Cross species/genera transferability of simple sequence repeat markers, genetic diversity and population structure analysis in gladiolus (Gladiolus × grandiflorus L.) genotypes.

Background: Genetic analysis of gladiolus germplasm using simple sequence repeat (SSR) markers is largely missing due to scarce genomic information. Hence, microsatellites identified for related genera or species may be utilized to understand the genetic diversity and assess genetic relationships among cultivated gladiolus varieties. Methods: In the present investigation, we screened 26 genomic SSRs (Gladiolus palustris, Crocus sativus, Herbertia zebrina, Sysirinchium micranthum), 14 chloroplast SSRs (Gladiolus spp., chloroplast DNA regions) and 25 Iris Expressed Sequence Tags (ESTs) derived SSRs across the 84 gladiolus (Gladiolus × grandiflorus L.) genotypes. Polymorphic markers detected from amplified SSRs were used to calculate genetic diversity estimates, analyze population structure, cluster analysis and principal coordinate analysis (PCoA). Results: A total of 41 SSRs showed reproducible amplification pattern among the selected gladiolus cultivars. Among these, 17 highly polymorphic SSRs revealed a total of 58 polymorphic alleles ranging from two to six with an average of 3.41 alleles per marker. Polymorphic information content (PIC) values ranged from 0.11 to 0.71 with an average value of 0.48. A total of 4 SSRs were selectively neutral based on the Ewens-Watterson test. Hence, 66.66% of Gladiolus palustris, 48% of Iris spp. EST, 71.42% of Crocus sativus SSRs showed cross-transferability among the gladiolus genotypes. Analysis of genetic structure of 84 gladiolus genotypes revealed two subpopulations; 35 genotypes were assigned to subpopulation 1, 37 to subpopulation 2 and the remaining 12 genotypes could not be attributed to either subpopulation. Analysis of molecular variance indicated maximum variance (53.59%) among individuals within subpopulations, whereas 36.55% of variation among individuals within the total population. The least variation (9.86%) was noticed between two subpopulations. Moderate (FST = 0.10) genetic differentiation between two subpopulations was observed. The grouping pattern of population structure was consistent with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on simple matching dissimilarity coefficient and PCoA. Conclusion: SSR markers from the present study can be utilized for cultivar identification, conservation and sustainable utilization of gladiolus genotypes for crop improvement. Genetic relationships assessed among the genotypes of respective clusters may assist the breeders in selecting desirable parents for crossing.

Characterization of Anthocyanins and Their Antioxidant Activities in Indian Rose Varieties (Rosa × hybrida) Using HPLC.

The present study was designed to explore the anthocyanin profile and antioxidant activities in Indian rose varieties (Rosa × hybrida). Among fifty varieties, Ashwini recorded the highest total phenolic content (427.59 ± 3.47 mg GAE/100 g) along with the highest FRAP (397.15 ± 0.82 µmol trolox/g) and DPPH free radical scavenging activity (93.47 ± 0.19%) on a fresh weight basis. A significant positive correlation was observed between total anthocyanin content, total phenolic content, and antioxidant activities. Four distinct clusters were formed according to total anthocyanins, total phenols, and antioxidant activities; white- and yellow-colored varieties were most distant from red ones. Principal component analysis revealed that variable total anthocyanin content contributed to the maximum variation among the fifty rose varieties studied. Highly anthocyanin-rich rose varieties were characterized by high-performance liquid chromatography coupled with a photodiode array detector (HPLC-PAD), which identified two major components of anthocyanins, i.e., cyanidin 3,5-di-O-glucoside and pelargonidin 3,5-di-O-glucoside. Cyanidin 3,5-di-O-glucoside was the predominant anthocyanin in red- and pink-colored varieties, whereas pelargonidin 3,5-di-O-glucoside was the major one in the orange variety. The maximum cyanidin 3,5-di-O-glucoside content was recorded in variety Ashwini (497.79 mg/100 g), whereas the maximum pelargonidin 3,5-di-O-glucoside content was recorded in Suryakiran (185.43 mg/100 g). It is suggested that the rose varieties with high anthocyanin content and antioxidant activity can be exploited as a potential source of nutraceuticals in the food industry.

Production of Haploids and Doubled Haploids in Marigold (Tagetes spp.) Using Anther Culture.

Homozygous parental lines are indispensable for commercial hybrid seed production in many ornamental and vegetable crops. The in vitro induction of haploids and doubled haploids (DHs) through gametic embryogenesis is an effective approach for single-step development of complete homozygous lines from heterozygous donor plants. Anther culture is one of the most popular and widely employed techniques for development of haploids. Here we describe the detailed protocol for rapid and successful induction of haploids in Tagetes spp. using in vitro androgenesis approach. In this protocol, we have provided the comprehensive details of various steps of anther culture in marigold right from the growing of donor plants, selection of buds, pretreatment, embryogenesis and regeneration to ploidy analysis, and chromosome doubling for development of DHs.

Are Dental Patients Concerned About Safety? An Exploratory Study.

OBJECTIVES: This study addresses a gap in the literature regarding dental patients' perceptions about safety at the dental office and their attitudes toward reporting safety concerns and experiences. METHODS: We conducted a cross-sectional study with adult dental patients at an academic dental institution over a 6-week study period. A 16-item questionnaire was distributed to the patients to assess (1) past safety concerns and experiences during dental visits; (2) factors affecting the future reporting of safety concerns and experiences; (3) overall concern about safety at the dental office; (4) overall perceptions that patients should report of safety concerns or experiences to dental providers and staff. RESULTS: A majority (63.5%) of dental patients were concerned about safety at the dental office, although only one-third of them shared their past safety concerns or experiences with their dental providers or clinic staff. Irrespective of their past experiences, most patients (96.9%) believed that patients should report any safety concerns or experiences to the clinic. Being female, highly educated, and having poor oral health were associated with a decreased overall perception that patients should report safety concerns and experiences to dental care providers and staff. CONCLUSIONS: Our findings suggest that dental patients are concerned about safety and can be valuable sources of data, when adequately engaged. The current level of patient reporting of safety concerns and/or experiences to clinic staff or care providers is not optimal for learning and improvement. PRACTICAL IMPLICATIONS: Better patient engagement in safety activities will potentially increase our collective understanding of threats to safety. Therefore, dental clinics need to encourage patients to speak up about their safety concerns or experiences.

Characterization of Indian bred rose cultivars using morphological and molecular markers for conservation and sustainable management.

Rose (Rosa × hybrid L.) is one of the most important commercial ornamental crops cultivated worldwide for its beauty, fragrance and nutraceutical values. Characterization of rose germplasm provides precise information about the extent of diversity present among the cultivars. It also helps in cultivar identification, intellectual property right protection, variety improvement and genetic diversity conservation. In the present study, 109 Indian bred rose cultivars were characterized using 59 morphological and 48 SSR markers. Out of 48 SSRs used, 31 markers exhibited polymorphism and 96 alleles were identified with an average of 3.9 alleles per locus. Nei's expected heterozygosity value of each locus ranged from 0.08 (with SSR ABRII/RPU32) to 0.78 (SSR Rh58). The similarity coefficient values ranged from 0.42 to 0.90 which indicated presence of moderated diversity among Indian cultivars. The neighbor-joining tree based on morphological data grouped the cultivars into two major clusters and several minor clusters based on their morphological resemblance. However, UPGMA dendrogram constructed using matching coefficient values grouped the cultivars into eight different clusters. Interpopulation analysis revealed higher genetic similarities between Hybrid Tea and Floribunda cultivars. An analysis for presence of population sub-structure grouped the Indian cultivars into eight different genetic groups. Analysis of molecular variance revealed apportioning of 97.59% of the variation to within subgroup diversity and 3.07% to between the cultivar groups. We have demonstrated here successful utilization of robust SSR to distinguish cultivars and assess genetic diversity among Indian bred rose cultivars. The information provided here is useful for cultivar identification and protection, cultivar improvement and genetic diversity conservation.

Number of Pregnant Women at Four Dental Clinics and the Care They Received: A Dental Quality eMeasure Evaluation.

Process-of-care quality measure research can be used to identify gaps in the delivery of dental services to pregnant patients. The aim of this study was to evaluate the types of dental services that pregnant patients received in four dental clinics over five years as documented in the electronic health record (EHR). To accomplish this objective, the authors modified and validated a previously published claims-based dental quality measure for EHR use. After the electronic dental quality measure specifications were defined, the number of pregnant patients was calculated at three academic dental institutions and one large accountable care organization, and the types of dental care services they received over a five-year period (2013-17) were determined. Calibrated reviewers at each institution independently reviewed a sub-sample of patient charts to validate the information obtained from EHR queries, and the concordance between manual chart reviews and EHR query reports was analyzed. Of the 335,078 women aged 15-44 years who received care at the four clinics for the five reporting years, 3.9% (n=13,026) were pregnant. Among these pregnant patients, 48.9% (n=6,366) received a periodic dental examination; 30.0% (n=3,909) received a comprehensive dental exam; and 21.5% (n=2,799) received additional dental services, irrespective of comprehensive or periodic oral evaluations. Overall, the mean proportion of pregnant patients seeking care in these academic dental and group practice clinics was low, but 78.9% of them received either a periodic or comprehensive oral evaluation. Given the importance of oral health care during pregnancy, these findings suggest a need for curriculum development to incorporate prenatal oral health education in the training of dental students.

Unintended consequences and challenges of quality measurements in dentistry.

BACKGROUND: In recent years, several state dental programs, researchers and the Dental Quality Alliance (DQA) have sought to develop baseline quality measures for dentistry as a way to improve health outcomes, reduce costs and enhance patient experiences. Some of these measures have been tested and validated for various population groups. However, there are some unintended consequences and challenges with quality measurement in dentistry as observed from our previous work on refining and transforming dental quality measures into e-measures. MAIN BODY: Some examples of the unintended consequences and challenges associated with implementing dental quality measures include: a de-emphasis on patient-centeredness with process-based quality measures, an incentivization of unethical behavior due to fee-for-service reimbursement systems, the risk of compromising patient and provider autonomy with plan-level measures, a disproportionate benefits of dental quality measurement going toward payers, and the risk of alienating smaller dental offices due to the resource-intensive nature of quality measurement. CONCLUSION: As our medical counterparts have embraced quality measurement for improved health outcomes, so too must the dental profession. Our ultimate goal is to ensure the delivery of high quality, patient-centered dental care and effective quality measurement is the first step. By continuously monitoring the performance of dental quality measures and their continued refinement when unintended consequences are observed, we can improve patient and population health outcomes.

Do "Moderate" Drinkers Have Reduced Mortality Risk? A Systematic Review and Meta-Analysis of Alcohol Consumption and All-Cause Mortality.

OBJECTIVE: Previous meta-analyses of cohort studies indicate a J-shaped relationship between alcohol consumption and allcause mortality, with reduced risk for low-volume drinkers. However, low-volume drinkers may appear healthy only because the "abstainers" with whom they are compared are biased toward ill health. The purpose of this study was to determine whether misclassifying former and occasional drinkers as abstainers and other potentially confounding study characteristics underlie observed positive health outcomes for lowvolume drinkers in prospective studies of all-cause mortality. METHOD: A systematic review and meta-regression analysis of studies investigating alcohol use and mortality risk after controlling for quality-related study characteristics was conducted in a population of 3,998,626 individuals, among whom 367,103 deaths were recorded. RESULTS: Without adjustment, meta-analysis of all 87 included studies replicated the classic J-shaped curve, with low-volume drinkers (1.3-24.9 g ethanol per day) having reduced mortality risk (RR = 0.86, 95% CI [0.83, 0.90]). Occasional drinkers (<1.3 g per day) had similar mortality risk (RR = 0.84, 95% CI [0.79, 0.89]), and former drinkers had elevated risk (RR = 1.22, 95% CI [1.14, 1.31]). After adjustment for abstainer biases and quality-related study characteristics, no significant reduction in mortality risk was observed for low-volume drinkers (RR = 0.97, 95% CI [0.88, 1.07]). Analyses of higher-quality bias-free studies also failed to find reduced mortality risk for low-volume alcohol drinkers. Risk estimates for occasional drinkers were similar to those for low- and medium-volume drinkers. CONCLUSIONS: Estimates of mortality risk from alcohol are significantly altered by study design and characteristics. Meta-analyses adjusting for these factors find that low-volume alcohol consumption has no net mortality benefit compared with lifetime abstention or occasional drinking. These findings have implications for public policy, the formulation of low-risk drinking guidelines, and future research on alcohol and health.