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Simple Summary: Several DNA tests are available for individual identification and mitochondrial diseases. Among them, the sequencing of whole mitochondrial DNA (mtDNA) is a beneficial method. Although DNA analysis requires a sample from the individual dog (e.g., blood or tissue), fecal samples are considered non-invasive and the least stressful form of sampling. In this study, we attempted to establish a method for sequencing whole mtDNA using small fecal samples (100 mg) from two domestic dogs. We also attempted to establish a method that is as accessible as possible to veterinary researchers. As a result, we were able to determine the samples' whole mtDNA sequences with high accuracy. The method developed in this study shows that mtDNA from feces can be decoded and that this form of sampling provides a suitable sample for mtDNA decoding. The presented findings will inform future developments in veterinary medicine and animal welfare. Medical care for domestic dogs is now respected worldwide as being at a similar level to that of humans. We previously established a test method to determine whole mitochondrial DNA (mtDNA) using oral mucosal DNA that may be useful for medical care and welfare. However, the sample types tested in dogs are not limited to those obtained from the oral mucosa. Therefore, in the present study, we attempted to establish a test method to determine whole mtDNA sequences using feces, which represents the least invasive specimen. Two Japanese domestic dogs were used in the present study. DNA was extracted from approximately 100 mg of fresh feces from each dog, and PCRs were performed using four primer pairs that can amplify whole mtDNA. Following PCR, amplicons were pooled to create a DNA library using an experimental robot with an original program. Data were then acquired via NGS and data analysis was performed. The results showed that the whole mtDNA sequence of the two dogs was determined with high accuracy. Our results suggest that feces can be adapted for mitochondrial disease and individual identification testing and could serve as a useful testing method for the future medical care and welfare of domestic dogs. [ABSTRACT FROM AUTHOR]

The Cornus kousa tree, which is of Asian origin, is often cultivated for ornamental purposes and used in traditional medicine. The tree produces sugar-rich fruits, which are potential habitats for natural yeasts. The identification of new yeast strains has many advantages for the industry and research. This study aimed to isolate and identify yeast species from C. kousa fruits and to understand their microbial ecology. Ripe and rotten fruits, which had fallen on the ground naturally, were collected and soaked in culture media, followed by plate spreading for colony growth. The morphological examination revealed three distinct colony types, including two from the ripe fruits and one from the rotten fruits. The analysis of the internal transcribed spacer 1 region indicated three yeast strains corresponding to the three colony types: Torulaspora delbrueckii and Pichia kluyveri from the ripe fruits and Saccharomyces cerevisiae from the rotten fruits. The metabolic characterizations demonstrated that all three yeasts efficiently consumed glucose and produced alcohol. S. cerevisiae exhibited the strongest fermentation ability and the highest growth rate. These findings showed that Cornus kousa fruit is a source of diverse yeast species, with distinct species associated with different states of fruit decomposition. [ABSTRACT FROM AUTHOR]

Background: Reduction of blood group ABO antigens on red blood cells (RBCs) is well known in patients with leukemias, and this reduction of ABO expression is strongly associated with DNA methylation of the ABO promoter. Previously, we reported a two‐nucleotide deletion in RUNX1 encoding an abnormally elongated protein lacking the trans‐activation domain in a patient with myelodysplastic syndrome (MDS) showing A‐antigen loss on RBCs. This prompted us to investigate the underlying mechanism responsible for A‐antigen reduction on RBCs in another patient with MDS. Study Design and Methods: Screening of somatic mutations was carried out using a targeted sequencing panel with genomic DNA from peripheral blood mononuclear cells from the patient and eleven MDS controls without A‐ or B‐antigen loss. DNA methylation of the ABO promoter was examined by bisulfite genomic sequencing. Transient transfection assays were performed for functional evaluation of mutations. Results: Screening of somatic mutations showed missense mutations in RUNX1 and GATA2 in the patient, while no mutation was found in exons of those genes in the controls. There was no significant difference in ABO promoter methylation between the patient and the controls. Transient transfection experiments into COS‐7 and K562 cells suggested that the amino acid substitutions encoded by those mutations reduced or lost the trans‐activation potential of the ABO expression. Conclusion: Considering the discrepancy between the variant frequencies of these mutations and the ratios of the RBCs with A‐antigens loss, the antigen reduction might be associated with these somatic mutations and hypermethylation of the ABO promoter. [ABSTRACT FROM AUTHOR]