Kisspeptin and bovine follicular cysts.

The aim of this study was to investigate kisspeptin (Kp) and sexual hormone blood concentrations in healthy dairy cows (H) and in cows diagnosed with ovarian follicular cysts (FC). Forty dairy cows were enrolled in this study and divided in 2 groups of 20 subjects each. All cows underwent blood collection on the day of diagnosis and on that of heat detection, respectively. Kisspeptin 10, estradiol, and progesterone blood concentrations were assessed. All the parameters were higher in Group FC than in Group H. These results suggest that the higher steroid levels found may have determined the increase in Kp secretion in the cystic cows. This, in turn, may have directly or indirectly stimulated the sustained Luteinizing Hormone (LH) release, known to occur in dairy cystic disease.

In vivo model for mastocytosis: A comparative review.

Human mastocytosis are heterogeneous group of neoplastic diseases characterized by a different degree of uncontrolled mast cell (MC) proliferation and activation. Interestingly, human mastocytosis share several biological and clinical features with canine mast cell disorders, so called canine mast cell tumors (CMCTs). These CMCTs are the most common spontaneous cutaneous tumors found in dogs representing a valid model to study neoplastic mast cell disorders. It has been discovered that the pathological activation of c-Kit receptor (c-KitR), expressed by MCs, has been involved in the pathogenesis of neoplastic MC disorders. In this review we have focused on human mastocytosis in terms of: (i) epidemiology and classification; (ii) pathogenesis at molecular levels; (iii) clinical presentation. In addition, we have summarized animal models useful to study neoplastic MC disorders including CMCTs and murine transgenic models. Finally, we have revised therapeutic approaches mostly common in human and canine MCTs and novel tyrosine kinase inhibitors approved for CMCTs and recently translated in human clinical trials.

Immunological aspects of metritis in dairy cows: a review.

This paper reviews puerperal metritis in the cow, particularly the complex and multi-factorial pathogenesis characterized by an altered cross-talk among infectious agents, endocrine and immune systems. Uterine infections impair fertility and is one of the main causes of economic losses in dairy production. The early postpartum is a period characterized by an increased exposition to infectious agents and the disruption of the metabolic homeostasis, leading to endocrine and immunologic disorders. Dysregulation of uterine defence mechanisms results in the development of metritis. Because there is a complex interaction between infectious, endocrine and immune factors during metritis, there is need to use safer and cheaper drugs which are able to strengthen the anti-infective actions of the routine therapies.

Effects of mefepronic acid (2-phenoxy-2-methyl propionic acid) on hepatic metabolism and reproductive parameters in postpartum dairy cows.

This study investigates the effects of mefepronic acid (MA), a PPAR-α agonist, on hepatic metabolic functions and reproduction of postpartum dairy cows. Sixty Friesian cows were divided into Group A (administered 5g of MA IM, within 24 hrs after calving, on the 3(rd) and 5(th) day postpartum) and Group B (control). All the cows were blood sampled within 24 hrs of calving (Day 0), on Day 3, 5, 10, 15, 30, and 40 postpartum. On plasma, metabolic and biochemical parameters were determined. Liver biopsies were performed on Day 0, 15 and 30 for the evaluation of hepatic lipid and glycogen content. Reproductive parameters were also evaluated. In Group A, blood HDL, glucose and cholesterol increased till the end of the study, in accordance with the histological results. PPAR-α immunopositive cells increased in liver slices of Group A, too. Reproductive parameters improved in Group A. This study highlights the beneficial effects of mefepronic acid on the hepatic metabolism and reproductive parameters of post-partum dairy cows.

Tyrosine kinase inhibitors (TKIs) in human and pet tumours with special reference to breast cancer: a comparative review.

Tyrosine kinase receptors (TKRs) play a key role in tumour cell proliferation and survival since they are involved in endothelial cell activation leading to tumour neoangiogenesis. In particular, vascular endothelial growth factor receptors (VEGFRs), platelet-derived growth factor receptor (PDGFR), stem cell factor receptor (c-KitR), and colony-stimulating factor 1 (CSF-1) are overexpressed or constitutively activated in human and pet malignancies. A variety of small molecule inhibitors targeting specific tyrosine kinases (known as tyrosine kinase inhibitors or TKIs) have recently been approved, or are under investigation, for the treatment of human cancer. TKI application in animal cancer is however relatively recent. This review aims to illustrate the major aspects of tyrosine kinase dysfunctions, with special regard to human and animal cancer of the mammary gland, providing an update on the background of the anti-angiogenic and anti-neoplastic properties of TKIs in human and veterinary cancer.

In vitro effects of L-arginine on spontaneous and homocysteine-induced contractility of pregnant canine uteri.

The L-Arginine-Nitric Oxide Synthase-Nitric Oxide (L-Arg-NOS-NO) system exerts a pivotal role in the maintenance of uterine quiescence during pregnancy, whereas Homocysteine (Hcy) promotes uterine contractility. The aim of this study was to test the in vitro effects of L-Arg on spontaneous and Hcy-induced contractions of uteri excised from pregnant bitches. 104 strips cut from pregnant uteri were mounted in an organ bath. 40 out of 104 strips (16 from mid-gestation uteri and 24 from close to term uteri, respectively) were exposed to cumulative doses of L-Arg; 40 strips (16 from mid-gestation-uteri and 24 from close to term-uteri, respectively) were exposed to N-nitro-L-arginine methyl ester (L-NAME), a NOS antagonist; the remaining 24 strips (from close-to-term uteri) were first exposed to a single dose of Hcy and then to increasing doses of L-Arg. L-Arg showed no effects on spontaneous contractility both in mid-gestation- and close to term-uterine strips, whereas it promoted a relaxant effect on Hcy-induced contractility. On the contrary, L-NAME increased amplitude of contraction both in mid-gestation and close to term strips. These findings suggest that the L-Arg-NO system is present in the uterus of pregnant bitches and that Hcy is able to modulate its actions. Further investigation of this system may provide the basis of future obstetrical therapies in bitches.

Epidural vs intramuscular administration of lecirelin, a GnRH analogue, for the resolution of follicular cysts in dairy cows.

Bovine follicular cysts are an ovarian disorder of dairy cows associated with abnormal estrous behaviour and infertility. The treatment of choice is intramuscular administration of a GnRH analogue, which acts by triggering pituitary release of LH. However, the presence of GnRH and GnRH receptors on spinal cord and ovary in some species, and the kind of innervation of the ovary, let us hypothesize that GnRH and its analogues may also act when administered by epidural route, as happens for other drugs. Therefore the aim of this study was to compare the effects of epidural vs intramuscular administration of lecirelin (a GnRH analogue) on FC regression, estrus detection and pregnancy outcomes. The study was conducted on 220 Friesian cows affected by follicular cysts, divided among 4 groups: Group L(epid) and Group L(im) received, respectively 50 µg of lecirelin in the epidural space and intramuscular; Group C(epid) and Group C(im) were used as control groups. In Group L(epid), estrus induction and pregnancy rates were significantly higher than in Group L(im). The results of this study show that the epidural administration of lecirelin promoted the remission of follicular cysts and an improvement of reproductive parameters compared to intramuscular administration. Thus, an alternative therapeutical approach is available for FC treatment, in order to obtain an easier restoration of the ovarian activity, especially in those cases refractory to classical therapeutic approaches.

In vitro activity of human chorionic gonadotropin (hCG) on myometrium contractility.

LH/hCG receptor has been found in extragonadal tissues in human and animals. The myometrium presents such receptors but their functional role is still not clear. Aim of our study was to test the activity of human chorionic gonadotropin (hCG) on bovine uterine contractility. Uterine strips from cows both during follicular and luteal phases were mounted in an organ bath and then exposed to increased doses of hCG. The amplitude of the myometrium contractions were significantly decreased in the follicular and luteal phase but the frequency was not affected. These findings prove a relaxing effect of hCG in the bovine uterus, as already shown in the sow and human, and its possible functional role in modulating uterine contractility.

Uterine contractility is strongly influenced by steroids and glucose metabolism: an in vitro study on bovine myometrium.

BACKGROUND: Energetic substrates and hormonal phase are important for uterine contractions. Etomoxir a muscle carnitine palmitoyltransferase-1 inhibitor, able to diverge uterine metabolic pathways towards glycolysis, facilitates glucose utilisation. The aim of this study was to evaluate its effect on uterine contractility in different hormonal situations. METHODS: Uterine samples were collected from 60 cows during follicular phase, luteal phase and pregnancy. The cows were slaughtered at a local abattoir. Longitudinal strips were mounted vertically in a 30-ml organ bath connected to an isometric force transducer. Contractions were recorded with an ink-writing polygraph. After the equilibration period, etomoxir was added to the organ bath at different concentrations. The amplitude and frequency of contractions were registered before and after addition of etomoxir. RESULTS: In 17 strips from pregnant cows, etomoxir increased the amplitude (p < 0.05) of contractions but not the frequency in comparison with basal conditions. In 15 strips from cows in the luteal phase, etomoxir increased the amplitude (p < 0.05) and frequency of contractions (p < 0.05 at 5 µM and p < 0.01 at 8 and 10 µM). In 18 strips from cows in the follicular phase, etomoxir increased the frequency of contractions but not the amplitude (p < 0.01 at 5 µM and p < 0.05 at 8 and 10 µM). CONCLUSIONS: This study confirms the importance of glucose for uterine contractility and, moreover, it underlines different patterns of contraction with regard to the hormonal status.

Bovine ovarian follicular cysts: in vitro effects of lecirelin, a GnRH analogue.

This study investigates the mechanisms of action by which a GnRH analogue may modulate the contractility of the bovine ovarian follicular wall. The in vitro evaluation of the spontaneous basal contractility of bovine preovulatory and cystic follicles was performed, followed by testing the effects of lecirelin, a GnRH analogue, on their basal contractility. Strips of tissue in isolated organ bath were employed. In addition, to better investigate the mechanism of action of lecirelin, the study of the effects of cumulative doses of nifedipine (a calcium channel blocker), phentolamine (an α-adrenoceptor antagonist) and reserpine (an inhibitor of the vesicular up-take of catecholamines) alone and, at the highest doses employed, associated to lecirelin, was set up. The results demonstrate that in basal conditions and after the addition of lecirelin, the strips from preovulatory follicles contract significantly more than strips from cysts. Furthermore, among the patterns of contractility evoked by the three drugs employed, the one induced by nifedipine was the only one unaffected by the addition of lecirelin. The data obtained provide the hypothesis that one of the main mechanisms of action of GnRH, could involve calcium channels.

Peritoneal adhesions in human and veterinary medicine: from pathogenesis to therapy. A review.

Any peritoneal inflammatory process consequent to infections or surgical injuries may induce abdominal adhesion formation. Peritoneal adhesions are connective laciniae that develop among abdomino-pelvic organs that limit physiologic visceral motion. Consequently, fertility may be impaired, and intestinal obstruction and pelvic pain may develop, mainly in subjects that had undergone gynaecological surgery. This review illustrates the pathogenic steps of adhesiogenesis and the therapeutic scenario that evolved over the years to tackle the threat of peritoneal adhesions, both in domestic animals and in women.

Follicular development in pregnant cows after the administration of equine chorionic gonadotropin (eCG): a new insight.

The follicular development in the cow occurs in a wave-like pattern, and it takes place also during pregnancy. In the cow, Equine Chorionic Gonadotropin (eCG) is used for superovulation, but a decrease in total fertility has been reported, likely because of its immunogenic properties in species other than equine. In this regard, immune response has been implicated in follicular growth, ovulation, and placental development. So, aims of our study are to test the safety of eCG administered during pregnancy and characterize the ovarian activity, the quality of oocytes, the hormonal status, and interleukin levels in eCG-treated pregnant cows.

Reactive oxygen species (ROS): involvement in bovine follicular cysts etiopathogenesis.

Ovulation is compared to an acute inflammatory process during which vasoactive agents, prostanoids, leukotrienes and Reactive Oxygen Species (ROS) develop. The aim of this study was to evaluate the levels of ROS in cystic and follicular fluid, in order to establish their involvement in the etiopathogenesis of Cystic Ovarian Follicle (COF) in dairy cows. The study was conducted in 30 healthy cows (group C) and 30 cows affected by COF (group COF). The fluid of follicular cysts and of preovulatory follicles was drawn by means of ultrasound guided aspiration from the cows of both groups. The fluid obtained was analyzed by a photometric analytical system to detect ROS level. ROS concentration was statistically lower in the cystic fluid than in the follicular one (62.4 +/- 13.36 U.Carr vs. 84.89 +/- 26.99 U.Carr) (p<0.05), thus suggesting that an alteration of the cascade responsible for ROS production may be implicated in the complex etipathogenesis of COF.

Effects of antibiotics on biochemical parameters, leukocytes and reactive oxygen species (ROS) in bitches after ovariectomy.

It is well known that following surgical procedures, a variety of patho-physiological alterations occurs in the host, such as changes in haemodynamic, endocrine and immune functions, as well as increase in Reactive Oxygen Species (ROS). Antibiotic administration, a common practice used in surgery, affects immune functions and ROS generation. Our study was aimed to investigate the effect of five different antibiotic regimens: amoxicillin, benzylpenicillin/dihydrostreptomycin, sulfametazine/sulfamerazine/ sulfathiazole, enrofloxacin, lincomycin/spectinomycin, administered twice (2 hours before skin incision and 6 hours after the end of the surgical suture), on biochemical parameters, leukocytes and ROS concentrations, in bitches undergoing open laparotomic ovariectomy. All treated bitches recovered from surgery without developing either systemic dysfunctions, or infections at the surgical site. Mean healing time was 7 +/- 3 days. An increase in white blood cell count, in differential leukocyte count and in ROS concentrations occurred 24 hours after the end of surgery, followed by a gradual decrease to basal values within the fourth day after surgery. The biochemical parameters remained quite constant throughout the study. In conclusion, our results show that the above antibiotic regimens, administered only twice, are efficient in inducing an uneventful recovery after laparotomic ovariectomy and preventing the development of infections in the bitch.

Serum levels of reactive oxygen species (ROS) in the bitch.

The aim of this study was to determine the serum concentrations of reactive oxygen species (ROS) during the different phases of the estrous cycle in the bitch, in order to establish their physiological values. 56 healthy mixed-breed bitches were enrolled at this purpose and divided into 4 groups, standing on the different phases of the estrus cycle. Blood samples were collected in all groups and serum ROS concentrations were determined. Proestral concentrations were statistically higher than anestral ones, and statistically lower than those found in estrus (p<0.001). The highest concentrations of ROS were detected at estrus, that is, in the peri-ovulatory period. This sharp increase in ROS concentrations is related to the acute inflammatory process underlying ovulation and to the increase in immune and metabolic activities, cytological changes and myometrial contractility promoted by the high levels of estrogens. In diestrus, the mean concentration of ROS decreases. This reduction did not show any statistically significant difference with the mean value observed in proestrus. In this phase, in fact, the high concentrations of progesterone, exerting an antioxidant and immunodepressive effect, justify the lower mean concentration of ROS detected. In anestrus, the lowest concentrations of ROS were observed, for the reduced metabolic and endocrine activity occurring in this phase of the estrous cycle. In conclusion our results establish the physiologic levels of ROS during the estrous cycle in the bitch and reflect the endocrine morphologic and metabolic changes occurring during it.

Thyreotropic effect of human Chorionic Gonadotropin (hCG) in mares at estrus.

The beta-subunits of thyroid stimulating hormone (TSH) and human chorionic gonadotropin (hCG) share a high homology, just like the ectodomains of their receptors, do. As a consequence, hCG was shown to exert a thyrotropic action in humans and hamsters. This study aimed to investigate whether hCG, used to induce ovulation, displays a thyrotropic effect in the equine species too. Forty mares at estrus were divided in two groups; 20 were intravenously treated with sterile saline solution (NaCl 0.9%) (group A); 20 were intravenously treated with 4000 I.U. of hCG (group B). All the mares were artificially inseminated 24 hours after the administration of the drugs and underwent blood collection at estrus detection (T1), 2, 6, 24 hours later (T2, T3, T4), and 3 and 6 days later (T5, T6), in order to detect serum fT3 and fT4 concentrations. fT3 concentrations gave rise to similar and constant trends in the two groups, without any statistically significant difference, whereas fT4 concentrations declined in both groups, remaining statistically higher in group B than in group A for 48 hours (p < 0.05), thus accounting for a thyrotropic effect of hCG. Pregnancy rate was 70% in group B and 40% in group A, respectively. The higher success in the outcome of pregnancy may result from the immunomodulating and steroidogenic effects of hCG and from the higher levels of thyroid hormones observed in the hCG treated group.

First demonstration of an increased serum level of reactive oxygen species during the peripartal period in the ewes.

Reactive Oxygen Species (ROS) are produced during oxidative metabolism, and regulate many biological processes. The acute inflammation characterizing parturition induces many physiological changes. Among them, there is evidence that ROS affect the synthesis of many factors involved in parturition. Our study aims to determine serum levels of ROS in periparturient ewes, as well as to establish a value of reference of their physiological concentration. ROS determination was performed on blood collected every 12 hours in periparturient twin pregnant ewes. Our results will show a significant increase in ROS concentrations from the beginning to the end of the experiment. This increase may be due to the inflammatory process establishing during parturition.