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Simple Summary: Hypoxia drives the dysregulation of RNA molecules in canine oral melanoma (COM), influencing tumor progression and metastasis. While hypoxia-associated miRNAs have been explored, this study focused on other non-coding RNAs (ncRNAs) using NGS and qPCR. The present study revealed significant hypoxia-regulated alterations in ncRNA expression profiles, highlighting ENSCAFT00000084705.1 as consistently downregulated in COM tissues and cell lines compared to healthy tissue. Notably, its absence in plasma and extracellular vesicles suggests limited biomarker potential. This study provides evidence of transcriptional changes in ncRNAs except for miRNAs in COM, highlighting ENSCAFT00000084705.1 as a promising candidate for future investigations into the role of the transcriptome in the hypoxia-driven progression of this aggressive cancer. Hypoxia contributes to tumor progression and metastasis, and hypoxically dysregulated RNA molecules may, thus, be implicated in poor outcomes. Canine oral melanoma (COM) has a particularly poor prognosis, and some hypoxia-mediated miRNAs are known to exist in this cancer; however, equivalent data on other hypoxically dysregulated non-coding RNAs (ncRNAs) are lacking. Accordingly, we aimed to elucidate non-miRNA ncRNAs that may be mediated by hypoxia, targeting primary-site and metastatic COM cell lines and clinical COM tissue samples in next-generation sequencing (NGS), with subsequent qPCR validation and quantification in COM primary and metastatic cells and plasma and extracellular vesicles (EVs) for any identified ncRNA of interest. The findings suggest that a number of non-miRNA ncRNA species are hypoxically up- or downregulated in COM. We identified one ncRNA, the long ncRNA fragment ENSCAFT00000084705.1, as a molecule of interest due to its consistent downregulation in COM tissues, hypoxically and normoxically cultured primary and metastatic cell lines, when compared to the oral tissues from healthy dogs. However, this molecule was undetectable in plasma and plasma EVs, suggesting that its expression may be tumor tissue-specific, and it has little potential as a biomarker. Here, we provide evidence of hypoxic transcriptional dysregulation for ncRNAs other than miRNA in COM for the first time and suggest that ncRNA ENSCAFT00000084705.1 is a molecule of interest for future research on the role of the transcriptome in the hypoxia-mediated progression of this aggressive cancer. [ABSTRACT FROM AUTHOR]

Hepatocellular adenoma (HCA) and hepatocellular carcinoma (HCC) can be difficult to differentiate but must be diagnosed correctly as treatment and prognosis for these tumors differ markedly. Relevant diagnostic biomarkers are thus needed, and those identified in dogs may have utility in human medicine because of the similarities between human and canine HCA and HCC. A tRNA-derived fragment (tRF), tRNA-Val, is a promising potential biomarker for canine mammary gland tumors but has not previously been investigated in hepatic tumors. Accordingly, we aimed to elucidate the potential utility of tRNA-Val as a biomarker for canine HCA and HCC using clinical samples (tumor tissue and plasma extracellular vesicles [EVs]) and tumor cell lines with qRT-PCR assays. We also investigated relevant functions and signaling pathways with bioinformatic analyses (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes). tRNA-Val was markedly downregulated in HCC tumor tissue versus HCA tumor tissue and normal liver tissue, and a similar trend was shown in plasma EVs and HCC cell lines versus healthy controls. Based on areas under the receiver operating characteristic curves (AUCs), tRNA-Val significantly distinguished HCC (AUC = 1.00, p = 0.001) from healthy controls in plasma EVs and HCC from HCA (AUC = 0.950, p = 0.01). Bioinformatics analysis revealed that tRNA-Val may be primarily involved in DNA repair, mRNA processing, and splicing and may be linked to the N-glycan and ubiquitin-mediated proteasome pathways. This is the first report on the expression of tRNA-Val in canine HCC and HCA and its possible functions and signaling pathways. We suggest that tRNA-Val could be a promising novel biomarker to distinguish canine HCC from HCA. This study provides evidence for a greater understanding of the role played by tRNA-Val in the development of canine HCC. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background/Aim: Canine mammary gland tumors (MGTs), as a potential model of human breast cancer, have a well-defined histological classification system. MicroRNA (miRNA) expression is a key part of the molecular signatures of both MGTs and human breast cancer, although the signatures alone do not yet provide a sufficient basis for definitive diagnosis. In this study, we investigated the association between miRNA expression patterns and histological classification. Materials and Methods: Mammary gland tissue was collected from healthy dogs (n=7) and dog patients (n=80). Further samples (n=5) were obtained from established MGT cell lines. We targeted miRNAs differentially expressed in metastatic tumor tissue versus non-metastatic and normal tissue. A subset of samples was analyzed using small RNA next generation sequencing (NGS) with subsequent qPCR. Results: Six differentially expressed miRNAs were selected from the NGS analysis and submitted for large-scale qPCR. The large-scale qPCR analysis revealed greater alternations in miRNA expression. Large-scale analysis, based on 79 samples, revealed a hierarchical clustering based on selected miRNAs that did not strikingly match the histopathological subtype classification. Conclusion: We successfully investigated the large-scale miRNA expression pattern in canine MGT and provided the whole miRNA expression. The selected miRNA demonstrated that there is no straightforward mapping between molecular signatures and histological classification of canine MGTs at the miRNA level. [ABSTRACT FROM AUTHOR]

Among small non‐coding RNAs (sncRNAs/sRNAs), the functional regulation of microRNAs (miRNAs) has been studied in canine oral melanoma (COM). However, the expression level of other sncRNAs, like small nucleolar RNAs (snoRNAs), small nuclear RNAs (snRNAs), transfer RNA‐derived fragments (tRFs) and PIWI‐interacting RNAs (piRNAs), in COM is unknown. The aim of this study was to investigate sncRNAs other than miRNAs in COM from our small RNA sequencing project (PRJNA516252). We found that several snRNAs and piRNAs were upregulated, whereas tRFs and snoRNAs were downregulated in COM. Upregulation of U1 snRNA and piR‐972, and downregulation of tRNA‐ser (1) and snoRA24 was confirmed in dog melanoma tissue and cell lines by quantitative reverse transcription PCR. Consistently, the expression of tRNA‐ser (1) and snoRA24 in plasma of COM cases was also decreased. Finally, we found a similar expression trend of U1 and snoRA24 in the human cutaneous melanoma cell line, MEWO, compared with human epidermal melanocyte cells (HEMa‐Lp). In our study, snRNA, snoRNA, tRFs and piRNA were dysregulated during melanoma progression. Moreover, the melanoma‐associated expression of U1 and snoRA24 was similar in human and dog melanoma. [ABSTRACT FROM AUTHOR]

Mastitis is a common inflammatory infectious disease in dairy cows. To understand the microRNA (miRNA) expression profile changes during bovine mastitis, we undertook a genome‐wide miRNA study of normal milk and milk that tested positive on the California mastitis test for bovine mastitis (CMT+). Twenty‐five miRNAs were differentially expressed (23 miRNAs upregulated and two downregulated) during bovine mastitis relative to their expression in normal milk. Upregulated mature miR‐1246 probably derived from a U2 small nuclear RNA rather than an miR‐1246 precursor. The significantly upregulated miRNA precursors and RNU2 were significantly enriched on bovine chromosome 19, which is homologous to human chromosome 17. A gene ontology analysis of the putative mRNA targets of the significantly upregulated miRNAs showed that these miRNAs were involved in binding target mRNA transcripts and regulating target gene expression, and a Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the upregulated miRNAs were predominantly related to cancer and immune system pathways. Three novel miRNAs were associated with bovine mastitis and were relatively highly expressed in milk. We confirmed that one of the novel mastitis‐related miRNAs was significantly upregulated using a digital PCR system. The differentially expressed miRNAs were involved in human cancers, infections, and immune‐related diseases. The genome‐wide analysis of miRNA profiles in this study provides insight into bovine mastitis and inflammatory diseases. Databases: The miRNAseq generated for this study can be found in the Sequence Read Archive (SRA) under BioProject Number PRJNA421075 and SRA Study Number SRP126134 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA421075). [ABSTRACT FROM AUTHOR]

The present study was undertaken to mitigate Salmonella from betel leaf in Mymensingh. A total of 35 betel leaf samples were collected from 2 baroujes and 5 local markets in Mymensingh. The samples were sub-divided into two groups: (i) phosphate buffer solution (PBS) washed, and (ii) grinded sample. There was control and treated (with 1.5% vinegar, sorbitol, and sodium benzoate) sub-groups in both groups. Mitigation of Salmonella was determined by comparing Total Viable Count (TVC) and Total Salmonella Count (TSAC) of control with treated groups. No bacterial growth was observed in the betel leaf samples collected directly from barouj level. At market level, when grinded, there was no growth of bacteria in Plate Count Agar (PCA) and Salmonella- Shigella (SS) or Xylose Lysine De-oxychocolate (XLD) in both treated and untreated groups. But when the PBS washed samples were used, the TVC (mean log CFU±SD/mL) of betel leaf ranged from 5.16±0.82 to 5.96±1.11, whereas the TSAC value ranged from 4.87±0.58 to 5.56±1.00 for untreated group. In vinegar, there was no growth, but when treated with sorbitol, the TVC (mean log CFU±SD/mL) value reduced to 5.00±0.54 to 5.66±1.09, and TSAC (mean log CFU±SD/mL) value reduced to 4.28±0.71 to 4.78±0.64. When treated with sodium benzoate, the TVC (mean log CFU±SD/mL) value reduced to 5.06±0.53 to 5.75±1.02, and TSAC (mean log CFU±SD/mL) value reduced to 4.34±0.79 to 4.92±0.64. Data of this study indicates that all the three chemicals were effective in terms of reducing bacterial load but vinegar (1.5%) was found to be the most effective against Salmonella as well as some other bacteria when treated for 10 min. [ABSTRACT FROM AUTHOR]

Background: Menopause is a well-known risk factor for decreasing cognitive function in women. Postmenopausal women are increasing in number but relevant studies are very scarce. This study compared the cognitive function between urban and rural postmenopausal women and assessed the influence of socio-demographic factors on cognitive function., Objectives: The aim of the study was to assess the association between the cognitive function of urban and rural postmenopausal women., Methods: This comparative cross-sectional study was conducted among 87 urban and 87 rural postmenopausal women who were selected by purposive sampling method from the Nakhalpara and Dhamrai area of Dhaka district during the period from January to December 2020. Data were collected with a semi-structured questionnaire based on the Bengali version of the Mini-Mental State Examination (MMSE) scale through face-to-face interviews and record reviewing with a checklist. Statistical analyses of the results were obtained using Microsoft Excel (Microsoft Corporation, Redmond, WA) and SPSS version 24 (IBM Corp., Armonk, NY)., Results: The mean age of postmenopausal women was 58.09 ± 8.163 years in urban areas and 60.00 ± 7.562 years in rural areas. The majority (31, 35.6%) of urban women were primary school pass whereas 58 (66.7%) rural women were illiterate. The mean family income of the women was 43022.99 ± 10992.57 Bangladeshi taka (BDT) in the urban group and 14022.99 ± 5023.14 BDT in the rural group. The study revealed that 31 (35.6%) women in the urban group and 53 (60.9%) women in the rural group had abnormal cognitive function., Conclusion: The percentage of abnormal cognitive function was higher in rural postmenopausal women. Cognitive function has an association with monthly family income, housing condition, family type, age at the time of marriage, lifestyle, and co-morbidities. Policymakers can take the findings as a guide to formulate policies and programs for the improvement of cognitive function of postmenopausal women., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Husna et al.)