasis by the release of relaxing and contracting factors. Any change in this balance may result in a process known as endothelial dysfunction that leads to impaired control of vascular tone and contributes to the pathogenesis of some cardiovascular and endocrine/metabolic diseases. Reduced endothelium-derived nitric oxide (NO) bioavailability and increased production of thromboxane A2, prostaglandin H2 and superoxide anion in conductance and resistance arteries are commonly associated with endothelial dysfunction in hypertensive, diabetic and obese animals, resulting in reduced endothelium-dependent vasodilatation and in increased vasoconstrictor responses. In addition, recent studies have demonstrated the role of enhanced overactivation ofβ-adrenergic receptors inducing vascular cytokine production and endothelial NO synthase (eNOS) uncoupling that seem to be the mechanisms underlying endothelial dysfunction in hypertension, heart failure and in endocrine-metabolic disorders. However, some adaptive mechanisms can occur in the initial stages of hypertension, such as increased NO production by eNOS. The present review focuses on the role of NO bioavailability, eNOS uncoupling, cyclooxygenase-derived products and pro-inflammatory factors on the endothelial dysfunction that occurs in hypertension, sympathetic hyperactivity, diabetes mellitus, and obesity. These are cardiovascular and endocrine-metabolic diseases of high incidence and mortality around the world, especially in developing countries and endothelial dysfunction contributes to triggering, maintenance and worsening of these pathological situations.
Subject(s)
Animals , Humans , Rats , Cardiovascular Diseases/physiopathology , Endocrine System Diseases/physiopathology , Endothelium, Vascular/physiopathology , Metabolic Diseases/physiopathology , Nitric Oxide Synthase Type III/metabolism , Cardiovascular Diseases/metabolism , Diabetes Mellitus/metabolism , Diabetes Mellitus/physiopathology , Endocrine System Diseases/metabolism , Endothelium, Vascular/metabolism , Endothelium-Dependent Relaxing Factors/physiology , Nitric Oxide/biosynthesis , Obesity/metabolism , Obesity/physiopathologyABSTRACT
Twenty-one healthy greyhounds with no history or clinical signs of bleeding disorders, and no abnormalities on physical examination, complete blood count, serum biochemistry profiles (in dogs more than five years of age), and SNAP-4DX test for vector borne diseases underwent routine gonadectomies at the Ohio State University Veterinary Teaching Hospital. Blood samples were collected 24 hours before and after surgery by jugular venepuncture for thromboelastography and haemostasis assays (prothrombin time [PT], activated partial thromboplastin time [aPTT], fibrinogen concentration). The magnitude of the bleeding in each patient was estimated using a bleeding scoring system recently validated in greyhounds. Eight dogs were classified as bleeders and 13 as non-bleeders. Thromboelastograph (TEG) tracings in bleeders were different to that of non-bleeders. Neither sex (odds ratio [OR]: 0.148, P=0.05), haematocrit (OR: 0.907, P=0.39), platelet count (OR: 0.996, P=0.65) or age (OR: 0.949, P=0.83) were predictors of the outcome. None of the variables that evaluated clot kinetics, and fibrinolysis (that is, aPTT OR: 0.781, P=0.51; PT OR: 1.337, P=0.63; TEG(R) OR: 1.269, P=0.06; TEG(K) OR: 1.696, P=0.05; TEG(LY60) OR: 1.028, P=0.81) were able to predict the bleeding episodes. Only the TEG variables that represent the fibrin cross-linking of the clot (TEG(angle) OR: 0.903, P=0.03); and the strength of the clot (TEG(MA) OR: 0.833, P=0.03) were considered predictors of the outcome.
Subject(s)
Blood Coagulation Tests/veterinary , Castration/veterinary , Dogs/blood , Hemorrhage/veterinary , Hemostasis/physiology , Thrombelastography/veterinary , Animals , Blood Cell Count/veterinary , Blood Chemical Analysis/veterinary , Dogs/surgery , Female , Hemorrhage/blood , Male , Reference ValuesSubject(s)
Blood Gas Analysis/veterinary , Dogs/blood , Pedigree , Animals , Breeding , Female , Male , SpainABSTRACT
A novel gene coding for a LipA-like lipase with 283 amino acids and a molecular mass of 32 kDa was isolated and characterized from a metagenomic library prepared from mangrove sediment from the south Brazilian coast. LipA was 52% identical to a lipolytic enzyme from an uncultured bacterium and shared only low identities (< or =31%) with lipases/esterases from cultivable microorganisms. Phylogenetic analysis showed that LipA, together with an orthologous protein from an uncultured bacterium, forms a unique branch within family I of true lipases, thereby constituting a new lipase subfamily. Activity determination using crude extracts of Escherichia coli bearing the lipA gene revealed that this new enzyme has a preference for esters with short-chain fatty acids (C < or = 10) and has maximum activity against p-nitrophenyl-caprate (chain length C10, 0.87 U/mg protein). The optimum pH of LipA was 8.0, and the enzyme was active over a temperature range of 20 to 35 degrees C, with optimum activity against p-nitrophenyl-butyrate at 35 degrees C and pH 8.0.
Subject(s)
Gene Library , Geologic Sediments/chemistry , Lipase/isolation & purification , Metagenomics/methods , Rhizophoraceae , Seawater , Brazil , DNA/isolation & purification , Enzyme Assays , Lipase/metabolism , Lipolysis , Phylogeny , Plasmids/genetics , Sequence Homology, Amino AcidABSTRACT
The generation of bradykinin (BK; Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) in blood and kallidin (Lys-BK) in tissues by the action of the kallikrein-kinin system has received little attention in non-mammalian vertebrates. In mammals, kallidin can be generated by the coronary endothelium and myocytes in response to ischemia, mediating cardioprotective events. The plasma of birds lacks two key components of the kallikrein-kinin system: the low molecular weight kininogen and a prekallikrein activator analogous to mammalian factor XII, but treatment with bovine plasma kallikrein generates ornitho-kinin [Thr6,Leu8]-BK. The possible cardioprotective effect of ornitho-kinin infusion was investigated in an anesthetized, open-chest chicken model of acute coronary occlusion. A branch of the left main coronary artery was reversibly ligated to produce ischemia followed by reperfusion, after which the degree of myocardial necrosis (infarct size as a percent of area at risk) was assessed by tetrazolium staining. The iv injection of a low dose of ornitho-kinin (4 microg/kg) reduced mean arterial pressure from 88 +/- 12 to 42 +/- 7 mmHg and increased heart rate from 335 +/- 38 to 402 +/- 45 bpm (N = 5). The size of the infarct was reduced by pretreatment with ornitho-kinin (500 microg/kg infused over a period of 5 min) from 35 +/- 3 to 10 +/- 2% of the area at risk. These results suggest that the physiological role of the kallikrein-kinin system is preserved in this animal model in spite of the absence of two key components, i.e., low molecular weight kininogen and factor XII.
Subject(s)
Bradykinin/analogs & derivatives , Cardiotonic Agents/therapeutic use , Kinins/drug effects , Myocardial Infarction/prevention & control , Vasodilator Agents/therapeutic use , Acute Disease , Angiotensin-Converting Enzyme Inhibitors/pharmacology , Animals , Blood Pressure/drug effects , Bradykinin/therapeutic use , Captopril/pharmacology , Chickens , Disease Models, Animal , Ischemic Preconditioning, Myocardial , Kinins/blood , Kinins/physiology , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Preoperative Care , Vascular Resistance/drug effectsABSTRACT
The generation of bradykinin (BK; Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) in blood and kallidin (Lys-BK) in tissues by the action of the kallikrein-kinin system has received little attention in non-mammalian vertebrates. In mammals, kallidin can be generated by the coronary endothelium and myocytes in response to ischemia, mediating cardioprotective events. The plasma of birds lacks two key components of the kallikrein-kinin system: the low molecular weight kininogen and a prekallikrein activator analogous to mammalian factor XII, but treatment with bovine plasma kallikrein generates ornitho-kinin [Thr6,Leu8]-BK. The possible cardioprotective effect of ornitho-kinin infusion was investigated in an anesthetized, open-chest chicken model of acute coronary occlusion. A branch of the left main coronary artery was reversibly ligated to produce ischemia followed by reperfusion, after which the degree of myocardial necrosis (infarct size as a percent of area at risk) was assessed by tetrazolium staining. The iv injection of a low dose of ornitho-kinin (4 µg/kg) reduced mean arterial pressure from 88 ± 12 to 42 ± 7 mmHg and increased heart rate from 335 ± 38 to 402 ± 45 bpm (N = 5). The size of the infarct was reduced by pretreatment with ornitho-kinin (500 µg/kg infused over a period of 5 min) from 35 ± 3 to 10 ± 2 percent of the area at risk. These results suggest that the physiological role of the kallikrein-kinin system is preserved in this animal model in spite of the absence of two key components, i.e., low molecular weight kininogen and factor XII.
Subject(s)
Animals , Bradykinin/analogs & derivatives , Cardiotonic Agents/therapeutic use , Kinins/drug effects , Myocardial Infarction/prevention & control , Vasodilator Agents/therapeutic use , Acute Disease , Angiotensin-Converting Enzyme Inhibitors/pharmacology , Blood Pressure/drug effects , Bradykinin/therapeutic use , Chickens , Captopril/pharmacology , Disease Models, Animal , Ischemic Preconditioning, Myocardial , Kinins/blood , Kinins/physiology , Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Preoperative Care , Vascular Resistance/drug effectsABSTRACT
BACKGROUND AND STUDY AIM: Small-bowel tumors account for 1% - 3% of all gastrointestinal neoplasms. Recent studies with video capsule endoscopy (VCE) suggest that the frequency of these tumors may be substantially higher than previously reported. The aim of the study was to evaluate the frequency, clinical presentation, diagnostic/therapeutic work-up, and endoscopic appearance of small-bowel tumors in a large population of patients undergoing VCE. PATIENTS AND METHODS: Identification by a questionnaire of patients with VCE findings suggesting small-bowel tumors and histological confirmation of the neoplasm seen in 29 centers of 10 European Countries. RESULTS: Of 5129 patients undergoing VCE, 124 (2.4%) had small-bowel tumors (112 primary, 12 metastatic). Among these patients, indications for VCE were: obscure gastrointestinal bleeding (108 patients), abdominal pain (9), search for primary neoplasm (6), diarrhea with malabsorption (1). The main primary small-bowel tumor type was gastrointestinal stromal tumor (GIST) (32%) followed by adenocarcinoma (20%) and carcinoid (15%); 66% of secondary small-bowel tumors were melanomas. Of the tumors, 80.6% were identified solely on the basis of VCE findings. 55 patients underwent VCE as the third procedure after negative bidirectional endoscopy. The lesions were single in 89.5% of cases, and multiple in 10.5%. Retention of the capsule occurred in 9.8% of patients with small-bowel tumors. After VCE, 54/124 patients underwent 57 other examinations before treatment; in these patients enteroscopy, when performed, showed a high diagnostic yield. Treatment was surgery in 95% of cases. CONCLUSIONS: Our data suggest that VCE detects small-bowel tumors in a small proportion of patients undergoing this examination, but the early use of this tool can shorten the diagnostic work-up and influence the subsequent management of these patients.
Subject(s)
Capsule Endoscopy/methods , Intestinal Neoplasms/diagnosis , Intestinal Neoplasms/epidemiology , Intestine, Small/pathology , Adult , Age Distribution , Aged , Biopsy, Needle , Capsule Endoscopy/adverse effects , Early Diagnosis , Europe/epidemiology , Female , Humans , Incidence , Male , Middle Aged , Multicenter Studies as Topic , Neoplasm Staging , Normal Distribution , Probability , Risk Assessment , Sensitivity and Specificity , Sex Distribution , Statistics, NonparametricABSTRACT
OBJECTIVE: To assess the distribution, prevalence and severity of malocclusion and orthodontic treatment needs in schoolchildren from the northeast of Brazil aged between 13 and 15 years. RESEARCH DESIGN: Cross-sectional study. PARTICIPANTS: A sample of 600 adolescents (264 males and 336 females) randomly selected and representative of schoolchildren living in Recife (Brazil) was obtained from 12 public schools. METHOD: The need for orthodontic treatment was measured using the Dental Aesthetic Index (DAI). RESULTS: Most of the subjects (77%) were deemed to require orthodontic treatment. Only about 5.8% had a handicapping malocclusion that needed mandatory treatment. A severe malocclusion for which treatment was highly desirable was recorded in 47.5% of the adolescents and 23.7% had a definite malocclusion for which treatment was elective. Three main occlusal features were responsible for allocating subjects into the group of "orthodontic treatment required": crowding (47.3%), tooth loss (22.3%) and maxillary overjet of more than 3 mm (21.8%). There were no significant differences (p > 0.05) in mean DAI scores between males and females. CONCLUSIONS: 77% of adolescents from northeast Brazil were in need of orthodontic treatment for dental health reasons. The distribution of DAI scores among Brazilian adolescents is different from that reported in other populations. This study provides baseline data on the need and demand for orthodontic treatment among Brazilian students.