Relationships between haptoglobin and inter-alpha trypsin inhibitor heavy chain 4 levels in local and peripheral blood and systemic inflammatory response syndrome in bitches with pyometra.

We aimed to assess the usefulness of monitoring inter-alpha trypsin inhibitor heavy chain 4 (ITIH4) and haptoglobin (Hp) in peripheral and local blood in canine pyometra, and evaluation the relationships among acute phase proteins (APPs), systemic inflammatory response syndrome (SIRS) and the presence of bacteria. The material was collected from bitches with pyometra and from healthy ones. Blood was taken from the cephalic and uterine veins. APPs levels were quantified by ELISA. In the peripheral circulation, the Hp was higher in animals with open-cervix pyometra (OCP) than in the closed-cervix pyometra (CCP) and the control group. The Hp concentration was not correlated with age, with the presence of SIRS or with the type of bacteria (Gram-negative, Gram-positive or mixed flora). The ITIH4 concentrations in the peripheral blood did not differ significantly in the cases of pyometra. The Hp concentration in the local circulation increased in the OCP but not in the CCP groups, although the histopathological changes in the endometrium were similar. Peripheral Hp concentrations may be a useful tool in differentiating between the types of pyometra.

Impact of Chlorogenic Acid on Peripheral Blood Mononuclear Cell Proliferation, Oxidative Stress, and Inflammatory Responses in Racehorses during Exercise.

Green coffee extract is currently of great interest to researchers due to its high concentration of chlorogenic acid (CGA) and its potential health benefits. CGA constitutes 6 to 10% of the dry weight of the extract and, due to its anti-inflammatory properties, is a promising natural supplement and agent with therapeutic applications. The purpose of our study was to discover the effects of CGA on peripheral blood mononuclear cell proliferation, and the production of pro- and anti-inflammatory cytokines as well as reactive oxidative species (ROS) in horses during exercise. According to the findings, CGA can affect the proliferation of T helper cells. In addition, at a dose of 50 g/mL, CGA increased the activation of CD4+FoxP3+ and CD8+FoxP3+ regulatory cells. Physical activity decreases ROS production in CD5+ monocytes, but this effect depends on the concentration of CGA, and the effect of exercise on oxidative stress was lower in CD14+ than in CD5+ cells. Regardless of CGA content, CGA significantly increased the release of the anti-inflammatory cytokine IL-10. Moreover, the production of IL-17 was greater in cells treated with 50 g/mL of CGA from beginners compared to the control and advanced groups of horses. Our findings suggest that CGA may have immune-enhancing properties. This opens new avenues of research into the mechanisms of action of CGA and possible applications in prevention and health promotion in sport animals.

Exercise-related changes in the anabolic index (testosterone to cortisol ratio) and serum amyloid A concentration in endurance and racehorses at different fitness levels.

Increased training loads in both human and equine athletes are generally implemented to improve their physical performance. These loads are tolerated only within appropriate training periodization with regard to recovery time. Otherwise, training overload causes failure in the systemic adaptation, which at first leads to overreaching, and progressively to overtraining syndrome (OTS). Exercise endocrinology, and anabolic/catabolic balance as an indicator of athlete performance status and OTS has continued to attract attention. In human medicine, changes in testosterone and cortisol levels, as well as the testosterone to cortisol ratio (T/C; anabolic index), are suggested to be sensitive stress markers. However, there is a lack of research investigating these parameters for use in equine sports medicine. The aim of the study was to investigate the differences in testosterone, cortisol, and T/C in response to a single training session in two types of equine sports: endurance and race, together with serum amyloid A (SAA), the main acute phase response indicator of physical effort, and the overall health status in horses. Two groups of horses were enrolled in the study: endurance (n = 12) and racehorses (n = 32) of different fitness level. Blood samples were obtained before and after the exercise. On average, T increased 2.5 times after the race training in experienced racehorses and dropped in endurance horses regardless the fitness level (p < 0.05). In endurance horses, a decrease in T/C occurred after training in inexperienced horses (p < 0.05). In racehorses, a T/C decrease occurred in the inexperienced group (p < 0.05) and an increase in the experienced (p < 0.01). In conclusion, T/C ratio was found to be a potentially reliable indicator of fitness status especially in racing horses. These findings provide insight into the physiological response of the horses to different types of exercise and the potential use of hormone levels as markers of performance and adaptation.

Acid-Base, Gas, Ions, and Glucose Analysis in Follicular Fluid in Holstein-Friesian Dairy Cows Is Associated with the Follicle Size in Poland.

The lack of fertilization and early pregnancy loss is seen in the quality and maturation of the oocytes. The environment of the first divisions and maturation of the oogonia, which is also a reflection of the quality of the oocyte, is the follicular fluid (FF). Thus, the purpose of this study was to investigate the variations in parameters such as pH, pCO2, pO2, standard HCO3-, actual HCO3-, base excess (BE), base excess of extracellular fluid (BE ecf), ctCO2, ions: Na+, K+, actual ionized calcium Ca2+, adjusted ionized calcium (at pH 7.4) Ca2+ (7.4), Cl-, anion gap (AnGap), and glucose in FF collected from different size follicles in dairy cattle. The most apparent differences were related to pH, K+, and Ca2+ 7.4 concentration in comparison to changes in follicle size (p < 0.05). Several trends were also evaluated as an increase in follicular size was followed by an increase in pH, BE, and Ca2+ 7.4 value and a decrease in the K+ concentration (p < 0.05). In conclusion, there are changes in FF formularies associated with the size of follicles. However, more research is necessary to establish the reference value, which then could be one of the factors describing the quality of the follicle and the developmental potential of the associated oocyte.

Exercise-induced Haematological and Blood Lactate Changes in Whippets Training for Lure Coursing.

Introduction: Whippets are traditionally trained to compete in lure coursing. While in humans and horses, training is routinely monitored by special tests, this is not carried out in the training of whippets. The aim of this study was to check if laboratory tests designed for racehorses could be useful in monitoring whippets training for lure coursing. Material and Methods: Blood samples were taken from 14 whippets at several time points: before exercise (including warm-up), immediately after, 15 min after and 30 min after exercise sessions of straight 400 m runs (T) and coursing (C). Routine haematological values and lactate concentrations (LA) were measured. Results: White blood cell count, red blood cell count, haemoglobin concentration and haematocrit increased significantly in both types of exertion, and no differences between the types were observed. The LA measured immediately after the run were increased, but there was no significant difference between the types of session (T and C). After both types of activity, LA decreased within 30 min post run by 9-11 mmol/L. Lactate concentrations were significantly higher 30 min after the T sessions than after the C sessions. Conclusion: The results confirmed that typical exercise-induced changes occurred in whippets training for lure coursing; however, the scale of changes was different to that in horses. The sampling scheme used in racehorses can be applied to whippets and can be useful as a laboratory tool for monitoring their training.

How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females (Cervus elaphus L.) in Different Reproductive Stages?

Reproductive and condition parameters' dependency on immune status in seasonally reproducing ruminants such as red deer have not been outlined to date. We determined T and B blood lymphocytes; the concentration of IgG, cAMP, haptoglobulin, and 6-keto-PGF1α in blood plasma; and the mRNA and protein expression of PG endoperoxide synthase 2, 5-lipoxygenase, PGE2 synthase (PGES), PGF2α synthase (PGFS), PGI2 synthase (PGIS), leukotriene (LT)A4 hydrolase, and LTC4 synthase (LTC4S) in the uterine endo- and myometrium, on the 4th (N = 7) and 13th (N = 8) days of the estrous cycle, in anestrus (N = 6) and pregnancy (N = 8) in hinds. An increase in CD4+ T regulatory lymphocyte percentage during the estrous cycle and anestrus compared with pregnancy was recorded; the opposite effect was observed for CD21+ B cells (p < 0.05). cAMP and haptoglobin concentration were elevated during the cycle, as was IgG on the fourth day of the cycle, whereas 6-keto-PGF1α concentration was the highest in pregnancy, and the nearest in anestrus similarly were LTC4S, PGES, PGFS, and PGIS protein expression in the endometrium (p < 0.05). We showed an interaction between the immune system activation and AA-metabolite production in the uterus throughout different reproductive stages. IgG, cAMP, haptoglobin, and 6-keto-PGF1α concentrations are valuable candidates for markers of reproductive status in hinds. The results help expand our knowledge of the mechanisms underlying seasonal reproduction in ruminants.

Comparison of blood gas parameters, ions, and glucose concentration in polish Holstein-Friesian Dairy cows at different milk production levels.

Genetic selection for increased milk yield has been a key driver of dairy intensification. The modern dairy cow produces much higher amounts of milk than the cattle of several years ago, and this may have an influence on hematologic values at different stages of lactation and on cows with different levels of milk production. The purpose of the study was to investigate the variations in blood parameters such as Ht, tHb, sO2, FO2Hb, FCOHb, FMetHb, FHHb, pH, pCO2, pO2, standard HCO3-, actual HCO3-, BE, BE ecf, ctCO2, BO2, p50, and ctO2 in cows at different milk production levels. In addition, ions such as Na+, K+ , Ca++, Ca++ (7.4), and Cl-, and AnGap and glucose were examined. Our findings indicated that differences in the examined blood parameters between low and high-production dairy cattle do exist. The most apparent differences were connected with blood pH (p < 0.01), oxygen metabolism (Ht, tHb, sO2, FO2Hb; p < 0.01), and glucose utilization (p < 0.01) The results confirm that the parameters connected with blood oxygen metabolism and glucose metabolism increase significantly in high-production animals. In conclusion, reference values should be considered in light of the lactation stage and level of milk production, because these might influence how changes should be interpreted. The main limitation of the study is the delay to analysis. However, the blood was properly stored (4C), thus changes were delayed. Anyway, it is very hard in the field practice to perform it within 5 min after the blood collection and according to studies it has low impact on clinical outcomes.

Changes in novel anti-infalmmatory cytokine concetration in the bood of endurance and race horses at different levels of training.

Several anti-inflammatory cytokines have been proposed as markers for exercise monitoring in humans such as the interleukin 1 receptor agonist (IL-ra), or interleukin 13 (IL-13). Equine athletes may be considered a model for human exercise physiology research, however there is a lack of such studies of this species. Thus, we decided to examine the changes of IL-1ra and IL-13 in serum concentration during aerobic (endurance) and anaerobic (race) exercise in horses of different fitness levels in comparison with the well-known anti-inflammatory cytokine interleukin 10 (IL-10). The group of endurance horses (n = 13) consisted of animals competing over 100 (n = 7) and 120 km (n = 6) rides. The group of racehorses (n = 18) consisted of trained (n = 9) and untrained (n = 9) animals. The blood samples were obtained before and after the exercise. The ELISA test was performed to evaluate the changes of IL-1ra, IL-13 and IL-10 during different types of exercise. In endurance horses there was an increase in IL-13 (p = 0.0012) after the 100 km ride and in IL-1ra (p = 0.0411) after the 120 km ride. In race horses there was a higher IL-13 basal serum concentration in the untrained group, as well as a decrease of IL-13 after exercise (p = 0.0188). In trained racehorses there was an increase in IL-1ra (p < 0.0001) and IL-13 after exercise (p = 0.0028). In conclusion, the reaction of IL-1ra and IL-13 to different types of exercise differ from each other. Thus, in future, they may be helpful in monitoring the fitness of horses, however more research is needed.

C-Reactive Protein as a Diagnostic Marker in Dogs: A Review.

Acute phase response is a nonspecific reaction to disturbances in homeostasis during which the production of some Acute Phase Proteins (APPs) is stimulated; they are sensitive but nonspecific markers of systemic inflammatory processes. The major positive APP in dogs is the C-reactive protein (CRP). The dynamic of its concentration changes fast, rising and decreasing rapidly with the onset and removal of the inflammatory stimulus. It increases within the first 4-24 h after the stimulus and reaches up to a 50-100-fold increase of the baseline level. It has been documented that this APP's concentration is elevated during several diseases, such as pyometra, panniculitis, acute pancreatitis, polyarthritis, sepsis, immune-mediated hemolytic anemia, and neoplasia in dogs. In clinical practice, canine CRP is mostly measured to detect and monitor systemic inflammatory activity and the efficacy of treatments, because it is a more sensitive marker than shifts in leukocyte counts. Blood serum CRP concentration is becoming a part of routine biochemistry panels in many countries. In this article, changes in CRP concentration and its clinical application in healthy and diseased dogs are discussed.

The Novel Diagnostic Techniques and Biomarkers of Canine Mammary Tumors.

Canine mammary tumors (CMTs) are considered a serious clinical problem in older bitches. Due to the high malignancy rate and poor prognosis, an early diagnosis is essential. This article is a summary of novel diagnostic techniques as well as the main biomarkers of CMTs. So far, CMTs are detected only when changes in mammary glands are clinically visible and surgical removal of the mass is the only recommended treatment. Proper diagnostics of CMT is especially important as they represent a very diverse group of tumors and therefore different treatment approaches may be required. Recently, new diagnostic options appeared, like a new cytological grading system of CMTs or B-mode ultrasound, the Doppler technique, contrast-enhanced ultrasound, and real-time elastography, which may be useful in pre-surgical evaluation. However, in order to detect malignancies before macroscopic changes are visible, evaluation of serum and tissue biomarkers should be considered. Among them, we distinguish markers of the cell cycle, proliferation, apoptosis, metastatic potential and prognosis, hormone receptors, inflammatory and more recent: metabolomic, gene expression, miRNA, and transcriptome sequencing markers. The use of a couple of the above-mentioned markers together seems to be the most useful for the early diagnosis of neoplastic diseases as well as to evaluate response to treatment, presence of tumor progression, or further prognosis. Molecular aspects of tumors seem to be crucial for proper understanding of tumorigenesis and the application of individual treatment options.

Reduced Graphene Oxide Modulates the FAK-Dependent Signaling Pathway in Glioblastoma Multiforme Cells In Vitro.

Aggressive invasiveness is a common feature of malignant gliomas, despite their high level of tumor heterogeneity and possible diverse cell origins. Therefore, it is important to explore new therapeutic methods. In this study, we evaluated and compared the effects of graphene (GN) and reduced graphene oxides (rGOs) on a highly invasive and neoplastic cell line, U87. The surface functional groups of the GN and rGO flakes were characterized by X-ray photoelectron spectroscopy. The antitumor activity of these flakes was obtained by using the neutral red assay and their anti-migratory activity was determined using the wound healing assay. Further, we investigated the mRNA and protein expression levels of important cell adhesion molecules involved in migration and invasiveness. The rGO flakes, particularly rGO/ATS and rGO/TUD, were found highly toxic. The migration potential of both U87 and Hs5 cells decreased, especially after rGO/TUD treatment. A post-treatment decrease in mobility and FAK expression was observed in U87 cells treated with rGO/ATS and rGO/TUD flakes. The rGO/TUD treatment also reduced ß-catenin expression in U87 cells. Our results suggest that rGO flakes reduce the migration and invasiveness of U87 tumor cells and can, thus, be used as potential antitumor agents.

Canine B Cell Lymphoma- and Leukemia-Derived Extracellular Vesicles Moderate Differentiation and Cytokine Production of T and B Cells In Vitro.

Extracellular vesicles (EVs) are formed in physiological and pathological conditions by almost all mammalian cells. They are known as submicron "molecules" that transport and horizontally transfer their cargo from maternal cells to donor cells. Moreover, cancer cells produce tumor-derived EVs (TEVs), which are present in blood of patients with solid tumors and those with hematological malignancies. Their role in evading immune system surveillance and induction of immunosuppression in hematological cancer is limited. According to the authors' best knowledge, there is no information about the impact of TEVs from canine lymphoma (CLBL-1) and leukemia (CLB70) on lymphocytes isolated from peripheral blood mononuclear cells (PBMCs). In conclusion, we demonstrate in in vitro experiments that CLBL-1 EVs and CLB70 EVs are effectively taken up by T and B lymphocytes. TEVs decrease the percentage of B lymphocytes and increase that of T lymphocytes, and change T cells' phenotype into the effector memory (EM) or terminally differentiated effector memory (TEMRA) subtype after in vitro co-culturing. Moreover, CLBL70 EVs have pro-tumorogenic properties by inhibiting the production of CD8+IL-17+ cells.

MHCII Expression on Peripheral Blood Monocytes in Canine Lymphoma: An Impact of Glucocorticoids.

An increase in the percentage of monocytes with reduced HLA-DR expression and immunosuppressive properties has been reported in numerous human neoplastic diseases, including lymphoma. However, there are no analogous studies on phenotypical variations in the peripheral blood monocytes in dogs with lymphoma. The aim of this study was to determine the difference in the expression of the MHCII molecule on peripheral blood monocytes in dogs with lymphoma before any treatment (NRG) and in dogs that had previously received glucocorticoids (RG) in comparison to healthy dogs. Flow cytometry immunophenotyping of peripheral blood leukocytes was performed using canine-specific or cross-reactive antibodies against CD11b, CD14 and MHCII. In the blood of dogs with lymphoma (NRG and RG), compared to that of healthy ones, the MHCII+ and MHCII- monocytes ratio was changed due to an increase in the percentage of MHCII- monocytes. The number of MHCII- monocytes was significantly higher only in RG dogs compared to healthy ones, which might result from the release of these cells from the blood marginal pool due to the action of glucocorticoids. Our results encourage further studies to assess if changes in MHCII expression affect immune status in dogs with lymphoma.

The many problems of somatic cell nuclear transfer in reproductive cloning of mammals.

In 1996, when Dolly the sheep was born, a new, utopian era was expected to begin. Science fiction and popular culture instantly threatened the public with shortly upcoming human clones, portraying it as a very easy and instant procedure. Practice has proven otherwise; it exposed how little is known about the early development of mammals and epigenetic reprogramming. Unfortunately, somatic cell nuclear transfer success rate in mammals has not changed much since its very beginning. It is not uncommon that hundreds of oocytes need to be reconstructed to obtain a single live birth. In this review we provide a brief summary of the progress and problems of the field; beginning with selection of the donor cells and their susceptibility to different methods of epigenetic reprogramming; methods of the later gene activation, placental abnormalities, and their possible causes; to health issues that such offspring is prone to.

Platelet Extracellular Vesicles Are Taken up by Canine T Lymphocytes but Do Not Play a Role in Their Proliferation, Differentiation and Cytokine Production In Vitro.

Eukaryotic and prokaryotic cells in physiological and pathological conditions form membrane-bound extracellular vesicles, known as EVs. The ability of these submicron structures to transfer their cargoes (miRNA, DNA, protein, cytokines, receptors, etc.) into recipient cells is described. Recent data revealed that platelet-derived extracellular vesicles (PEVs) crosstalk promotes cancer growth and metastasis formation. Moreover, they exert immunosuppressive activities on phagocytes. This EV subpopulation is the most abundant amongst all types in circulation. According to the authors' best knowledge, there is no information regarding the impact of PEVs on canine lymphocytes. The aim of this study was to evaluate the influence of PEVs on lymphocyte proliferation, phenotype and cytokine production in vitro. In the study, it was demonstrated (i) that PEVs interact differently with lymphocyte subsets and are preferentially associated with T-lymphocytes PBMC, while (ii) they are rarely detected in association with B-lymphocytes, and there is evidence that (iii) PEV uptake is observed after 7 h of co-culturing with lymphocytes. In addition, obtained data support the notion that PEVs do not influence in vitro lymphocyte proliferation, differentiation and cytokine production in a canine model.

The Influence of Intravaginal Gestagens Treatment on the Morphological Features and Endometrial Steroid Hormone Receptors Content during Anestrus Type II in Dairy Cattle.

BACKGROUND: Gestagens are the most widely used therapy in anestrus type II. The aim of this research is to evaluate the effectiveness of the vaginal progesterone inserts therapy in anestrus type II in cows. METHODS: The study was conducted on 33 cows. Progesterone (PR) and estrogen (ER) receptors expression in endometrium was assessed on a molecular level based on mRNA tissue expression. Additionally, blood 17ß-estradiol and progesterone levels were evaluated. RESULTS: A decrease in mRNA expression of A and B PR and ER α was noted in treated and untreated animals. In the treated group, an increase of ERß mRNA expression was observed, while a decreased was found in untreated animals. There was increased PR, ERα and ß expression in endometrial tissue in treated cows, and decreased expression of these factors in untreated cows. In the treated group, recurrence of ovarian cyclicity was noted in 52% of animals and pregnancy was obtained in 34.8% of them, while in the untreated group, recurrence did not occur. In the control group, spontaneous recurrence of ovarian cyclicity was not observed. An increase of PR expression was correlated with increased proliferation of endometrial cells. CONCLUSIONS: It seems likely that the endometrium is well developed and ready for placentation after removing the exogenous source of progesterone and preventing the recurrence of cyclicity of ovaries.

Thermal features, ambient temperature and hair coat lengths: Limitations of infrared imaging in pregnant primitive breed mares within a year.

Infrared thermography is a non-invasive technique which allows to distinguish between pregnant and non-pregnant animals. Detecting accurate body surface temperatures can be challenging due to external factors altering thermograph measurements. This study aimed to determine the associations between the ambient temperature, the hair coat features and the temperatures of mares' abdomens. It compared pregnant and non-pregnant mares throughout 11 months. The research was carried out on 40 Konik Polski mares, which were divided into pregnant and non-pregnant groups. The temperature (Tmax, maximal; Taver, average; Tmin, minimal) of the mares' abdomen was evaluated in two regions of interest: the whole area of the lateral surface of the mares' abdomen (Px1) and the flank area of the lateral surface of mares' abdomen (Px2). During the increasing period, the slopes in the linear regression equation did not differ significantly for ambient (Tamb) and surface temperatures in both groups. In the decreasing period, the slopes did not differ significantly for Tamb and Tmax in the non-pregnant group. They also did not differ for Tamb and Taver in Px1 and Tamb and Tmin in Px1 in both pregnant and non-pregnant groups respectively. Other slopes varied significantly (p < .001). There was no evidence of parallel changes in hair coat features and measured temperatures. The flank area appears more suitable for thermal imaging in pregnant mares due to the seasonal fluctuations in hair coat lengths.

The Physical Activity-Dependent Hematological and Biochemical Changes in School Horses in Comparison to Blood Profiles in Endurance and Race Horses.

Blood testing is one of the most important ways to improve performance, facilitate recovery and monitor the training of endurance and race horses. However, little is known about the physical activity-dependent changes of blood parameters in horses used for pleasure and in riding schools. This study aimed to perform routine blood tests for training monitoring of sport horses in three different horse types of use. Then the values of blood indicators were compared between school, endurance and race horses to find similarities in the physical activity-dependent profile. The study was carried out on 15 endurance, 15 race and 15 school healthy horses who underwent the typical effort for their disciplines. The hemogram parameters, creatine phosphokinase (CPK), aspartate aminotransferase (AST), blood lactate (LAC), and total serum protein (TSP) concentrations were measured using the same protocol and equipment. Measurements of main hematological and biochemical physical activity-dependent parameters were conducted before, immediately after and 30 min after training. In school horses, the physical activity-dependent increase of WBC (40.9%) and CPK (76.4%) was similar to endurance horses, whereas an increase of RBC (19.1%), HGB (18.6%) and HCT (19.4%) were more similar to race horses. The moderate effort-dependent increase of LAC concentration (2775%) was lower than in race horses (7526%) and higher than in endurance horses (390%). Limiting the training or work monitoring assessment of school horses to only the endurance or racing blood profile may result in the omission of significant changes in hematological and biochemical parameters.

The Effect of the Clenbuterol-ß2-Adrenergic Receptor Agonist on the Peripheral Blood Mononuclear Cells Proliferation, Phenotype, Functions, and Reactive Oxygen Species Production in Race Horses In Vitro.

Clenbuterol, the ß2-adrenoceptor agonist, is gaining growing popularity because of its effects on weight loss (i.e., chemical liposuction). It is also popular in bodybuilding and professional sports, due to its effects that are similar to anabolic steroids. However, it is prohibited by anti-doping control. On the other hand, it is suggested that clenbuterol can inhibit the inflammatory process. The cells from 14 untrained and 14 well-trained race horses were collected after acute exercise and cultured with clenbuterol. The expressions of CD4, CD8, FoxP3, CD14, MHCII, and CD5 in PBMC, and reactive oxygen species (ROS) production, as well as cell proliferation, were evaluated by flow cytometry. In addition, IL-1ß, IL-4, IL-6, IL-10, IL-17, INF-γ and TNF-α concentrations were evaluated by ELISA. ß2-adrenoceptor stimulation leads to enhanced anti-inflammatory properties in well-trained horses, as do low doses in untrained animals. In contrast, higher clenbuterol doses create a pro-inflammatory environment in inexperienced horses. In conclusion, ß2-adrenoceptor stimulation leads to a biphasic response. In addition, the immune cells are more sensitive to drug abuse in inexperienced individuals under physical training.