Upregulation and functional roles of miR-450b in canine oral melanoma.

Canine oral melanoma (COM) is a common and highly aggressive disease with the potential to model human melanomas. Dysregulated microRNAs represent an interesting line of research for COM because they are implicated in tumor progression. One example is miR-450b, which has been investigated for its molecular mechanisms and biological functions in multiple human cancers, but not human or canine melanoma. Here, we aimed to investigate miR-450b as a potential diagnostic biomarker of COM and its functional roles in metastatic and non-metastatic forms of the disease. We investigated the expression of miR-450b and its target mRNA genes in clinical (tumor tissue and plasma) samples and metastatic and primary-tumor cell lines. Knockdown and overexpression experiments were performed to determine the influence of miR-450b on cell proliferation, migration, colony formation, and apoptosis. miR-450b was significantly upregulated in COM and differentiated between metastatic and non-metastatic tumors, and its potential as a biomarker of metastatic and non-metastatic COM was further confirmed in ROC analysis. miR-450b knockdown promoted cell proliferation, migration, and clonogenicity and inhibited apoptosis, whereas its overexpression yielded the reverse pattern. miR-450b directly binds 3' UTR of PAX9 mRNA and modulates its function leading to BMP4 downregulation and MMP9 upregulation at the transcript level. Furthermore, we surmised that miR-450b activates the Wnt signaling pathway based on gene ontology and enrichment analyses. We concluded that miR-450b has the potential as a diagnostic biomarker and could be a target candidate for COM treatment.

Elevated expression of miR-301a and its functional roles in canine oral melanoma.

miR-301a is one of numerous dysregulated microRNAs (miRNAs) in canine oral melanoma (COM), one of which is miR-301a (upregulated). Its biological role has been described in various human cancer types, including malignant melanoma, but not in COM. Accordingly, in this study, we investigated miR-301a expression in COM in greater detail to ascertain whether it could serve as a diagnostic biomarker, elucidate its functional roles in this cancer, and predict the possible pathways by which it exerts its effects. Relative expression of miR-301a was investigated in clinical oral tissue and plasma samples and COM cell (KMeC and LMeC) lines using qRT-PCR. Knockdown of miR-301a was also validated for KMeC and LMeC cells using qRT-PCR. We performed CCK-8 assays to assess cell proliferation, monolayer wound-healing, and transwell migration assays to assess cell migration, a colony-formation assay to assess clonogenicity, a TUNEL assay and flow cytometry to assess apoptosis-related effects, and gene enrichment analyses to predict possible related pathways. miR-301a was markedly upregulated in COM oral tissue and plasma clinically, suggesting its potential as a diagnostic biomarker for COM diagnosis. In vitro assays demonstrated that miR-301 significantly inhibited apoptosis in COM cells while promoting cell migration, proliferation, and clonogenicity. We also predicted that miR-301 exerts cancer-promoting effects through the Wnt signalling pathway for COM. Our findings suggest that miR-301a is a COM oncomiR that regulates several oncogenic phenotypes with the potential to be a diagnostic biomarker.

Prevalence and Characterization of Dental and Skull-Bone Pathologies of the Raccoon Dog (Nyctereutes procyonoides) in Lithuania.

The present investigation endeavours to discern dental and non-dental pathologies affecting cranial structures of raccoon dogs, while focusing on cases of periodontitis. Furthermore, the study aims to conduct a comparative analysis based on sex and the nature of the pathologies encountered. The number of investigated skulls amounted to 126, including 76 males and 50 females. The predominant pathology identified was hypodontia, which accounted for 26.7% of males and 20% of females. Notably, the majority of hypodontia cases involved the absence of the mandibular third molar. Another noteworthy pathology was various stages of periodontitis, with rates ranging from 21.3% in males to 8% in females. Other pathologies, like tooth fractures and abrasion, were significantly less encountered. Excessive bone formation was relatively abundant and localized in specific areas-the parietal bone and the occipital regions. This tendency was observed in 8% of male cases and 6% of females. We found that the total number of dental and skull-bone pathologies is significantly more common in males than in females (p = 0.003). Additionally, the total number of various cases of periodontitis is more common in males too (p = 0.04).

microRNAs Are Abundant and Stable in Platelet-Rich Fibrin and Other Autologous Blood Products of Canines.

Various microRNAs (miRNAs) present in autologous blood products of canines have not been studied recently. We aimed to elucidate the existence of miRNAs in platelet-rich fibrin (PRF) and the stability of canine autologous blood products under various storage conditions. Total RNAs were isolated from PRF and other autologous blood products following newly adapted protocols used in commercial kits for plasma and tissue samples. Quantitative real-time polymerase chain reaction analysis (qPCR) was used to detect miRNAs in autologous blood products. The miR-16, miR-21, miR-155, and miR-146a were abundant in PRF and other autologous blood products of canines. Furthermore, we found they could maintain stability under protracted freezing temperatures of -30 °C for at least one month. Our findings revealed that PRF might be a stable resource for various canine miRNAs.

Peripheral mononuclear cells composition in platelet-rich fibrin in canines with chronic conditions.

Platelet-rich fibrin (PRF) is a hot research topic because of its regenerative effect in humans. However, data reporting about its application in companion animals is lacking. The study aimed to supplement currently available data on PRF cell composition in canine patients by isolating peripheral blood mononuclear cells (PBMC), namely T cells, matured B cells, monocytes and macrophages, and adapting current protocols of cell flow cytometry for PRF analysis. The canine patient population was divided into three subgroups: animals with periodontitis only, animals with neoplasia and periodontitis, and healthy controls. Individual clinical parameters of the patients and evaluation of the wound healing quality were included in the research. In the present study, canine PRF cell composition was analyzed for the first-time using cell flow cytometry protocol. A higher proportion of PBMC cells related to wound healing (CD3+, CD3+ CD4+ CD8-, CD14+) were found in the PRF of control, periodontitis and neoplasia groups compared to the respective blood samples, which implies a positive outcome associated with clinical PRF usage in canine patients. Proportions of monocytes and macrophages were higher in PRF samples compared to the blood of healthy patients and periodontitis-affected patients. However, inflammatory and neoplastic processes do not affect the distribution of PBMC.

Morphometric analysis of the red fox molar teeth in Lithuania.

The objective of the study was to investigate the lengths and widths of maxillary (P4 , M1 and M2 ) and mandibular (M1 , M2 and M3 ) molar teeth, lengths of dental rows and dental features of sexual dimorphism in the red fox. The correlation analysis between teeth measurements and dental row length was included too. The osteometric analysis was performed by the A. von den Driesch (1976) method. A total of 230 skulls and mandibles were investigated (n = 230): 113 males (n = 113) and 117 females (n = 117). The findings of the research revealed that most dimensions are higher in males compared with females (p < 0.001). Contrarily, Wilks' Lambda analysis showed results in the interval from 0.821 to 1, which leads to the conclusion that a teeth measurement study is unreliable to determine the sex of the red fox. The trustworthy measurements were the length of the whole dental arcade in the maxilla and mandible (UDR and LDR); Wilks' Lambda UDR = 0.860; LDR = 0.821; p < 0.001). Evaluation of the correlation between measurements revealed that results vary from very weak (0.047) to moderate strength (0.543). There were no strongly correlating measurements found. The correlation between the upper and lower molars with the length of the teeth arcade is quite weak, and only solitary measurements (M2 , L and B) had a weak correlation.