Assessment of genetic diversity among 16 promising cultivars of ginger using cytological and molecular markers

Ginger (Zingiber officinale Roscoe) is an economically important plant, valued all over the world. The existing variation among 16 promising cultivars as observed through differential rhizome yield (181.9 to 477.3 g) was proved to have a genetic basis using different genetic markers such as karyotype, 4C nuclear DNA content and random amplified polymorphic DNA (RAPD). The karyotypic analysis revealed a differential distribution of A, B, C, D and E type of chromosomes among different cultivars as represented by different karyotype formulas. A significant variation of 4C DNA content was recorded in ginger at an intraspecific level with values ranging from 17.1 to 24.3 pg. RAPD analysis revealed a differential polymorphism of DNA showing a number of polymorphic bands ranging from 26 to 70 among 16 cultivars. The RAPD primers OPC02, OPA02, OPD20 and OPN06 showing strong resolving power were able to distinguish all 16 cultivars. The extent of genetic diversity among these cultivars was computed through parameters of gene diversity, sum of allele numbers per locus and Shannon's information indices. Cluster analysis, Nei's genetic similarity and genetic distances, distribution of cultivars into special distance classes and principal coordinate analysis and the analysis of molecular variance suggested a conspicuous genetic diversity among different cultivars studied. The genetic variation thus detected among promising cultivars of ginger has significance for ginger improvement programs. Key words: Ginger, Karyotype, RAPD Introduction Zingiber officinale is rich in secondary metabolites such as oleoresin. It possesses an unique combination of properties like anti-inflammatory, aphrodisiac, antioxidant and antibacterial activity S. Nayak et al. · Genetic Diversity among Ginger Cultivars In the past decade, DNA polymorphism has become the marker of choice for the identification and characterization of plants. It is a relatively reliable, generally applicable method to obtain large samples of markers from any species of plant. However, each marker system samples a different fraction of the genomes and therefore has a different resolving power, range of applicability and probability of homology. The random amplified polymorphic DNA (RAPD) technique has been widely used in cultivar identification programs The objectives of this study were to (1) fingerprint ginger cultivars for identification and (2) detect the genetic diversity and relatedness of 16 cultivars sampled from different geographical regions using karyotypic analysis, 4C DNA content and RAPD analysis. In this study, many analytical procedures such as a n-j method, bootstrapping, spatial genetic structure analysis (SGS), and analysis of molecular variance (AMOVA) have been widely used to derive genetic distances among cultivars and to assess the structure of genetic data in a reduced dimensional space. Materials and Methods Plant materials Sixteen promising cultivars of ginger (Zingiber officinale) were included in the present study which were collected from the turmeric germplasm collection of the Orissa University of Agriculture and Techology (OUAT), Bhubaneswar, Orissa, India. These cultivars were initially collected from different parts of India Karyotype analysis Young and healthy root-tips of different cultivars of ginger were pre-treated in a (0.02 m) hydroxyquinoline mixture (1:1) for 3.5 h at 14 ∞C followed by overnight fixation in propionic ethanol. Chromosome staining was made in 2% lacto propionic orcin after cold hydrolysis in 5 n HCl for 7 min. Root-tips were squashed in 45% propionic acid. Ten well scattered metaphase plates were selected for karyotype analysis of each species. The chromosome morphology was determined following the method of 4C DNA content For Feulgen cytophotometric estimation of 4C DNA, ten fixed root-tips from each cultivar (2n = 22 chromosomes) were hydrolysed in 1 n HCl for 12 min at 60 ∞C, washed in distilled water and stained in Schiff's reagent for 2 h at 14 ∞C; each root-tip squash was prepared in 75% acetic acid. Ten scorings were made from each slide and the 4C DNA content was estimated from metaphase chromosomes using a NIKON Optiphot microscope with a microspectrophotometer following the method of Sharma and Sharma (1980) with monochromatic light at 550 nm. In situ DNA values were obtained on the basis of optical density measurements which were converted to picograms (pg) using Vant Hoff's 4C nuclear DNA value (67.1 pg) for Allium cepa as standard Isolation of DNA Total plant DNA was isolated from fresh and young leaves. The leaves were harvested freshly and washed thoroughly with cold autoclaved distilled water and then blotted to dry. About 2 g leaf was excised from the upper tip portion of the buds. DNA extraction was done on the day of collection. The genomic DNA was isolated following the protocol of Doyle and Doyle (1990) with a little modification. Insoluble poly(vinylpyrrolidone) was added to the leaf tissue prior to grinding. The crude DNA was purified with RNase A (@ 60 µg ml Ð1 of DNA solution) followed by washing with S. Nayak et al. · Genetic Diversity among Ginger Cultivars 487 purified chloroform/isoamylalcohol (24:1). To test the quality and quantity of the purified DNA, the samples were electrophoresed in a 0.8% agarose gel along with a known amount of uncut lambda DNA (Bangalore Genei Pvt. Ltd, Bangalore, India) as standard. The sample DNA was diluted as 25 ng µl Ð1 for RAPD-PCR analysis. RAPD amplification Twenty random decamer primers (Operon Tech., USA) from A, C, D and N series (OPA02, 03, 04, 08, 16; OPAF14; OPC02, 05; OPD03, 07, 08, 18, 20; and OPN02, 03, 04, 06, 07, 10, 12) were used for RAPD analysis. RAPD assays were performed in a final volume of 25 µl containing 10 mm Tris-HCl [tris(hydroxymethyl)aminomethane], pH 9.0, 1.5 mm MgCl 2 , 50 mm KCl and 0.01% gelatin, 200 µm of each dNTPs, 0.4 µm primer, 25 ng template DNA and 0.5 unit of Taq DNA polymerase (Bangalore Genei, Bangalore, India). The RAPD analysis was performed as per the methodology described by The amplification products were electrophoresed in 1.5% agarose gel containing ethidium bromide (@ 0.5 µg ml Ð1 ) in TAE buffer (40 mm Tris base, 20 mm sodium acetate, 20 mm EDTA, glacial acetic acid to pH 7.2) for 3 h at 60 V. A total of 2.5 µl loading buffer (1.0 X TAE, 50% glycerol, 0.25% bromophenol blue and 0.25% xylene cyanol) was added to each reaction before electrophoresis. After electrophoresis, the gels were observed under an UV-transilluminator, documented in Gel-Doc 2000 (Bio-Rad) and photographed. Resolving power According to Prevost and Wilkinson (1999) the resolving power (Rp) of a primer is: Rp = Σ IB, where IB (band informativeness) takes the value of: 1-[2 ¥ (0.5 Ð p)], p being the proportion of the 16 genotypes (ginger cultivars analyzed) containing the band. Data collection and analysi

Factors associated with stigma and manifestations experienced by Indian health care workers involved in COVID-19 management in India: A qualitative study

Healthcare personnel who deal with COVID-19 experience stigma. There is a lack of national-level representative qualitative data to study COVID-19-related stigma among healthcare workers in India. The present study explores factors associated with stigma and manifestations experienced by Indian healthcare workers involved in COVID-19 management. We conducted in-depth interviews across 10 centres in India, which were analysed using NVivo software version 12. Thematic and sentiment analysis was performed to gain deep insights into the complex phenomenon by categorising the qualitative data into meaningful and related categories. Healthcare workers (HCW) usually addressed the stigma they encountered when doing their COVID duties under the superordinate theme of stigma. Among them, 77.42% said they had been stigmatised in some way. Analyses revealed seven interrelated themes surrounding stigma among healthcare workers. It can be seen that the majority of the stigma and coping sentiments fall into the mixed category, followed by the negative sentiment category. This study contributes to our understanding of stigma and discrimination in low- and middle-income settings. Our data show that the emergence of fear of the virus has quickly turned into a stigma against healthcare workers

Repositioning of the global epicentre of non-optimal cholesterol

High blood cholesterol is typically considered a feature of wealthy western countries(1,2). However, dietary and behavioural determinants of blood cholesterol are changing rapidly throughout the world(3) and countries are using lipid-lowering medications at varying rates. These changes can have distinct effects on the levels of high-density lipoprotein (HDL) cholesterol and non-HDL cholesterol, which have different effects on human health(4,5). However, the trends of HDL and non-HDL cholesterol levels over time have not been previously reported in a global analysis. Here we pooled 1,127 population-based studies that measured blood lipids in 102.6 million individuals aged 18 years and older to estimate trends from 1980 to 2018 in mean total, non-HDL and HDL cholesterol levels for 200 countries. Globally, there was little change in total or non-HDL cholesterol from 1980 to 2018. This was a net effect of increases in low- and middle-income countries, especially in east and southeast Asia, and decreases in high-income western countries, especially those in northwestern Europe, and in central and eastern Europe. As a result, countries with the highest level of non-HDL cholesterol-which is a marker of cardiovascular riskchanged from those in western Europe such as Belgium, Finland, Greenland, Iceland, Norway, Sweden, Switzerland and Malta in 1980 to those in Asia and the Pacific, such as Tokelau, Malaysia, The Philippines and Thailand. In 2017, high non-HDL cholesterol was responsible for an estimated 3.9 million (95% credible interval 3.7 million-4.2 million) worldwide deaths, half of which occurred in east, southeast and south Asia. The global repositioning of lipid-related risk, with non-optimal cholesterol shifting from a distinct feature of high-income countries in northwestern Europe, north America and Australasia to one that affects countries in east and southeast Asia and Oceania should motivate the use of population-based policies and personal interventions to improve nutrition and enhance access to treatment throughout the world.Peer reviewe

Rising rural body-mass index is the main driver of the global obesity epidemic in adults

Body-mass index (BMI) has increased steadily in most countries in parallel with a rise in the proportion of the population who live in cities(.)(1,2) This has led to a widely reported view that urbanization is one of the most important drivers of the global rise in obesity(3-6). Here we use 2,009 population-based studies, with measurements of height and weight in more than 112 million adults, to report national, regional and global trends in mean BMI segregated by place of residence (a rural or urban area) from 1985 to 2017. We show that, contrary to the dominant paradigm, more than 55% of the global rise in mean BMI from 1985 to 2017-and more than 80% in some low- and middle-income regions-was due to increases in BMI in rural areas. This large contribution stems from the fact that, with the exception of women in sub-Saharan Africa, BMI is increasing at the same rate or faster in rural areas than in cities in low- and middle-income regions. These trends have in turn resulted in a closing-and in some countries reversal-of the gap in BMI between urban and rural areas in low- and middle-income countries, especially for women. In high-income and industrialized countries, we noted a persistently higher rural BMI, especially for women. There is an urgent need for an integrated approach to rural nutrition that enhances financial and physical access to healthy foods, to avoid replacing the rural undernutrition disadvantage in poor countries with a more general malnutrition disadvantage that entails excessive consumption of low-quality calories.Peer reviewe

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Global variations in diabetes mellitus based on fasting glucose and haemogloblin A1c

Fasting plasma glucose (FPG) and haemoglobin A1c (HbA1c) are both used to diagnose diabetes, but may identify different people as having diabetes. We used data from 117 population-based studies and quantified, in different world regions, the prevalence of diagnosed diabetes, and whether those who were previously undiagnosed and detected as having diabetes in survey screening had elevated FPG, HbA1c, or both. We developed prediction equations for estimating the probability that a person without previously diagnosed diabetes, and at a specific level of FPG, had elevated HbA1c, and vice versa. The age-standardised proportion of diabetes that was previously undiagnosed, and detected in survey screening, ranged from 30% in the high-income western region to 66% in south Asia. Among those with screen-detected diabetes with either test, the agestandardised proportion who had elevated levels of both FPG and HbA1c was 29-39% across regions; the remainder had discordant elevation of FPG or HbA1c. In most low- and middle-income regions, isolated elevated HbA1c more common than isolated elevated FPG. In these regions, the use of FPG alone may delay diabetes diagnosis and underestimate diabetes prevalence. Our prediction equations help allocate finite resources for measuring HbA1c to reduce the global gap in diabetes diagnosis and surveillance.peer-reviewe

Causes of blindness and vision impairment in 2020 and trends over 30 years, and prevalence of avoidable blindness in relation to VISION 2020: the Right to Sight: an analysis for the Global Burden of Disease Study

Background: Many causes of vision impairment can be prevented or treated. With an ageing global population, the demands for eye health services are increasing. We estimated the prevalence and relative contribution of avoidable causes of blindness and vision impairment globally from 1990 to 2020. We aimed to compare the results with the World Health Assembly Global Action Plan (WHA GAP) target of a 25% global reduction from 2010 to 2019 in avoidable vision impairment, defined as cataract and undercorrected refractive error. Methods: We did a systematic review and meta-analysis of population-based surveys of eye disease from January, 1980, to October, 2018. We fitted hierarchical models to estimate prevalence (with 95% uncertainty intervals [UIs]) of moderate and severe vision impairment (MSVI; presenting visual acuity from <6/18 to 3/60) and blindness (<3/60 or less than 10° visual field around central fixation) by cause, age, region, and year. Because of data sparsity at younger ages, our analysis focused on adults aged 50 years and older. Findings: Global crude prevalence of avoidable vision impairment and blindness in adults aged 50 years and older did not change between 2010 and 2019 (percentage change −0·2% [95% UI −1·5 to 1·0]; 2019 prevalence 9·58 cases per 1000 people [95% IU 8·51 to 10·8], 2010 prevalence 96·0 cases per 1000 people [86·0 to 107·0]). Age-standardised prevalence of avoidable blindness decreased by −15·4% [–16·8 to −14·3], while avoidable MSVI showed no change (0·5% [–0·8 to 1·6]). However, the number of cases increased for both avoidable blindness (10·8% [8·9 to 12·4]) and MSVI (31·5% [30·0 to 33·1]). The leading global causes of blindness in those aged 50 years and older in 2020 were cataract (15·2 million cases [9% IU 12·7–18·0]), followed by glaucoma (3·6 million cases [2·8–4·4]), undercorrected refractive error (2·3 million cases [1·8–2·8]), age-related macular degeneration (1·8 million cases [1·3–2·4]), and diabetic retinopathy (0·86 million cases [0·59–1·23]). Leading causes of MSVI were undercorrected refractive error (86·1 million cases [74·2–101·0]) and cataract (78·8 million cases [67·2–91·4]). Interpretation: Results suggest eye care services contributed to the observed reduction of age-standardised rates of avoidable blindness but not of MSVI, and that the target in an ageing global population was not reached. Funding: Brien Holden Vision Institute, Fondation Théa, The Fred Hollows Foundation, Bill & Melinda Gates Foundation, Lions Clubs International Foundation, Sightsavers International, and University of Heidelberg