Prospects of endosperm DNA in maize seed characterization

DNA based characterisation of maize germplasm has become the easiest and fastest approach to identify genetic diversity as compared to phenotyping. The conventional DNA source for genotyping is the leaf which required at least 2 weeks waiting period from seed planting to leaves sampling. This work exploits the use of endosperm DNA (EDNA) for the genotyping of maize germplasm. Maize endosperm was excised from maize seeds using pli¬ers, ground and used for Genomic DNA extraction (gDNA). Leaves DNA (LDNA) was also extracted concurrently. The extracted LDNA and EDNA were quantified and subjected to SSR-PCR. The mean concentrations of DNA extracted were 1575 ng/ul for the leaves and 526 ng/ul for endosperm. Though the difference in quantity of EDNA and LDNA were highly significant, the quality (A260/A280) for both EDNA and LDNA fall within 1.6-1.8 range of pure DNA index. SSR-PCR products using phi032 were similar for both EDNA and LDNA, indicating the usability of EDNA in genotyping. This seed based method of gDNA extraction takes less than 24 hours from sampling to quantification and genotyping. It also allows germination of sampled seeds, selection before planting, avoids the delay of planting and waiting in leaf sampling and saves field space

Genetics of carotenoids for provitamin A biofortification in tropical-adapted maize

Yellow maize contains high levels of β-carotene (βC), making it an important crop for combating vitamin A deficiency through biofortification. In this study, nine maize inbred lines were selected at random from 31 provitamin A (PVA) maize inbred lines and crossed in a partial diallel mating design to develop 36 crosses. The crosses were evaluated in the field in two locations (Samaru and Kerawa) and their seed carotenoid content were determined by high-performance liquid chromatography. The modes of gene action, heritability, and correlations between agronomic traits and carotenoid content were estimated. Additive genetic variances (σ2a) were lower than non-additive genetic variances (σ2d) for all the carotenoids, plant height (PH), and grain yield (GY), suggesting a preponderance of non-additive gene action. Broad-sense heritability (H2) was high (H2 > 60%) for zeaxanthin, days to anthesis, and PH, moderate (30% < H2 < 60%) for lutein and GY, and low (H2 < 30%) for alpha carotene, beta cryptoxanthin, βC, and PVA. Genetic advance as a percentage of mean, considered with H2, also suggests a preponderance of non-additive gene action for PVA carotenoids. Hybrid variety development is thus an appropriate approach to improving grain yield and PVA. GY showed no significant genotypic correlations with carotenoid content, suggesting that these traits can be improved concurrently. Thus, there is ample scope for improvement of PVA and GY in the sample of tropical-adapted maize

Randomized controlled trial of fixed low-vs moderate-dose hydroxyurea for primary stroke prevention in Sub-Saharan Africa: Final results of the Spring Trial

Introduction: In children with sickle cell anemia (SCA) without transcranial Doppler (TCD) screening, the incidence rates of ischemic strokes is approximately the same among children living in low- and high- low-resource settings (Pediatr Neurol. 2019;95:73-78.) with a prevalence of ~ 11%. However, in high-income settings, the standard use of TCD ultrasonography, coupled initially with monthly blood transfusion therapy has dropped the stroke prevalence to &lt; 1%. In a low-income setting, such as Nigeria, where 50% of children in the world with SCA are born (150,000 per year), initial monthly blood transfusion therapy is not practical for most children.In the Stroke Prevention in Nigeria (SPIN) Feasibility Trial (NCT01801423), fixed moderate-dose hydroxyurea was associated with a decreased rate of strokes in children with SCA and abnormal time-averaged mean of the maximum velocity (TAMMV) TCD measurements (≥200cm/sec) when compared to no treatment in the STOP Trial, 0.76 and 10.7 strokes per 100 person-years, repsectively (Am J Hematol. 2020). Based on the success of the SPIN trial, plus the challenges of real-world implementation of a government-supported primary stroke prevention programs for estimated 40,0000 children with SCA in three states in Nigeria, we tested the hypothesis that fixed-moderate dose (~20 mg/kg/day) hydroxyurea therapy for primary stroke prevention results in a 66% relative risk reduction (9 to 3 events per 100 person-years) when compared to fixed low-dose hydroxyurea (~10 mg/kg/day) therapy in a randomized controlled trial (The SPRING Trial; NCT02560935).Methods: In this partial-blind controlled phase III trial, we randomly assigned children between 5 and 12 years of age with SCA and a TCD time-averaged mean of the maximum velocity (TAMMV) ≥ 200 cm/sec measured independently twice or TAMMV ≥220 cm/sec once at study screening to receive fixed low-dose or fixed moderate-dose hydroxyurea. The primary endpoint was a clinical stroke or a transient ischemic attack (TIA). Myelosuppression was assessed with monthly complete blood counts (CBCs). Adherence to hydroxyurea was primarily based on an increase in MCV from baseline and monthly pill count return as a percent of dispensed pills. Hemoglobin F levels were measured at baseline, annually and upon trial exit. To evaluate the safety of hydroxyurea in the trial, children attending the same SCA clinics with TCD (TAMMV) &lt;200 cm/sec at study screening were prospectively followed with biweekly phone calls and annual research visits.Results: A total of 220 children (mean age: 7.5 years, 51.8% female) were randomly assigned to fixed low- (10 mg/kg/day) or moderate- (20 mg/kg/day) dose hydroxyurea, and were followed for a median of 2.4 years (IQR 2.0-2.8). NINDS Clinical Trials leaders stopped the trial early because of futility for the primary outcome. In the fixed low- and moderate-dose hydroxyurea groups, the incidence rates of strokes per 100 person-years were 1.19 and 1.92 respectively, with an incidence rate ratio of 1.60 (95% CI: 0.31-10.34), p = 0.768. The incidence rate ratio of mortality when comparing the children treated with low- and moderate- fixed-dose hydroxyurea to the non-elevated TCD group (no hydroxyurea therapy, n= 211) was 1.97 (95% CI: 0.64-6.02) and 0.47 (95% CI: 0.05-2.38), p = 0.265 and 0.545, respectively. Returned pills during the trial was 5.4% and 4.8% in the fixed low- and moderate-dose groups, respectively, p= 0.144. MCV from baseline to endpoint increased 1.5fl and 7.2 fl in the fixed low- and moderate-dose groups, respectively, p&lt;0.001. Upon exit from the trial 29.4% and 66.7% of the fixed- low and moderate -dose groups, respectively, had either hemoglobin level ≥ 9.0 g/dl, or a fetal hemoglobin level ≥ 20%.Conclusions: For primary stroke prevention in children with SCA, fixed low-dose, when compared to fixed moderate-dose hydroxyurea therapy, demonstrated no difference in the incidence rate of strokes. Both fixed low- and moderate -dose hydroxyurea doses are superior to no treatment for primary stroke prevention with abnormal TCD values. In partnership with Katsina, Kano, and Kaduna health department's leaders in Nigeria, 9 distinct SCA and primary stroke prevention clinics have been established, with the provision of free fixed low-dose hydroxyurea therapy (Bond Chemical, Nigeria; $0.15 per 500 mg) for abnormal TCD values, and biannual CBCs as standard care ,for over 40,000 children with SCA