Analysis of the phenotypic variability of twenty f3 biparental populations of bread wheat (Triticum aestivum L.) evaluated under semi-arid environment

This research was conducted to screen and analyse the variability within twenty F3 populations of bread wheat (Triticum aestivum L.) generated by Line x Tester mating design. The results indicated the presence of sufficient variability within and between F3 populations for the eight measured variables, which represent plant phenology, physiology, yield and yield components. Different populations have been identified to improve the measured variables separately. The number of spikes appeared to be the most important determinant of grain yield. PCA and cluster analyses indicated that the Acsad1069/El Wifak and Acsad1135/Hidhab, with a relatively high grain yield,  aboveground biomass and 1000 grains weight, are the best F3 populations to improve the productivity. However, Acsad899/Rmada and Acsad1135/Rmada populations were earlier and had a low number of spikes. These populations had also favorable genes for heat tolerance.Keywords: Triticum aestivum L.; Variability; Selection; Tolerance; Yield

Analysis of MicroRNA Expression in the Prepubertal Testis

Only thirteen microRNAs are conserved between D. melanogaster and the mouse; however, conditional loss of miRNA function through mutation of Dicer causes defects in proliferation of premeiotic germ cells in both species. This highlights the potentially important, but uncharacterized, role of miRNAs during early spermatogenesis. The goal of this study was to characterize on postnatal day 7, 10, and 14 the content and editing of murine testicular miRNAs, which predominantly arise from spermatogonia and spermatocytes, in contrast to prior descriptions of miRNAs in the adult mouse testis which largely reflects the content of spermatids. Previous studies have shown miRNAs to be abundant in the mouse testis by postnatal day 14; however, through Next Generation Sequencing of testes from a B6;129 background we found abundant earlier expression of miRNAs and describe shifts in the miRNA signature during this period. We detected robust expression of miRNAs encoded on the X chromosome in postnatal day 14 testes, consistent with prior studies showing their resistance to meiotic sex chromosome inactivation. Unexpectedly, we also found a similar positional enrichment for most miRNAs on chromosome 2 at postnatal day 14 and for those on chromosome 12 at postnatal day 7. We quantified in vivo developmental changes in three types of miRNA variation including 5′ heterogeneity, editing, and 3′ nucleotide addition. We identified eleven putative novel pubertal testis miRNAs whose developmental expression suggests a possible role in early male germ cell development. These studies provide a foundation for interpretation of miRNA changes associated with testicular pathology and identification of novel components of the miRNA editing machinery in the testis

Mutations in KEOPS-Complex Genes Cause Nephrotic Syndrome with Primary Microcephaly

Galloway-Mowat syndrome (GAMOS) is an autosomal-recessive disease characterized by the combination of early-onset nephrotic syndrome (SRNS) and microcephaly with brain anomalies. Here we identified recessive mutations in OSGEP, TP53RK, TPRKB, and LAGE3, genes encoding the four subunits of the KEOPS complex, in 37 individuals from 32 families with GAMOS. CRISPR-Cas9 knockout in zebrafish and mice recapitulated the human phenotype of primary microcephaly and resulted in early lethality. Knockdown of OSGEP, TP53RK, or TPRKB inhibited cell proliferation, which human mutations did not rescue. Furthermore, knockdown of these genes impaired protein translation, caused endoplasmic reticulum stress, activated DNA-damage-response signaling, and ultimately induced apoptosis. Knockdown of OSGEP or TP53RK induced defects in the actin cytoskeleton and decreased the migration rate of human podocytes, an established intermediate phenotype of SRNS. We thus identified four new monogenic causes of GAMOS, describe a link between KEOPS function and human disease, and delineate potential pathogenic mechanisms

Comparison of Tendon Lengthening with Traditional vs. Accelerated Rehab Following Achilles Tendon Repair: A Randomized Controlled Trial

Objectives: Operative repair of Achilles tendon ruptures have shown successful outcomes. However, little is know about the amount of tendon or repair site lengthening after repair and if lengthening is affected by rehab protocols. The purpose of our study was to compare lengthening of the Achilles tendon after surgical repair, comparing traditional and accelerated rehab protocols. Methods: Twenty patients undergoing primary repair of Achilles tendon ruptures were assessed for participation. We performed a prospective randomized controlled trial in accordance with the CONSORT (Consolidated Standards of Reporting Trials) 2010 statement. The study arms included operative repair of Achilles tendon rupture with either accelerated (graduated weight bearing at 2 weeks) or traditional rehab (weight bearing at 6 weeks). During repair, two 2-mm tantalum beads with laser-etched holes were sutured to the Achilles tendon at the repair site. Beads were evaluated via CT scans immediately post-operatively and at 12 weeks. X-rays were obtained at time 0 and then at 2, 6, and 12 weeks. The primary outcome of the study was the difference in tendon or repair site lengthening between the study arms. Randomization was by a computerized algorithm. The observer was blinded and the patient was not blinded to the intervention. Results: Zero patients declined participation and all 20 patients were included for final analysis. All patients showed statistically significant lengthening at two weeks following surgery. There was a trend toward increased lengthening at 6 weeks in the accelerated rehab group (9.9mm, range 2.6 -13.9mm) compared to the traditional rehab group (4.1mm, range 1.5 -9.0mm), although this was not statistically significant; p = .07. However the final amount of tendon lengthening at 12 weeks after surgery was not different between the accelerated rehab group (14.4mm, range 11.7 -17.0mm) and the traditional rehab group (13.4mm, range 10.7 -17.0mm); p = .38. Conclusion: This study\u27s findings suggest that all patients undergoing operative repair of Achilles tendon ruptures have significant lengthening after surgery. Although there was a trend toward increased lengthening at 6 weeks in the accelerated rehab group, there was no difference in tendon lengthening at final follow up between the two groups

Addition of iptacopan, an oral factor B inhibitor, to eculizumab in patients with paroxysmal nocturnal haemoglobinuria and active haemolysis: an open-label, single-arm, phase 2, proof-of-concept trial

Background: The haematological benefit of standard-of-care anti-C5 treatment for haemolytic paroxysmal nocturnal haemoglobinuria is limited by residual intravascular haemolysis or emerging C3-mediated extravascular haemolysis. Therefore, the aim of this phase 2 study was to assess the safety, tolerability, pharmacokinetics and pharmacodynamics, and activity of the new complement factor B inhibitor, iptacopan, in patients with paroxysmal nocturnal haemoglobinuria who have active haemolysis despite anti-C5 therapy. Methods: In this multicentre, open-label, single-arm, phase 2 trial, we enrolled adult patients (aged 18–80 years) with paroxysmal nocturnal haemoglobinuria who showed signs of active haemolysis despite receiving eculizumab treatment. Patients were enrolled at Federico II University Hospital (Naples, Italy), Hôpital Saint-Louis (Paris, France), and University Hospital Essen (Essen, Germany). For enrolment, patients were required to show lactate dehydrogenase more than 1·5-times the upper limit of normal and a paroxysmal nocturnal haemoglobinuria type 3 erythrocyte or granulocyte clone size of 10% or greater. Patients with bone marrow failure, on systemic steroid or immunosuppressive drugs, or with severe comorbidities were excluded from the study. Iptacopan was given orally as an add-on therapy at a dose of 200 mg twice daily. The primary endpoint was the effect of iptacopan on the reduction of chronic residual intravascular haemolysis measured as change in lactate dehydrogenase from baseline value to week 13. At 13 weeks, patients could opt into a long-term study extension (ongoing), allowing for modifications of standard treatment. This trial is registered at ClinicialTrials.gov, NCT03439839. Findings: Between May 31, 2018, and April 9, 2019, ten patients had twice daily 200 mg iptacopan. Iptacopan resulted in marked reduction of lactate dehydrogenase from baseline versus at week 13 (mean 539 IU/L [SD 263] vs 235 IU/L [44], change from baseline –309·2 IU/L [SD 265·5], 90% CI –473·77 to –144·68, p=0·0081), associated with significant improvement of haemoglobin concentrations (mean 97·7 g/L [SD 10·5] vs 129·5 g/L [18·3] change from baseline 31·9 g/L [14·5], 90% CI 23·42–40·28, p<0·0001). All biomarkers of haemolysis improved on iptacopan treatment. Observed haematological benefits were maintained longer than the 13-week study period, throughout the study extension, including seven patients who stopped concomitant standard-of-care treatment and continued iptacopan as monotherapy. There were no deaths or treatment-related serious adverse events during the study period. Of three non-related serious adverse events, two occurred in the same patient (one during run-in and before exposure to iptacopan). Interpretation: Iptacopan at a chronic dose of 200 mg twice daily was well tolerated without any major drug-related safety findings and shows lactate dehydrogenase reduction and haemoglobin normalisation in most patients with paroxysmal nocturnal haemoglobinuria at week 13 and beyond, even in monotherapy. On the basis of these data, iptacopan will be tested as monotherapy in pivotal trials investigating its haematological benefit in a broader paroxysmal nocturnal haemoglobinuria population. Funding: Novartis Institutes for Biomedical Research