Identificazione di nuovi marcatori tumorali nelle neoplasie mammarie del cane: applicazione di tecnologie molecolari innovative

Canine mammary tumors (CMT) are the most common neoplasms in female dogs and the discovery of diagnostic, prognostic, and therapeutic markers is essential for improving the disease outcomes and the animal welfare. The aims of this study were to investigate the role of the Receptor tyrosine-kinase ErbB-2 (HER2) and to discover new potential tumor markers in CMT using innovative proteomic approaches. HER2 protein and RNA expressions were determined by immunohistochemistry (IHC) with an antibody extensively used in veterinary medicine and by real-time quantitative RT-PCR (qRT-PCR) as well as by Western Immunoblotting (WB) and Reverse-Phase Protein Arrays (RPPA). An orthogonal validation of HER2 protein expression was carried out using high-resolution mass spectrometry (MS). A diffuse cytoplasmic staining related to antibody lack of specificity was observed by IHC and further confirmed by WB. No differences between benign and malignant neoplasms were noted at mRNA level and the MS technique failed to detect HER2 peptides in the CMT. These results indicate a minor role of HER2 in CMT. Further, 40 proteins were found to be differentially expressed in normal mammary gland and in CMT by MS. Proteins related to glucose and to mitochondrial function as Transketolase, Transketolase like 1, and Prohibitin 2 were also investigate by IHC. The differences observed in hyperplasic and neoplastic lesions compared to normal mammary gland suggest these proteins as new potential markers for CMT

First evidence of intersex condition in extensively reared mullets from Sardinian lagoons (central–western Mediterranean, Italy)

The term intersex describes alterations in gonadal development with the simultaneous presence of male and female reproductive stages in the same gonad of a gonochoristic species. In coastal and estuarine environments, euryhaline fish living in polluted waters such as Mugilidae can frequently show these sexual anomalies. In this work, we analysed adult specimens of three species of euryhaline mullets (Chelon labrosus, Liza aurata and Mugil cephalus) from two Sardinian lagoons (Marceddì and San Teodoro) devoted to extensive aquacultural practices, in order to identify putative alterations in gonads and in gamete development. Overall, 13 of the 158 mullets examined (8.2%) were affected by gonadal disorders: four subjects (one C. labrosus, two L. aurata and one M. cephalus) exhibiting an intersex condition were found in the Marceddì lagoon and the other nine (five C. labrosus, two L. aurata and two M. cephalus) in the San Teodoro lagoon. Twelve of these gonads were classified as testis-ova (TOs) and one, belonging to a C. labrosus specimen, was a mixed gonadal tissue (MGT). Intersex condition was evaluated using an intersex index and all the recorded values showed a mild Ovotestis Severity Index (OSI). However, our findings suggest that fish gonadal disorders may be underestimated in extensive reared fish species, particularly in coastal brackish environments polluted by intensive agriculture and animal husbandry activities

Methylation dynamics during folliculogenesis and early embryo development in sheep

n/

"Architettura, scrittura, comunicazione, ricerca"

L’opera di architettura ci interroga riguardo a quale scrittura, quale comunicazione e quale lettura sia possibile dell’opera stessa. La scrittura richiama l’architettura così come l’architettura rimanda alla scrittura in quanto modelli su un foglio bianco

Absence of canine papillomavirus sequences in canine mammary tumours

Human papillomaviruses (PVs) are found in human breast cancer tissue; however, it remains controversial as to whether these viruses play a role in the aetiology of this tumour. There has been minimal study of whether PVs are found in normal or abnormal mammary glands of animals. The present study investigated whether a PV sequence could be found in the mammary glands of 33 female dogs by rolling circle amplification and polymerase chain reaction. No PV DNA was found in normal or neoplastic canine mammary tissues, suggesting that canine PVs are probably not involved in the pathogenesis of canine mammary neoplasia

Hypoluteoidism in a dog associated with recurrent mammary fibroadenoma stimulated by progestin therapy

Abstract Background Hypoluteoidism in the bitch is characterized by insufficient production and secretion of progesterone by the corpora lutea. It is a rare pathologic condition and during pregnancy, it leads to embryonic resorption or fetal abortion. Supplementary therapy with progestins is indicated during pregnancy to obtain delivery of vital puppies but unwarranted side effects of such treatment are poorly documented. Case presentation A 4-year-old, nulliparous, female Istrian Shorthaired Hound dog had been mated repeatedly in six heats with different dogs of proven fertility but signs of pregnancy did not develop. Estrous cycles, mating and pregnancies were monitored as hypoluteoidism or genital disease was suspected. During the first monitored estrus, the bitch was mated and on day 18 [day 0, day of estimated peak of luteinizing hormone (LH)], ultrasound examination showed three amniotic vesicles that were however found to be resorbed between day 20 and 23. Progesterone concentrations, measured by ELISA, were >8 ng/mL until day 12 and 1–2.5 ng/mL on days 20, 23 and 26. Primary hypoluteoidism was therefore suspected. In the second monitored estrus, the bitch was mated and during pregnancy, progesterone concentrations were >8 ng/mL until day 17 and 1–2.5 ng/mL on day 19. On days 20 and 22, two out of three embryonic vesicles had been resorbed. The bitch was treated with progesterone in oil from day 19 to day 58. Increase in the size of 2nd left thoracic mammary gland (T2-L) was observed and on day 46, ultrasound evaluation and biopsy were performed revealing a low-cellularity fibroadenoma. Parturition started spontaneously at day 65 but due to dystocia caused by fetal macrosomia, a Caesarean section was performed. During the next (third) monitored estrus, the bitch was bred again and during pregnancy, early decrease in progesterone concentration confirmed the diagnosis of primary hypoluteoidism. The bitch was treated with synthetic progestin (altrenogest) from day 8 to day 57. Five amniotic vesicles were detected by ultrasonography. Recurrence of swelling of T2-L was observed. On day 60, the bitch whelped five pups, two males and three females. As reported later by the owner, the latter did not show any sign of heat over the past 3 years. In one of them, clitoral hypertrophy and a blind ending vagina were diagnosed. Conclusions This is the first description of early hypoluteoidism in a pregnant bitch developing a mammary fibroadenoma under progestin treatment

Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization

Cryopreservation is routinely used to preserve cells and tissues; however, long time storage brings many inconveniences including the use of liquid nitrogen. Freeze-drying could enable higher shelf-life stability at ambient temperatures and facilitate transport and storage. Currently, the possibility to freeze-dry reproductive tissues maintaining vitality and functions is still under optimization. Here, we lyophilized sheep ovarian tissue with a novel device named Darya and a new vitrification and drying protocol and assessed effects on tissue integrity and gene expression. The evaluation was performed immediately after lyophilization (Lio), after rehydration (LR0h) or after two hours of in vitro culture (IVC; LR2h). The tissue survived lyophilization procedures and maintained its general structure, including intact follicles at different stages of development, however morphological and cytoplasmic modifications were noticed. Lyophilization, rehydration and further IVC increasingly affected RNA integrity and caused progressive morphological alterations. Nevertheless, analysis of a panel of eight genes showed tissue survival and reaction to the different procedures by regulation of specific gene expression. Results show that sheep ovarian tissue can tolerate the applied vitrification and drying protocol and constitute a valid basis for further improvements of the procedures, with the ultimate goal of optimizing tissue viability after rehydration

Capture myopathy in a corsican red deer <i>Cervus elaphus corsicanus</i> (Ungulata: Cervidae)

The present study focused on the importance of translocation as stressful event in an endangered cervid species causing fatal consequences during capture operations. Fourteen free-ranging Sardinian red deer (Cervus elaphus corsicanus) have been captured for restocking propose in a protected area of southwestern Sardinia. The cervids were chemically immobilized, transported to a restocking area and released in the wild. Sixteen hours after the release, a pregnant hind was found dead and a complete necropsy was performed. Post-capture blood samples showed increased levels of creatine kinase, lactate dehydrogenase, cortisol and potassium and were highly indicative of stress-linked muscle damage. The macro- and microscopic lesions consisted of muscular and cardiac degeneration, and renal injury. In the kidneys, the presence of myoglobin in intratubular casts, detected by immunohistochemistry assay, suggested an acute renal failure associated with myoglobinuric nephrosis as a consequence of rhabdomyolysis. The pathological findings were indicative of capture myopathy in a Cervus elaphus corsicanus. This condition has never been reported in the endangered Corsican red deer. This report underlines that mortality from capture is a risk that must be considered during restocking programs. Further studies are needed to minimize stress to preserve wildlife from the risk of fatal consequences due to human interactions