Investigation of efficacy of two different chemotherapy protocols used in neoadjuvant chemotherapy in clinical stages II-IV canine malignant mammary tumours.

The first aim of this study is to demonstrate the clinical efficacy and reliability of two different neoadjuvant chemotherapy (NAC) protocols consisting of doxorubicin/cyclophosphamide (AC) and paclitaxel in dogs with clinical stages II-IV canine malignant mammary tumours (CMTs). Secondly, to determine the Luminal A, Luminal B, HER2-positive and triple-negative molecular subtypes and their value in predicting clinical response to NAC in biopsy samples, and thirdly, to reveal the changes in Ki-67, human epidermal growth factor receptor type 2 (HER2), oestrogen receptor (ER), and progesterone receptor (PgR) expression levels induced by NAC. Thirty dogs with clinical stages II-IV CMTs (T1-3N0-1M0) according to the modified TNM system were included in the study. Dogs in group-1 (n = 15) AC combination and dogs in group-2 (n = 15) were administered paclitaxel. Partial response (PR) was the most common clinical response in both treatment groups (66.66% and 86.66%, respectively). There was no difference between the groups regarding clinical response parameters (p = .001). The rate of treatment responders was higher than the rate of non-responders in both groups (p < .001). The adverse effects observed in both groups were mostly limited to grades 1 and 2 and all were easy to manage. The most frequently detected molecular subtype was Luminal A (59.25%). Complete response (CR) was achieved in 33.33% of dogs with triple-negative CMT in the AC group and 14.29% of the Luminal A subtype in the paclitaxel group. Alterations in Ki-67, HER2, ER, and PgR expressions after chemotherapy were not statistically significant (p > .05). As a result, we have shown that these neoadjuvant chemotherapy protocols are effective and safe alternative treatment options for CMTs.

Anti-Müllerian hormone: A novel biomarker for detecting bovine freemartinism.

The anti-Müllerian hormone (AMH) indicates ovarian reserve in cattle, maintaining a consistent trajectory post-puberty. In heterosexual pregnancies, the development of the Müllerian duct in female foetuses is inhibited, resulting in an anticipated minimal or absent ovarian reserve capacity. This investigation aimed to compare AMH levels in healthy Holstein heifers that had reached puberty with those of freemartin animals of the same breed and age. The study incorporated Holstein heifers reaching puberty between 11 and 15 months of age in Group 1 (G1, n = 20) and freemartin animals in Group 2 (G2, n = 19, 16). AMH measurements (AMH-1/AMH-2) were recorded at 12-day intervals for the study participants. Notably, AMH levels in three freemartin animals could not be detected, prompting statistical analysis based on measurements from the remaining 16 freemartin animals in G2. A statistically significant correlation was observed between two separate measurements in G1 and G2 (p < .001). Furthermore, AMH-1 and AMH-2 levels were statistically higher in G1 than in G2 (p < .001). In G1, AMH-1 levels ranged from 227 to 677 pg/mL, with an average of 367.3 ± 25.5 pg/mL, and AMH-2 levels ranged from 234 to 645 pg/mL, with an average of 380.8 ± 24.4 pg/mL. Conversely, in G2, AMH-1 levels ranged from 10 to 72 pg/mL, with an average of 26.8 ± 4.44 pg/mL, and AMH-2 levels ranged from 12 to 68 pg/mL, with an average of 28.75 ± 4.18 pg/mL. The mean AMH levels in G1 were approximately 14 times higher than in G2 (p < .001). Consequently, ROC analysis utilizing AMH-1 and AMH-2 data established cut-off values of ≤72 and ≤ 68 pg/mL respectively for distinguishing freemartin animals. In conclusion, AMH could be used as a reliable biomarker for identifying Holstein freemartin animals.

Evaluation of serum anti-Müllerian hormone concentrations following treatment with vitamin D in Holstein Friesian heifers.

Anti-Müllerian hormone (AMH) levels in the blood of dairy cattle are associated with the count of antral follicles (AFC), ovarian function. Numerous studies, particularly in human medicine, have shown that vitamin D3 (Vit D3) has a positive effect on AMH levels. To our knowledge, the effects of Vit D3 on serum AMH levels in heifers have not been investigated. The goal of the current study was to evaluate the concentrations of serum AMH in dairy heifers following Vit D3 treatment. The study included 20 healthy non-pregnant Holstein Friesian heifers. These animals came to estrus at least once. All heifers received a single intramuscular dosage of 5 million IU Vit D3 (Cholecalciferol). Blood samples were collected from the coccygeal veins of all animals before and after Vit D3 injection (days 7, 14, and 28) for the measurement of AMH and 25(OH)D concentrations. In our analysis, we observed that after exogenous Vit D3 administration, circular 25(OH)D increased constantly (up to day 28 after Vit D3 injection) in all animals. On day 28, AMH concentrations saw a 10% increase in comparison with those measured the day before the Vit D3 injection. However, a comparison of AMH concentrations measured across days did not reveal statistically significant differences between Day 0, 7, 14, and 28 levels (p = .10). Furthermore, no statistical correlation was observed between the pairs (age-AMH) and (weight-AMH). Considering all times, no correlation was found between 25(OH)D and blood AMH levels. These findings demonstrated that exogenous Vit D3 did not affect serum AMH in Holstein Friesian heifers.

Fertility of Holstein heifers after two doses of PGF2α in 5-day CO-Synch progesterone-based synchronization protocol.

The objective of the study was to determine the effect of three different PGF2α (PGF) treatments in the 5-day CO-Synch progesterone-based synchronization protocol on artificial insemination (AI) pregnancy rate (PR) in Holstein heifers in Turkey and the United States. We hypothesized that two doses of PGF administered concurrently or 6 hours apart would result in greater AI pregnancy compared with a single dose of PGF on Day 5 at controlled internal drug release (CIDR) removal. In Turkey, Holstein heifers (n = 450) from one farm in the province of Adana and another farm in the province of Bursa were included. In the US, Holstein heifers (n = 483) from two locations in the state of Idaho were included. Heifers within locations were randomly allocated to one of three protocol groups: 1PGF-received 25 mg IM of dinoprost at CIDR removal; 2Co-PGF-received 50 mg IM of dinoprost at CIDR removal, and 2PGF-received 25 mg IM of dinoprost at CIDR removal and an additional 25 mg IM of dinoprost 6 hours later. All heifers received a CIDR (1.38 g of progesterone) and GnRH (10 µg IM of Buserelin [Turkey] or gonadorelin hydrochloride [US]) on Day 0. The CIDRs were removed on Day 5, and each heifer was given PGF according to the assigned treatments. On Day 7, each heifer was given another dose of GnRH and concurrently inseminated at 56 hours after CIDR removal. Heifers in both experiments were examined for pregnancy status between 35 and 45 days after AI. Overall, controlling for age, the heifers in the 2PGF group had greater AI-PR (61.7% [192/311]) than heifers in 2Co-PGF (48.2% [149/309]; P < 0.001) or 1PGF (53.7% [168/313]; P < 0.05) groups. No difference was observed between 2Co-PGF and 1PGF groups (P > 0.1). In Turkey, the heifers in the 2PGF group had a greater AI-PR (60% [90/150]) than 2Co-PGF (45.3% (68/150); P < 0.01] group. No difference was observed between 2PGF and 1PGF (55.3% [83/150]) groups (P > 0.1). There was a trend for AI pregnancy between 1PGF and 2Co-PGF groups (P = 0.08). In the United States, the heifers in the 2PGF group had a greater AI-PR (63.4% [102/161]) than the heifers in 2PGF (50.9 [81/159]; P < 0.05) or 1PGF (52.1% [85/163]; P < 0.05) groups. Heifers that were 15- and 16-month old achieved greater AI-PR than 17- and 18-month-old heifers (59.2 [342/578] vs. 47.0% [168/355]; P < 0.01). In conclusion, administration of 2PGF at 6 hours apart on Day 5 at CIDR removal in a 5-day CO-Synch + CIDR protocol resulted in greater AI pregnancy. A greater number of 15- and 16-month-old heifers became pregnant compared with 17- and 18-month-old heifers.

Using cell banks as a tool in conservation programmes of native domestic breeds: the production of the first cloned Anatolian Grey cattle.

The aim of this study was to clone native Anatolian Grey cattle by using different donor cell types, such as fibroblast, cartilage and granulosa cells cryopreserved in a gene bank and oocytes aspirated from ovaries of Holstein cows as the recipient cytoplasm source. One male calf from fibroblast, three female calves from granulosa cells and one female calf from cartilage cells were born healthy and at normal birthweights. No calves were lost after birth. The results demonstrated that the cloned calves had the same microsatellite alleles at 11 loci as their nuclear donors. However, the mtDNAs of the five Anatolian Grey cloned calves had different haplotypes from their donor cells and mtDNA heteroplasmy could not be detected in any of the clones. The birth of healthy clones suggests that the haplotype difference between the cell and oocyte donor did not affect the pre- or post-implantation development of the bovine nuclear transfer derived embryos in our study. The results showed that well established nuclear transfer protocols could be useful in conserving endangered species. In conclusion, somatic cell banking can be suggested as a tool in conservation programmes of animal genetic resources.

HER-2/neu (c-erbB-2) oncoprotein in hyperplastic endometrial polyps detected in two cats.

The presence of HER-2/neu (c-erbB-2) oncoprotein, oestrogen-alpha receptor (ER), and progesterone receptor (PR) in hyperplastic endometrial polyps (EPs) of two cats with cystic endometrial hyperplasia-pyometra (CEH-P) complex was investigated. Immunohistochemistry assay for ER, PR and c-erbB-2 oncoprotein in the glandular and stromal tissue of the EPs was performed. ER and c-erbB-2 immunoreactivity was observed in the glandular epithelium of the EPs whereas PR immunoreactivity was detected only in the stromal fibroblasts. The c-erbB-2 oncoprotein may play a role with the ER in the pathogenesis of the hyperplastic EPs, although the role of this oncoprotein in the pathogenesis of EPs has yet to be determined.

Follow-up examinations after medical treatment of pyometra in cats with the progesterone-antagonist aglepristone.

The aim of this study was to determine the therapeutic success of the medical treatment of pyometra with the antigestagen aglepristone and to document the recurrence rate in relation to the time interval after treatment with antigestagens in cats. Ten cats, 2-13 years of age and nulliparous were used in the study. The cats were treated with aglepristone at a dose of 10mg/kg body weight subcutaneously on days 1, 2, 7 and 14 (if not cured). In addition, trimethoprim/sulphadoxine was also administered at a dose of 15mg/kg body weight subcutaneously once a day for 7 days. Nine out of the 10 cats responded well to treatment. No recurrence was observed in a follow-up period of 2 years. No side effects were observed. The data suggest that aglepristone treatment is a promising approach for the medical treatment of pyometra in cats.

Tubulopapillary mammary carcinoma in a brown bear (Ursus arctos).

A 28-yr-old, nulliparous female brown bear (Ursus arctos) at the Karacabey Ovakurusu Bear Sanctuary presented with an enlargement of the mammary gland. Three other nodules were also noted in the proximity of the mammary gland and over the vulva. Clinical, hematologic, ultrasonographic, and radiologic examinations were performed; the enlarged mammary gland was removed and the other masses were also excised. Histopathologic examination revealed tubulopapillary carcinoma of the mammary gland, and the other masses were diagnosed as epidermoid cysts. This is the first reported case of tubulopapillary mammary carcinoma accompanied by epidermoid cysts in a bear.