Endothelial cells and angiogenesis in the horse in health and disease—A review

The cardiovascular system is the first functional organ in the embryo, and its blood vessels form a widespread conductive network within the organism. Blood vessels develop de novo, by the differentiation of endothelial progenitor cells (vasculogenesis) or by angiogenesis, which is the formation of new blood vessels from existing ones. This review presents an overview of the current knowledge on physiological and pathological angiogenesis in the horse including studies on equine endothelial cells. Principal study fields in equine angiogenesis research were identified: equine endothelial progenitor cells; equine endothelial cells and angiogenesis (heterogeneity, markers and assessment); endothelial regulatory molecules in equine angiogenesis; angiogenesis research in equine reproduction (ovary, uterus, placenta and conceptus, testis); angiogenesis research in pathological conditions (tumours, ocular pathologies, equine wound healing, musculoskeletal system and laminitis). The review also includes a table that summarizes in vitro studies on equine endothelial cells, either describing the isolation procedure or using previously isolated endothelial cells. A particular challenge of the review was that results published are fragmentary and sometimes even contradictory, raising more questions than they answer. In conclusion, angiogenesis is a major factor in several diseases frequently occurring in horses, but relatively few studies focus on angiogenesis in the horse. The challenge for the future is therefore to continue exploring new therapeutic angiogenesis strategies for horses to fill in the missing pieces of the puzzle.     

Kinetics and role of antibodies against intimin β in colostrum and in serum from goat kids and longitudinal study of attaching and effacing Escherichia coli in goat kids

The presence of antibodies to the intimin β-binding region (Int280-β) of attaching and effacing Escherichia coli (AEEC) in serum from 20 goat kids from 2 herds, as well as in goat colostrum, was investigated by enzyme-linked immunosorbent assay. In addition, the onset and subsequent pattern of shedding of AEEC from the same goat kids over a 6-mo period was investigated. All the colostrum and serum samples tested contained antibodies against Int280-β. The association between the antibody titer and the isolation of AEEC suggests that antibodies to intimin β do not prevent colonization of the intestine by AEEC in goat kids. The AEEC were generally shed only transiently. Most AEEC isolated from the kids belonged to serogroup O26. Three isolates belonged to serogroup O157. These data show that goat kids may be a reservoir of AEEC that are potentially pathogenic for humans.     

Toxoplasma gondii and Neospora caninum in wildlife: common parasites in Belgian foxes and Cervidae?

Sera from Cervidae were tested for the presence of antibodies against Neospora caninum using ELISA; and against Toxoplasma gondii using SAG1-ELISA and a commercially available agglutination test. The T. gondii seroprevalence was 52% (38/73) in roe deer (Capreolus capreolus), 0% in bred fallow deer (0/4) (Dama dama) and red deer (0/7) (Cervus elaphus). We found 2.7% of the roe deer samples and none of the bred deer samples positive for N. caninum. Brain samples from wild roe deer, red deer and red foxes (Vulpes vulpes) were tested for the presence of T. gondii and N. caninum DNA using multiplex real-time PCR. We detected T. gondii in 18.8% (57/304) of the red foxes and in 1 of the 33 deer samples. N. caninum was found in 6.6% of the red foxes and in 2 roe deer samples. Twenty-six of the T. gondii positive DNA extracts from the red fox samples were genotyped. Twenty-five were type II and only one was found to be type III.     

Seasonal Changes in Forage Nutrients and Mineral Contents in Water Resources,Forage and Yak Blood

This paper reports results of a study conducted to investigate the concentrations of seven mineral elements in yak blood, forage and water resources around the Qinghai Lake in Qinghai Province in different seasons. Meanwhile, the nutritional compositions of the forage were also surveyed. The results suggest that the mineral elements and the forage nutrients change in a seasonal pattern. In yak blood,the sodium(Na)concentration varies from 0.291 to 0.034 mg/mL,and this is lower than the normal value. In the forage,the ratio calcium(Ca)to phosphorus(P)is 4.06～7.47:1 and potassium(K)to Na 30～27:1. These results indicate that the nutrition of the yak in the area is deficient in Na but high in K. For the withered forage sampled in February,the protein content is only 31.14% of the total protein in the forage growing at puerile stage in June. The severe loss of protein by 68. 9% and decrease of effective nutrients in the wintered forage are considered to be the reasons resulting in the poor condition of yak in winter and spring seasons.     

Expression of hypoxia-inducible factor-1α and vascular density in mammary adenomas and adenocarcinomas in bitches

Background

The study aimed at examining hypoxia-inducible factor (HIF)1α expression in adenocarcinomas and adenomas in bitches in regard to tumour malignancy grade, proliferation, apoptosis and vascularisation. Therefore, paraffin sections of 15 adenomas and 64 adenocarcinomas sampled from 79 dogs aged 6 to 16 years were analysed.

Results

A significantly higher HIF-1α expression was noted in adenocarcinomas in comparison to adenomas (P < 0.0004). Moreover, HIF-1α expression in adenocarcinomas correlated positively with tumour malignancy grade (r = 0.59, P < 0.05), Ki-67 antigen expression (r = 0.43; P < 0.0005), TUNEL-positive cells (r = 0.62, P < 0001) and tumour vascularity measured by quantification of vessels characterized by the expression of von Willebrand Factor (r = 0.57, P < 0.05).

Conclusion

Results of this study indicate a similar biological role of HIF-1α in dogs and in humans, which may confirm suitability of the animal model in investigations on progression of tumours in humans.     

Effects of lycopene on in vitro quality and lipid peroxidation in refrigerated and cryopreserved turkey spermatozoa

1. The effects of lycopene-enriched extenders on the in vitro quality of turkey semen including lipid peroxidation were examined after chilled and frozen storage.

2. Five pools of semen diluted in extenders containing 0, 0·05 or 0·1?mg/ml of lycopene were stored at 5°C for 48?h or cryopreserved as pellets and the following variables determined in fresh samples and samples stored chilled or frozen: sperm motility, viability, osmotic resistance, DNA integrity and lipid peroxidation (as malonaldehyde production).

3. Semen quality was generally compromised after storage, especially post-freezing. However, in the presence of the highest dose of lycopene, both the viability and osmotic-resistance of chilled spermatozoa and the DNA integrity of frozen spermatozoa were similar to those of fresh spermatozoa.

4. Greater lipid peroxidation was detected in refrigerated compared to fresh or cryopreserved spermatozoa. However, spermatozoa chilled in lycopene-enriched extenders showed significantly lower malonaldehyde levels than those chilled without lycopene, while the addition of lycopene to the freezing medium served to maintain the lipid peroxidation levels observed in fresh semen.

5. In conclusion, the presence of lycopene in the extender improved the survival of turkey spermatozoa after liquid-storage and protected DNA integrity against cryodamage. The beneficial effects of lycopene observed could be related to its capacity to diminish sperm lipid peroxidation during refrigeration or cryopreservation.     

Prevalence and Heritability of Symptomatic Syringomyelia in Cavalier King Charles Spaniels and Long‐term Outcome in Symptomatic and Asymptomatic Littermates

Background

Syringomyelia (SM) is common in the Cavalier King Charles Spaniel (CKCS). Dogs with syringes express clinical signs or might be clinically silent.

Objectives

To investigate the prevalence and heritability of symptomatic SM, the association between clinical signs and magnetic resonance imaging (MRI) findings, and long‐term outcome.

Animals

All CKCS registered in the Danish Kennel Club in 2001 (n = 240).

Methods

A cross‐sectional questionnaire‐based prevalence study validated by telephone interviews and clinically investigated clinical signs of SM. Dogs were 6 years at the time of investigation. A prospective observational litter study including clinical investigations, MRI and 5‐year follow‐up of symptomatic and asymptomatic siblings. Heritability was estimated based on the scale of liability in the study population and litter cohort.

Results

The cross‐sectional study estimated a prevalence of symptomatic SM at 15.4% in the population. Thirteen symptomatic and 9 asymptomatic siblings participated in the litter study. Spinal cord syringes were confirmed in 21 of 22 littermates (95%). Syrinx diameter and mean syrinx : spinal cord ratio were significantly correlated with clinical signs (P < .01). Estimated heritability of symptomatic SM was 0.81. Symptomatic SM motivated euthanasia in 20%. Dogs with syringes, which expressed no clinical signs at the age of 6, remained asymptomatic in 14/15 cases (93%).

Conclusions and Clinical Importance

The prevalence of symptomatic SM is high and genetics have a high impact on clinical disease expression. Further investigations of factors influencing the outbreak threshold of clinical signs of SM are desirable.     

Abnormal platelet activity in dogs and cats – impact and measurement

Abnormal platelet activity can either lead to bleeding tendencies or inappropriate thrombus formation and can occur secondarily to a wide variety of disease processes, with a range of clinical consequences and severity. This article will discuss the pathophysiology of platelet function abnormalities and consider a logical diagnostic approach applicable to veterinary practice. Recent advances in platelet function testing will then be discussed, with regards to detection of platelet dysfunction and tailoring of pharmacological manipulation. Although many of these tests are still confined to research or academic institutions, techniques for indirectly assessing platelet function are starting to become more widely available. Although we still require further research to develop guidelines for the use of these tests in clinical decision-making, the recent advances in this field are an exciting step forward in being able to detect and manage platelet dysfunction in both primary care and referral practice.     

β-Adrenoceptors in Equine Trachea and Heart

The density and subtype pattern of -adrenoceptors in equine tracheal epithelium, tracheal smooth muscle and heart from 6–9 horses were investigated by radioligand binding studies using the non-selective -adrenoceptor antagonist 125I-cyanopindolol (ICYP). The specific binding of ICYP was 341±162 fmol/mg protein (mean±SD) for epithelium, 42±13 fmol/mg for smooth muscle and 124±39 and 101±19 fmol/mg for the cardiac atrium and ventricle, respectively. The Kd value of ICYP was 6.7–10.2 pmol/L.In competition studies, different concentrations of either the 2-selective drug ICI 118551 or the 1-selective CGP 20712A competed with 25 pmol/L ICYP for the binding sites. The competition curves for tracheal smooth muscle and epithelium were monophasic with an approximate Kd value for ICI 118551 of 1 nmol/L and for CGP 20712A of 10 000 nmol/L. This corresponds to known Kd values for these substances binding to 2-adrenoceptors. 2-Adrenoceptors were also found in the heart, most pronounced in the atrium, where the density was 29%±6% (mean±SD) of the total receptor density. CGP 20712A and ICI 118551 bound to the dominating binding site of 1-adrenoceptors in the heart with Kd values of approximately 1 nmol/L and 100 nmol/L, respectively.     

Canine microglial cells: stereotypy in immunophenotype and specificity in function?

Microglial cells represent the endogenous immune system of the central nervous system (CNS). Upon pathological insults they reveal their immunological potential aimed at regaining homeostasis. These reactions have long been believed to follow a uniform and unspecific pattern which is irrespective to the underlying disease entity. Evidence is growing that this view seriously underrates microglial competence as the defenders of the CNS. In the present study, microglial cells of 47 dogs were examined ex vivo by means of flow cytometry. Ex vivo examination included immunophenotypic characterization using eight different surface markers and functional studies such as phagocytosis assay and the reactive oxygen species (ROS) generation test. The dogs were classified according to their histopathological diagnoses in disease categories (controls, canine distemper virus (CDV) induced demyelination, other diseases of the CNS) and results of microglial reaction profiles were compared. Immunophenotypic characterization generally revealed relative high conformity in the microglial disease response among the different groups, however the functional response was shown to be more specific. Dogs with intracranial inflammation and dogs with demyelination showed an enhanced phagocytosis, whereas a significant up-regulation of ROS generation was found in dogs with demyelination due to CDV infection. This strongly suggests a specific response of microglia to infection with CDV in the settings of our study and underlines the pivotal role of microglial ROS generation in the pathogenesis of demyelinating diseases, such as canine distemper.     

Variation in mammal species richness and relative abundance in the Karoo§

Understanding how climate change and land transformation may impact the distribution and diversity of wildlife species requires landscape-level foundational biodiversity surveys. The Karoo BioGaps Project aims to provide such data and to support the scientific assessment for shale gas development projects in the Karoo basin. In this paper we present the findings of the BioGaps mammal survey, which recorded medium and large mammals across twenty-five 1 km × 1 km sampling sites within the proposed fracking footprint using camera trapping techniques. We use sample rarefaction curves, non-parametric species richness estimators and non-metric multidimensional scaling plots to explore both species richness and community structure. We also used a generalised linear model to investigate how species diversity varies with both site-specific and landscape-level predictors. A total of 38 species were recorded at the majority of sites. Longitude (z = 4.018, p = 0.0005) emerged as the best predictor of species diversity across the study area, which we suggest is linked to the clear east–west aridity gradient. Together these results reveal the cosmopolitan distribution of the mammal taxa in the Karoo and could be used to inform decision-making linked to mining activities in the area.     

Detection and molecular characterization of porcine kobuvirus in piglets in 2009–2013 in northern Thailand

Tropical Animal Health and Production - A total of 636 fecal samples collected from piglets with and without diarrhea during 2009 to 2013 were tested for porcine kobuvirus by RT-PCR. From a total...     

α- and β-adrenergic influences on metabolism in heart and skeletal muscle

Using fenylephrine, isoprenaline, noradrenaline, adrenaline and receptor blockers, adrenergic regulation of metabolism in working heart as well as in skeletal resting muscle of rats was investigated. Changes were determined in: phosphocreatine level and³²P incorporation into phosphocreatine; phosphorolytic and hydrolytic activities of glycogenolysis; glycogen synthetase activity and¹⁴C-glucose incorporation into glycogen; and the level of cAMP. It was found that, contrary to the β-receptor, stimulation of the α-adrenergic receptor inhibits catabolism of macroenergetic phosphates and diminishes glycogen decomposition on the phosphorolytic pathway while increasing the hydrolysis of glycogen and glycogen synthetase activity as well as glucose incorporation into glycogen. On the basis of these data a hypothesis may be put forward that the influence of the α-adrenergic receptor results in the diminishing of energy utilization and in the activation of glycogen anabolism in heart and skeletal muscle. An adrenergic intrasystemic receptor antagonism in the regulation of energy consumption by heart and skeletal muscle is indicated. The α-receptor seems to be an adrenergic intrasystemic moderator of the metabolism rate which is accelerated by β-receptor influence. This suggests that besides sympathetico-parasympathetic intersystemic antagonism regulating the heart work and thereby cardiac energy consumption, there is a separate adrenergic mechanism directly controlling the balance between catabolism and anabolism in heart muscle. This mechanism, based on α- and β-adrenergic receptor antagonism, also controls the metabolic balance in skeletal muscles.     

Inhibin-α immunohistochemical expression in mature and immature canine Sertoli and Leydig cells

Formalin-fixed, paraffin-embedded sections from 32 canine pairs of testes were immunohistochemically examined for Inhibin-α (INHα). Samples were subdivided into two groups (group 1, neonates; group 2, puppies and adults) and results statistically compared. Inhibin-α was significantly expressed only in Sertoli cells of neonatal testes, while in Leydig cells it was expressed without significant difference between groups. These results suggest that, in dogs, INHα expression switches from Sertoli to Leydig cells during testicular maturation and that, in adult, Leydig cells represent the main source of INHα.     

Pre-implantation exogenous progesterone and pregnancy in sheep. Ⅱ. Effects on fetalplacental development and nutrient transporters in late pregnancy

Background: Administration of progesterone(P4) to ewes during the first 9 to 12 days of pregnancy accelerates blastocyst development by day 12 of pregnancy, likely due to P4-induced up-regulation of key genes in uterine epithelia responsible for secretion and transport of components of histotroph into the uterine lumen. This study determined if acceleration of blastocyst development induced by exogenous P4 during the pre-implantation period affects fetal-placental development on day 125 of pregnancy. Suffolk ewes(n = 35) were mated to fertile rams and assigned randomly to receive daily intramuscular injections of either corn oil vehicle(CO, n = 18) or 25 mg progesterone in CO(P4, n = 17) for the first 8 days of pregnancy. All ewes were hysterectomized on day 125 of pregnancy and: 1) fetal and placental weights and measurements were recorded; 2) endometrial and placental tissues were analyzed for the expression of candidate mRNAs involved in nutrient transport and arginine metabolism; and 3) maternal plasma, fetal plasma, allantoic fluid, and amniotic fluid were analyzed for amino acids,agmatine, polyamines, glucose, and fructose.Results: Treatment of ewes with exogenous P4 did not alter fetal or placental growth, but increased amounts of aspartate and arginine in allantoic fluid and amniotic fluid, respectively. Ewes that received exogenous P4 had greater expression of mRNAs for SLC7 A1, SLC7 A2, SLC2 A1, AGMAT, and ODC1 in endometria, as well as SLC1 A4, SLC2 A5, SLC2 A8 and ODC1 in placentomes. In addition, AZIN2 protein was immunolocalized to uterine luminal and glandular epithelia in P4-treated ewes, whereas AZIN2 localized only to uterine luminal epithelia in CO-treated ewes.Conclusions: This study revealed that exogenous P4 administered in early pregnancy influenced expression of selected genes for nutrient transporters and the expression of a protein involved in polyamine synthesis on day 125 of pregnancy, suggesting a ‘programming' effect of P4 on gene expression that affected the composition of nutrients in fetal-placental fluids.     

Acute changes in the concentrations of prostaglandin F2α (PGF) and cortisol in uterine and ovarian venous blood during PGF-induced luteolysis in cows

Prostaglandin F2α (PGF) is considered to be the main luteolysin in cattle. We have previously demonstrated that cortisol (Cr) suppresses PGF production in non-pregnant bovine endometrium. This study was carried out to test whether exogenous PGF increases ovarian and/or uterine PGF production and to determine the temporal relationship between PGF and Cr in ovarian and uterine circulations during PGF-induced luteolysis in cows. Catheters were inserted into the ovarian vein (OV), uterine vein (UV) and jugular vein (JV) of 10 cows on Day 9 of the oestrous cycle (Ovulation = Day 0) for frequent blood collection. On Day 10, the cows were divided randomly into two groups and treated with a luteolytic dose of a PGF analogue (cloprostenol) or saline solution. Blood samples were collected at -0.25, 0, 0.25, 0.5, 1 and 2 h and then at 2-h intervals until 12 h after treatment (0 h). The basal concentrations of PGF and Cr in OV and UV plasma were not significantly different. Injection of a PGF analogue induced more than twofold increases in the levels of PGF between 0.25 and 1 h in UV plasma, but not in OV plasma. PGF increased (p < 0.05) the concentrations of Cr in OV, UV and JV plasma between 0.5 and 1 h. The Cr levels in OV, UV and JV plasma were similar. The PGF levels in UV plasma decreased after Cr reached its highest levels. The overall results suggest that the uterus rather than the ovary increases PGF production in response to PGF injection. Based on the temporal changes of PGF and Cr in the ovarian and uterine circulations, Cr may act to reduce uterine PGF production in non-pregnant cows in vivo.