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Abstract Background Dihydroflavonol 4-reductase (DFR) is the key enzyme committed to anthocyanin and proanthocyanidin biosynthesis in the flavonoid biosynthetic pathway. DFR proteins can catalyse mainly the three substrates (dihydrokaempferol, dihydroquercetin, and dihydromyricetin), and show different substrate preferences. Although relationships between the substrate preference and amino acids in the region responsible for substrate specificity have been investigated in several plant species, the molecular basis of the substrate preference of DFR is not yet fully understood. Results By using degenerate primers in a PCR, we isolated two cDNA clones that encoded DFR in buckwheat (Fagopyrum esculentum). Based on sequence similarity, one cDNA clone (FeDFR1a) was identical to the FeDFR in DNA databases (DDBJ/Gen Bank/EMBL). The other cDNA clone, FeDFR2, had a similar sequence to FeDFR1a, but a different exon-intron structure. Linkage analysis in an F2 segregating population showed that the two loci were linked. Unlike common DFR proteins in other plant species, FeDFR2 contained a valine instead of the typical asparagine at the third position and an extra glycine between sites 6 and 7 in the region that determines substrate specificity, and showed less activity against dihydrokaempferol than did FeDFR1a with an asparagine at the third position. Our 3D model suggested that the third residue and its neighbouring residues contribute to substrate specificity. FeDFR1a was expressed in all organs that we investigated, whereas FeDFR2 was preferentially expressed in roots and seeds. Conclusions We isolated two buckwheat cDNA clones of DFR genes. FeDFR2 has unique structural and functional features that differ from those of previously reported DFRs in other plants. The 3D model suggested that not only the amino acid at the third position but also its neighbouring residues that are involved in the formation of the substrate-binding pocket play important roles in determining substrate preferences. The unique characteristics of FeDFR2 would provide a useful tool for future studies on the substrate specificity and organ-specific expression of DFRs.

Abstract Background Cervical cancer is the most common cancer in women living in Vientiane, Lao People’s Democratic Republic (PDR). This study examines cervical cytology using a liquid-based cytology (LBC) method and reports the presence of high-risk (HR) human papillomavirus (HPV). Methods We collected cervical samples from 1475 asymptomatic and healthy volunteers from six hospitals in Lao PDR. A total of 1422 volunteers (mean age 39.1 ± 6.4 years, range 30-54 years) were included in the final analysis. We performed HPV typing using the polymerase chain reaction technique to detect HR-HPV samples with abnormal cytology. Results The overall rates of abnormal cytology and HR-HPV–positive in the samples were 9.3% (132/1422) and 47.7% (63/132), respectively. The samples with abnormal cytology included 13 high-grade squamous intraepithelial lesions and one squamous cell carcinoma case. The results showed that the most common type of HPV was HPV16 (20.5%) followed by HPV58 (9.1%). Conclusions Healthy women in Vientiane, the capital of Lao PDR, have high rates of abnormal cervical cytology and are likely to be HR-HPV-positive. A system for detection and prevention of cervical cancer in these women should be developed in the near future.

Abstract Genomic variation includes single-nucleotide variants, small insertions or deletions (indels), and copy number variants (CNVs). CNVs affect gene expression by altering the genome structure and transposable elements within a region. CNVs are greater than 1 kb in size; hence, CNVs can produce more variation than can individual single-nucleotide variations that are detected by next-generation sequencing. Multiple system atrophy (MSA) is an α-synucleinopathy adult-onset disorder. Pathologically, it is characterized by insoluble aggregation of filamentous α-synuclein in brain oligodendrocytes. Generally, MSA is sporadic, although there are rare cases of familial MSA. In addition, the frequencies of the clinical phenotypes differ considerably among countries. Reports indicate that genetic factors play roles in the mechanisms involved in the pathology and onset of MSA. To evaluate the genetic background of this disorder, we attempted to determine whether there are differences in CNVs between patients with MSA and normal control subjects. We found that the number of CNVs on chromosomes 5, 22, and 4 was increased in MSA; 3 CNVs in non-coding regions were considered risk factors for MSA. Our results show that CNVs in non-coding regions influence the expression of genes through transcription-related mechanisms and potentially increase subsequent structural alterations of chromosomes. Therefore, these CNVs likely play roles in the molecular mechanisms underlying MSA.

Abstract Deformation of skin and muscle is essential for bringing an animated character to life. This deformation is difficult to animate in a realistic fashion using traditional techniques because of the subtlety of the skin deformations that must move appropriately for the character design. In this paper, we present an algorithm that generates natural, dynamic, and detailed skin deformation (movement and jiggle) from joint angle data sequences. The algorithm has two steps: identification of parameters for a quasi-static muscle deformation model, and simulation of skin deformation. In the identification step, we identify the model parameters using a musculoskeletal model and a short sequence of skin deformation data captured via a dense marker set. The simulation step first uses the quasi-static muscle deformation model to obtain the quasi-static muscle shape at each frame of the given motion sequence (slow jump). Dynamic skin deformation is then computed by simulating the passive muscle and soft tissue dynamics modeled as a mass–spring–damper system. Having obtained the model parameters, we can simulate dynamic skin deformations for subjects with similar body types from new motion data. We demonstrate our method by creating skin deformations for muscle co-contraction and external impacts from four different behaviors captured as skeletal motion capture data. Experimental results show that the simulated skin deformations are quantitatively and qualitatively similar to measured actual skin deformations.

Buckwheat is recognized as an important traditional crop and supports local economies in several regions around the world. Buckwheat is used, for example, as a cereal grain, noodle and bread. In addition, buckwheat is also used as a sprout or a young seedling. For these foods, sprouting is an important characteristic that affects food quality. For foods made from buckwheat flour, pre-harvest sprouting may decrease yield, which also leads to the deterioration of noodle quality. Breeding buckwheat that is resistant to pre-harvest sprouting is therefore required. Germination and subsequent growth are also important characteristics of the quality of sprouts. Although buckwheat sprouts are the focus because they contain many functional compounds, such as rutin, several problems have been noted, such as thin hypocotyls and husks remaining on sprouts. To date, several new varieties have been developed to resolve these quality issues. In this review, we summarize and introduce research on the breeding of buckwheat related to quality, sprouting and subsequent sprout growth.

Buckwheat (Fagopyrum esculentum) is recognized as an important traditional crop in some regions, and its taste is an important characteristic. Of the three cultivated buckwheat species, Tartary buckwheat (Fagopyrum tataricum) and perennial buckwheat (Fagopyrum cymosum) have strong bitterness in their seeds, which has prevented the wider use of the seeds of these varieties. In Tartary buckwheat, some studies have focused on the cause of strong bitterness generation. Tartary buckwheat seeds contain large amounts of the functional compounds rutin and rutinosidase, and rutin hydrolysis by rutinosidase has been found to be the trigger of rutin hydrolysis. Therefore, a variety with only a trace of rutinosidase and with reduced bitterness is required. The rutinosidase in Tartary buckwheat seeds consists of two major isozymes with very similar enzymatic characteristics, which can hydrolyze flour rutin within several minutes after the addition of water. Recently, the trace-rutinosidase variety Manten-Kirari in Tartary buckwheat was developed. The trace-rutinosidase characteristics were dominated by a single recessive gene. In ‘Manten-Kirari’ dough and foods, such as breads, confectionaries, and noodles, the rutin residual ratio was higher and bitterness was reduced compared to that of the normal-rutinosidase variety. In this review, we summarize the detailed research on the breeding of buckwheat related to reducing bitterness and rutin hydrolysis.