Development of PCR-based markers for identification of wheat HMW glutenin Glu-1Bx and Glu-1By alleles.

BACKGROUND: In common wheat (Triticum aestivum L.), allelic variations in the high-molecular-weight glutenin subunits Glu-B1 locus have important effects on grain end-use quality. The Glu-B1 locus consists of two tightly linked genes encoding x- and y-type subunits that exhibit highly variable frequencies. However, studies on the discriminating markers of the alleles that have been reported are limited. Here, we developed 11 agarose gel-based PCR markers for detecting Glu-1Bx and Glu-1By alleles. RESULTS: By integrating the newly developed markers with previously published PCR markers, nine Glu-1Bx locus alleles (Glu-1Bx6, Glu-1Bx7, Glu-1Bx7*, Glu-1Bx7 OE, Glu-1Bx13, Glu-1Bx14 (-) , Glu-1Bx14 (+)/Bx20, and Glu-1Bx17) and seven Glu-1By locus alleles (Glu-1By8, Glu-1By8*, Glu-1By9, Glu-1By15/By20, Glu-1By16, and Glu-1By18) were distinguished in 25 wheat cultivars. Glu-1Bx6, Glu-1Bx13, Glu-1Bx14 (+)/Bx20, Glu-1By16, and Glu-1By18 were distinguished using the newly developed PCR markers. Additionally, the Glu-1Bx13 and Glu-1Bx14 (+)/Bx20 were distinguished by insertions and deletions in their promoter regions. The Glu-1Bx6, Glu-1Bx7, Glu-1By9, Glu-1Bx14 (-), and Glu-1By15/By20 alleles were distinguished by using insertions and deletions in the gene-coding region. Glu-1By13, Glu-1By16, and Glu-1By18 were dominantly identified in the gene-coding region. We also developed a marker to distinguish between the two Glu-1Bx14 alleles. However, the Glu-1Bx14 (+) + Glu-1By15 and Glu-1Bx20 + Glu-1By20 allele combinations could not be distinguished using PCR markers. The high-molecular-weight glutenin subunits of wheat varieties were analyzed by ultra-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the findings were compared with the results of PCR analysis. CONCLUSIONS: Seven Glu-1Bx and four Glu-1By allele detection markers were developed to detect nine Glu-1Bx and seven Glu-1By locus alleles, respectively. Integrating previously reported markers and 11 newly developed PCR markers improves allelic identification of the Glu-B1 locus and facilitates more effective analysis of Glu-B1 alleles molecular variations, which may improve the end-use quality of wheat.

Exposure to Hazardous Substances and Their Health Effects Among Residents Living Near Three Industrial Waste Incinerators in Korea.

BACKGROUND: This study aimed to evaluate exposure to various hazardous substances emitted by incineration facilities and their likely effect on the health for residents of Bugi-myeon, Cheongju, Korea, which has three incineration facilities. METHODS: Heavy metals, polycyclic aromatic hydrocarbons (PAHs), and dioxin concentrations in the air and soil of exposed and control areas were measured. Moreover, the exposure levels to harmful substances and its effects on health were investigated in 1,124 exposed and 232 control adults. RESULTS: PAHs and dioxin concentrations in the air in the exposed area were significantly higher than in the control area. Urinary cadmium and PAHs metabolite concentrations were significantly higher in the exposed group than in the control group. The exposure group also had a higher prevalence of depression and self-reported allergic symptoms than the control group. CONCLUSION: The possibility of residents in Bugi-myeon being exposed to hazardous substances at incineration facilities cannot be ruled out. To prevent them from further exposure to hazardous substances, it is necessary to prohibit the expansion of additional incineration facilities in this area and to implement continuous monitoring projects for residents.

Cancer risk in the residents of a town near three industrial waste incinerators in Korea: a retrospective cohort study.

BACKGROUND: Three industrial waste incinerators (IWIs) were built in 1999, 2001, and 2010, within a 3 km radius of a town with a population of around 5000 in Korea. This study evaluated whether residents near these three IWIs had increased cancer incidence than those from other areas in Korea using regional health data. METHODS: Standardized incidence ratios (SIR) were calculated using the frequency of cancer cases in the National Cancer Registry of the exposed area (Buki-myeon), Chungcheongbuk-do (Chungbuk, state including Buki-myeon), and whole Korea from 1999 to 2017. A retrospective cohort was created using National Health Insurance System data from 2002 to 2018. The exposed group was defined as those having a residential history in the exposed area. The control group was defined as those having a residential history in nearby towns or counties in Chungbuk, excluding counties having living and cultural areas in other provinces and cities. Hazard ratio (HR) and 95% confidence intervals (CI) were estimated using the Cox proportional hazard model adjusted for age, level of health insurance fee, and smoking history. RESULTS: In the ecological study using National Cancer Registry data, the risk of all cancers, all cancers excluding thyroid, esophageal, stomach, and lung cancers in the exposed area were 1.13 (95% CI 1.03-1.24), 1.15 (95% CI 1.04-1.26), 1.91 (95% CI 1.13-2.89), 1.39 (95% CI 1.14-1.66), and 1.29 (95% CI 1.03-1.57) times higher than in whole Korea among exposed males, respectively. In the retrospective cohort, 4300 males (26,821 person-years) and 3796 females (24,746 person-years) in exposed group, 150,964 males (1,212,010 person-years) and 134,535 females (1,104,025 person-years) in control group were analyzed. After adjusting for several confounding factors, the risks for gallbladder cancer among males and kidney cancer among females were 2.65 (95% CI 1.38-5.06) and 2.82 (95% CI 1.13-7.03) times higher in the exposed group versus the control group, respectively. CONCLUSIONS: Cancer risk was higher in Koreans having residential history living near IWIs compared to the other areas. Further study warrants nationwide effects and longer follow-up of WIs for cancers in Korea.

Cancer cluster among small village residents near the fertilizer plant in Korea.

OBJECTIVES: In Jang-jeom, a small village in Hamra-myeon, Iksan-si, Jeollabuk-do, South Korea, residents raised concerns about a suspected cancer cluster that they attributed to a fertilizer plant near the village. We aimed to investigate whether the cancer incidence in the village was higher than that in the general Korean population when the factory was in operation (2001-2017) and whether living in the village was associated with a higher risk of cancer. METHODS: Using national population data and cancer registration data of South Korea, we estimated the standardized incidence ratios (SIRs) in the village to investigate whether more cancer cases occurred in the village compared to other regions. The SIRs were standardized by age groups of 5 years and sex. In order to investigate whether residence in the village increased the risk of cancer, a retrospective cohort was constructed using National Health Insurance Service (NHIS) databases. We estimated the cancer hazard ratios (HRs) using the Cox proportional hazard model, and defined the exposed area as the village of Jang-jeom, and the unexposed or control area as the village neighborhood in Hamra-myeon. We considered potential confounding variables such as age, sex, and income index in the models. Additionally, we measured polycyclic aromatic hydrocarbons (PAHs) and tobacco-specific nitrosamines (TSNAs), suspected carcinogens that may have caused the cancer cluster, in samples collected from the plant and the village. RESULTS: Twenty-three cancer cases occurred in Jang-jeom from 2001 to 2017. Between 2010 and 2016, the incidence rates of all cancers (SIR: 2.05, except thyroid cancer: 2.22), non-melanoma skin cancer (SIR: 21.14, female: 25.41), and gallbladder (GB) and biliary tract cancer in men (SIR: 16.01) in the village were higher than those in the national population in a way that was statistically significant. In our cohort analysis that included only Hamra-myeon residents who have lived there for more than 7 years, we found a statistically significant increase in the risk of all cancers (HR: 1.99, except thyroid cancer: 2.20), non-melanoma skin cancer (HR: 11.60), GB and biliary tract cancer (HR: 15.24), liver cancer (HR: 6.63), and gastric cancer (HR: 3.29) for Jang-jeom residents compared to other Hamra area residents. We identified PAHs and TSNAs in samples of deposited dust and residual fertilizer from the plant and TSNAs in dust samples from village houses. CONCLUSIONS: The results of the SIR calculation and cancer risk analyses of Jang-jeom village residents from the retrospective cohort design showed consistency in the effect size and direction, suggesting that there was a cancer cluster in Jang-jeom. This study would be a good precedent for cancer cluster investigation.

Loss-of-Function Alleles of Heading date 1 (Hd1) Are Associated With Adaptation of Temperate Japonica Rice Plants to the Tropical Region.

Adaptation of temperate japonica rice varieties to tropical regions is impeded by extremely early flowering probably due to photoperiod change from long to short. However, constant breeding efforts led to development of temperate japonica varieties adapted to tropical/subtropical regions, but the genetic factor underlying this is still elusive. We analyzed the 45 diverse rice accessions and 12 tropical-adapted temperate japonica lines for the allele types of seven major flowering genes Hd1, OsPPR37, DTH8, Ghd7, Ehd1, RFT1, and Hd3a and flowering time under three different field conditions in temperate and tropical locations. The accessions originated from the tropical/subtropical regions preferred the non-functional alleles of Hd1 and not other flowering genes. The genetic effect analysis of each gene showed that only the functional Hd1 caused early flowering in the tropical location. All 12 temperate japonica breeding lines adapted to the tropics possessed the loss-of-function alleles of Hd1 with no change of other flowering genes compared to common Korean temperate japonica varieties. A phylogenetic analysis using 2,918 SNP data points revealed that the genome status of the 12 breeding lines were very similar to Korean temperate japonica varieties. These results indicate that the functional Hd1 alleles of temperate japonica varieties induced extremely early flowering in the tropics and the non-functional hd1 alleles brought about the adaptation of temperate japonica rice to tropical regions.

Sugary Endosperm is Modulated by Starch Branching Enzyme IIa in Rice (Oryza sativa L.).

BACKGROUND: Starch biosynthesis is one of the most important pathways that determine both grain quality and yield in rice (Oryza sativa L.). Sugary endosperm, sugary-1 (sug-1), is a mutant trait for starch biosynthesis. Rice plants carrying sug-1 produce grains that accumulate water-soluble carbohydrates instead of starch, even after maturity. Although this trait enhances the diversity of grain quality, sugary endosperm rice has hardly been commercialized due to the severely wrinkled grains and subsequent problems in milling. This study was conducted to identify the genes responsible for the sug-h phenotype through a map-based cloning technology. RESULTS: We induced a mild sugary mutant, sugary-h (sug-h) through the chemical mutagenesis on the Korean japonica cultivar Hwacheong. Grains of the sug-h mutant were translucent and amber-colored, and the endosperm appeared less wrinkled than sug-1, whereas the soluble sugar content was fairly high. These characteristics confer greater marketability to the sug-h mutant. Genetic analyses indicated that the sug-h mutant phenotype was controlled by a complementary interaction of two recessive genes, Isoamylase1 (OsISA1), which was reported previously, and Starch branching enzyme IIa (OsBEIIa), which was newly identified in this study. Complementation tests indicated that OsBEIIa regulated the properties of sugary endosperm. CONCLUSIONS: Complementary interactions between the starch biosynthesis genes OsISA1 and OsBEIIa determine the mild sugary endosperm mutant, sugary-h, in rice. Our finding may facilitate the breeding of sugaryendosperm rice for commercial benefit.

Morphological and starch characteristics of the Japonica rice mutant variety Seolgaeng for dry-milled flour.

Producing good-quality, fine rice flour is more difficult than wheat flour because the rice grain is harder. The non-glutinous Japonica-type variety Seolgaeng, derived from N-methyl-N-nitrosourea (MNU) mutagenesis, and four other varieties, representing a range of amylose contents, were evaluated in this study. Dry-milled Seolgaeng rice flour exhibited an average particle size that is <70 µm, a more uniform particle-size proportion than other varieties. Moreover, we noted significant differences in the damaged starch content in flour from Seolgaeng compared to the other varieties (p<0.05). Seolgaeng flour showed a round starch structure, which would lead to better friability, finer particle size, and less damage to the endosperm during dry milling. Indeed, among all varieties evaluated in this study, dry-milled Seolgaeng flour had the finest particle size (averaging <70 µm) and exhibited less damaged starch. With its round starch granules, Seolgaeng is a suitable candidate for drymilled rice flour.

Characteristics of illegal and legal cigarette packs sold in Guatemala.

BACKGROUND: Guatemala, as a party to the Framework Convention on Tobacco Control (FCTC), is required to regulate cigarette packaging and labeling and eliminate illicit tobacco trade. Current packaging and labeling characteristics (of legal and illegal cigarettes) and their compliance with the FCTC is unknown. METHODS: We sought to analyze package and label characteristics of illegal and legal cigarettes sold in Guatemala. We visited the 22 largest traditional markets in the country to purchase illegal cigarettes. All brands registered on tobacco industry websites were purchased as legal cigarettes. Analysis compared labeling characteristics of illegal and legal packs. FINDINGS: Most (95%) markets and street vendors sold illegal cigarettes; 104 packs were purchased (79 illegal and 25 legal). Ten percent of illegal and none of the legal packs had misleading terms. Half of the illegal packs had a warning label covering 26 to 50% of the pack surface. All legal packs had a label covering 25% of the surface. Illegal packs were more likely to have information on constituents and emissions (85% vs. 45%, p < 0.001) and were less expensive than legal ones (USD 0.70 ± 0.7 and 1.9 ± 1.8, p < 0.001). CONCLUSIONS: In Guatemala, neither illegal nor legal cigarette packs comply with FCTC labeling mandates. Urgent implementation and enforcement of the FCTC is necessary to halt the tobacco epidemic.

The Effects of Mind Subtraction Meditation on Depression, Social Anxiety, Aggression, and Salivary Cortisol Levels of Elementary School Children in South Korea.

This study analyzed the effects of a school-based mind subtraction meditation program on depression, social anxiety, aggression, and salivary cortisol levels of 42 elementary school children in South Korea. The research design was a nonequivalent group comparison with pretest and post-test. The experimental group was given 8weeks of the meditation program. The results showed social anxiety, aggression, and salivary cortisol levels were significantly lowered in the experimental group. This demonstrated that the school-based mind subtraction meditation program could be effective in improving psychosocial and behavioral aspects of mental health in elementary school children.

Physicochemical and Gelatinization Properties of Starches Separated from Various Rice Cultivars.

UNLABELLED: Morphological, viscoelastic, hydration, pasting, and thermal properties of starches separated from 10 different rice cultivars were investigated. Upon gelatinization, the G' values of the rice starch pastes ranged from 37.4 to 2057 Pa at 25 °C, and remarkably, the magnitude depended on the starch varieties. The rheological behavior during gelatinization upon heating brought out differences in onset in G' and degree of steepness. The cultivar with high amylose content (Goami) showed the lowest critical strain (γ(c)), whereas the cultivars with low amylose content (Boseokchal and Shinseonchal) possessed the highest γ(c). The amylose content in rice starches affected their pasting properties; the sample possessing the highest amylose content showed the highest final viscosity and setback value, whereas waxy starch samples displayed low final viscosity and setback value. The onset gelatinization temperatures of the starches from 10 rice cultivars ranged between 57.9 and 64.4 °C. The amylose content was fairly correlated to hydration and pasting properties of rice starches but did not correlate well with viscoelastic and thermal characteristics. The combined analysis of hydration, pasting, viscoelastic, and thermal data of the rice starches is useful in fully understanding their behavior and in addressing the processability for food applications. PRACTICAL APPLICATION: Rice flour has potential applications in various food products. The physicochemical properties of rice flour are dependent on its variety, which affects the quality of the final products. In this study, the combined analysis including hydration, pasting, viscoelastic, and thermal properties of rice flour could afford information for preparing a particular product such as bread and noodle.

Changes in lipid substances in rice during grain development.

Lipid substances, such as fatty acids, γ-oryzanol, policosanols, and tocol (tocopherol+tocotrienol), were investigated in two rice cultivars, Ilpum and Dasan, during rice development. In both cultivars, the lipid extract level decreased steadily after it reached its maximum level. Additionally, there were differences in the fatty acid composition, depending on the rice developmental stage, between the two cultivars. The γ-oryzanol level increased during rice development, and there were differences in the composition of γ-oryzanol between the two cultivars. The levels of policosanols drastically decreased during the early stage of rice development in the two cultivars. The total tocol level showed a downward trend during rice development. The predominant tocol isomer in Ilpum was α-tocopherol during rice development. In Dasan, the predominant tocol isomer was α-tocopherol at the early stage, but γ-tocotrienol at the later stage. This study provided information on the levels and composition of lipid substances, such as fatty acids, γ-oryzanol, policosanols, and tocol during rice development.

Preparation of highly purified pinolenic acid from pine nut oil using a combination of enzymatic esterification and urea complexation.

Pinolenic acid (PLA) is a polyunsaturated fatty acid of plant origin. PLA has been successfully enriched according to a two-step process involving lipase-catalysed esterification and urea complexation. For the first step, the fatty acids present in pine nut oil were selectively esterified with lauryl alcohol using Candida rugosa lipase. Under the optimum conditions of 0.1% enzyme loading, 10% additional water, and 15 °C, PLA was enriched up to 43 mol% from an initial value of 13 mol% in the pine nut oil. For the second step, the PLA-enriched fraction from the first step was subjected to a urea complexation process. In this way, PLA enrichments with purities greater than 95 mol% were obtained at urea to fatty acid ratios greater than 3:1 (wt/wt), and 100% pure PLA was produced at a urea to fatty acid ratio of 5:1 with an 8.7 mol% yield.