Marianna Pantaleo

Oxidative Stress in Neonatology. A Review

Free radicals are highly reactive oxidizing agents containing one or more unpaired electrons. Both in human and veterinary neonathology, it is generally accepted that oxidative stress functions as an important catalysator of neonatal disease. Soon after birth, many sudden physiological and environmental conditions make the newborn vulnerable for the negative effects of oxidative stress, which potentially can impair neonatal vitality. As a clinician, it is important to have in depth knowledge about factors affecting maternal/neonatal oxidative status and the cascades of events that enrol when the neonate is subjected to oxidative stress. This report aims at providing clinicians with an up-to-date review about oxidative stress in neonates across animal species. It will be emphasized which handlings and treatments that are applied during neonatal care or resuscitation can actually impose oxidative stress upon the neonate. Views and opinions about maternal and/or neonatal antioxydative therapy will be shared.

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.

Localization of thyrotropin receptor and thyroglobulin in the bovine corpus luteum

The receptor of the Thyroid Stimulating Hormone (TSHr) and thyroglobin (TGB), are two proteic factors necessary for the synthesis of hormones, in the thyrocite. In mammals, many immuno-histochemical reports indicate the presence of the TSHr in extra-thyroidal tissues, but not in the ovary. Triiodothyronine (T(3)) and thyroxine (T(4)) have been widely shown to affect ovarian functions and the synthesis of progesterone (P(4)). The aim of this study was to determine if by immunohistochemistry techniques TSHr and TGB could be found in the bovine corpora haemorragica, lutea and albicantia. A primary rabbit polyclonal antibody against human TSHr and a primary rabbit polyclonal antibody against human TGB were employed. Furthermore, the accuracy of bovine thyroid to the antibodies used in this study was tested. A positivity reaction for the anti-TSHr serum in the large luteal cells and immunostaining of both small and large luteal cells with the anti-TGB serum occurred only in mature corpora lutea. No immunostaining was detected in stromal cells, blood and lymphatic vessels and in corpora haemorragica and albicantia. Bovine thyroid tissue showed immunostaining to both the antibodies employed. These data suggest that the luteal cells of mature corpora lutea may be involved in the synthesis of thyroid hormones, which may modulate P(4) synthesis, acting in an autocrine and paracrine way.

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.

Evaluation of blood and milk oxidative status during early postpartum of dairy cows

In dairy cows, the intensity of metabolic activity, associated with the negative energy balance (NEBAL), is responsible for an increased production of reactive oxygen species (ROS) and, subsequently, for the development of the condition of oxidative stress, which may overwhelm the antioxidant potential of the bovine maternal organism, making it prone to the development of many puerperal dysfunctions, as well as to an alteration of colostrum and milk quality. Given these premises, the aims of this study are to evaluate serum and milk concentrations of ROS and lipoperoxides, vitamins A and E, on the 10th, 12th, 14th and 16th day postpartum of dairy cows, a particularly critical period during which the NEBAL reaches its nadir, and to compare the trends of these parameters in two different bovine breeds. The study was performed in pluriparous Italian Friesian and Brown dairy cows. On the 10th day postpartum, all cows underwent a clinical examination to exclude the presence of alterations; furthermore, on the same day, a milk sample was collected from each cow, in order to perform the somatic cell count (SCC; (CE) N. 853/2004) and to establish which of them had an SCC ≤400,000/ml or >400,000/ml. In this study, among the 110 cows that were initially selected, the evaluation of these parameters allowed the inclusion of 80 animals, which were divided into four groups of 20 subjects each: Group F and F1: Italian Friesian healthy cows, with SCC ≤400,000/ml and >400,000/ml, respectively; Group B and B1: Italian Brown healthy cows, with SCC ≤400,000/ml and >400,000/ml, respectively. On the 10th, 12th, 14th and 16th day postpartum, peripheral blood and milk samples were collected. The results obtained show that in group B1 there were higher concentrations of ROS and milk antioxidants compared with Friesian group cows. This datum let us suppose that even in the presence of higher ROS concentrations the antioxidant status found in group B1 seems to be able to counteract the oxidative damage, which is more likely to develop in these cows.

Blood and milk oxidative status after administration of different antioxidants during early postpartum in dairy cows.

This paper describes serum and milk oxidative profiles during early postpartum (10th-16th day) of dairy cows, after three different treatments: β-carotene and α-tocopherol (Dalmavital®, Fatro, Italy) (VAE Group); L-carnitine, d, L-acetylmethionine, cyanocobalamin (Adiuxan®, Ceva Vetem, Italy) (LCAR Group) and Dalmavital®+Adiuxan® (VAE/LCAR Group). Antioxidant administrations resulted in a general reduction in ROS concentrations which decreased more after a single drug administration (VAE and LCAR Groups) vitamin A and E serum levels increased in the three treated groups compared to control one (p < 0.05). Milk lipoperoxide content was statistically lower in treated cows than in control ones, whereas, as to vitamins, only vitamin E increased in all the treated groups compared to control. Taken together, the results of this study confirm the efficacy of antioxidant administration in the early postpartum of dairy cows and suggest that an excess in antioxidant administration may result, paradoxically, in an increase in ROS generation.

In vivo and in vitro studies of the role of the adrenergic system and follicular wall contractility in the pathogenesis and resolution of bovine follicular cysts.

Bovine follicular cysts (FCs) are a common cause of economic loss in modern dairy herds. Their aetiopathogenesis is not completely understood, even though an inadequate hypothalamic release of GnRH at the time of ovulation is considered to be their main cause. Much evidence, however, suggests a role for adrenergic innervation in ovarian functions, such as follicular development, steroid hormone secretion, and follicular contractility, the latter being an event important for ovulation. Moreover, in humans, polycystic ovary syndrome, a disease very similar to bovine follicular cysts, is characterised by increased density of adrenergic nerves. Given these premises, the aim of our study was to evaluate the effectiveness and mode of action of a novel strategy for the treatment of bovine follicular cysts. In the in vivo experiment, 170 Friesian cows diagnosed with follicular cysts were assigned to four groups (groups A, B, C, and D) to assess the effects of epidural administration of a β-adrenergic antagonist (carazolol) alone or in combination with a GnRH analogue (lecirelin). The four groups underwent the following treatments: Group A was administered lecirelin in combination with carazolol; Group B was administered carazolol; Group C was administered lecirelin; and Group D was administered only normal saline solution. In the in vitro experiment, strips of the walls of cystic follicles recovered post-mortem were suspended in an organ bath, connected to an isometric force transducer and exposed to increasing doses of epinephrine or to the same treatment after exposure to carazolol for 15 min (n = 10). The amplitude and frequency of the contractile activity were recorded. None of the control cows was observed in oestrus or was submitted to AI. The combination of lecirelin and carazolol induced a significant increase in the number of cows in oestrus (88%) compared to lecirelin alone or to carazolol alone (P < 0.05 and P < 0.01, respectively). The combination of lecirelin and carazolol and lecirelin alone were significantly more efficacious than carazolol alone (P < 0.01 and P < 0.05, respectively). In the in vitro experiment, epinephrine increased the amplitude of the contractions of the strips in a dose-dependent manner. This response was significantly enhanced in strips pre-treated with carazolol. The treatments had no effect on the frequency of contractions. In conclusion, our work demonstrates that the epidural administration of a GnRH analogue and a β-adrenergic blocker is an effective means of treating cows with cystic ovarian disease. Moreover, it confirms, from a clinical point of view, that alterations of the adrenergic system and of the contractility of the follicular wall can be considered aetiopathogenic factors involved in the development of FCs. The results of this study lay the basis for a new therapeutic approach to FCs.

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.

Evaluation of serum concentrations of interleukin (IL)-4, IL-10, and IL-12 during pregnancy in bitches

Complex cytokine networks play an important role in a wide range of pregnancy-related processes. During physiological pregnancy, the balance of T helper lymphocytes Th1 and Th2 is strongly shifted toward Th2, which has a protective role in the feto-maternal interaction. The aim of the present study is to establish the serum concentrations, ranges, and trends of anti-inflammatory interleukin (IL)-4 and IL-10, and inflammatory IL-12A, during three phases of gestation in the bitch (20-30, 31-40, and 41-57 days of gestation). Our results indicate that, in early gestation, IL-4 and -10 serum concentrations are elevated and might depend on progesterone that could act as a potent inhibitor of Th1 responses inducing, conversely, the production of Th2-type cytokines (i.e., IL-10 and IL-4). On the other hand, between 30 and 40 days of gestation, the concentrations of the anti-inflammatory ILs decrease probably because of high concentrations of prolactin, which is endowed with immunostimulatory properties on different immune cell types. In the third phase of gestation (41-57 days), an increase in IL-10 occurs, which might depend on high levels of 17β-estradiol that, during pregnancy, interfere with the ability of dendritic cells to stimulate T lymphocytes, acting as an anti-inflammatory factor. Conversely, the low and persistent concentrations of inflammatory IL-12A, throughout pregnancy, compared with anti-inflammatory ILs, might depend on the characteristic cytokine products of Thl and Th2 cells that are known to be mutually inhibitory. The monitoring of immunological status via the levels of cytokines during pregnancy in the bitch, could represent a diagnostic tool to predict and/or prevent pregnancy abnormality, as demonstrated in women.

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

Abstract 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.