Mapping of the Rpv Resistance Gene against Downy Mildew in Pea (Pisum sativum L.)

Die Entwicklung von Sorten, welche eine Resistenz gegen den Erreger des Falschen Mehltaus (Peronospora viciae f. sp. pisi) aufweisen, hat für Erbsenzüchter einen hohen Stellenwert, da dieser Pilz deutliche Ertragsminderungen und Qualitätseinbußen verursachen kann. In der vorliegenden Arbeit wurde eine spaltende Nachkommenschaft von 335 F2-Nachkommen einer Initialkreuzung zwischen der anfälligen Hochleistunssorte `Topaz´ und der resistenten Zuchtlinie `Gen. 27´ erstellt. Jeweils 10 F3-Nachkommen dieser 335 F2-Pflanzen wurden in der Klimakammer auf Resistenz gegen den Falschen Mehltau getestet um auf den Genotyp der ursprünglichen F2-Pflanze schließen zu können. Das Verhältnis von 94 homozygot resistenten, 161 heterozygot resistenten und 80 homozygot anfälligen F2-Pflanzen weicht nicht signifikant von einer 1:2:1-Spaltung ab, die für das Vorliegen eines dominanten Resistenzgens erwartet wird. Das Resis­tenzgen Rpv wurde auf Kopplungsgruppe 1 der Erbse kartiert. Diese Kopplungsgruppe wurde anschließend mit weiteren molekularen Markern, die in der Literatur beschrieben wurden, gesättigt. Der Marker AD147 wurde dabei als proximaler Marker mit der geringsten Distanz (4,4 cM) bestimmt, während in distaler Position AB28 eine Distanz von 18,8 cM aufwies. Durch Nutzung der Synthenie zum Medicago truncatula Chromosom 5 konnten keine enger gekoppelten Marker ermittelt werden. Der Nutzen der identifizierten Marker in unabhängigem Zuchtmaterial konnte jedoch demonstriert und auch die Basis für eine zukünftige Feinkartierung des Resis­tenzgens gelegt werden.Downy mildew causes severe yield and quality losses in pea (Pisum sativum L.). Therefore, the development of downy mildew resistant varieties is of high priority for pea breeders. Within this study in total 335 F3 families from a cross of the highly susceptible green pea variety `Topaz´ with the resistant breeding line `Gen. 27´ were tested for resistance behaviour against Peronospora viciae f. sp. pisi to determine the genotype of the corresponding F2 parental plants. The ratio 94:161:80 for homozygous resistant, heterozygous resistant and homozygous susceptible F2 plants was not significantly different from 1:2:1, expected for the effect of a single dominant resistance gene. The resistance gene Rpv was mapped to linkage group 1 of the pea genetic map. This linkage group was saturated by molecular markers available from lite­rature. The marker AD147 was identified as nearest proxi­mal flanking marker with 4.4 cM distance, and in distal position marker AB28 with 18.8 cM distance. Further marker saturation using the syntenic relationship of P. sativum and Medicago truncatula was not effective. The usefulness of the identified markers for marker assisted selection has been confirmed in independent pea breeding material and the results of this study should lay the basis for future fine mapping studies

Toward improving practices for submission of diagnostic tissue blocks for National Cancer Institute clinical trials

OBJECTIVES: The National Cancer Institute (NCI) National Clinical Trials Network performs phase II and III clinical trials, which increasingly rely on the submission of diagnostic formalin-fixed, paraffin-embedded tissue blocks for biomarker assessment. Simultaneously, advances in precision oncology require that clinical centers maintain diagnostic specimens for ancillary, standard-of-care diagnostics. This has caused tissue blocks to become a limited resource for advancing the NCI clinical trial enterprise and the practice of modern molecular pathology. METHODS: The NCI convened a 1-day workshop of multidisciplined experts to discuss barriers and strategic solutions to facilitate diagnostic block submission for clinical trial science, from the perspective of patient advocates, legal experts, pathologists, and clinical oncologists. RESULTS: The expert views and opinions were carefully noted and reported. CONCLUSIONS: Recommendations were proposed to reduce institutional barriers and to assist organizations in developing clear policies regarding diagnostic block submission for clinical trials

Altered engagement of the speech motor network is associated with reduced phonological working memory in autism

Nonword repetition, a common clinical measure of phonological working memory, involves component processes of speech perception, working memory, and speech production. Autistic children often show behavioral challenges in nonword repetition, as do many individuals with communication disorders. It is unknown which subprocesses of phonological working memory are vulnerable in autistic individuals, and whether the same brain processes underlie the transdiagnostic difficulty with nonword repetition. We used functional magnetic resonance imaging (fMRI) to investigate the brain bases for nonword repetition challenges in autism. We compared activation during nonword repetition in functional brain networks subserving speech perception, working memory, and speech production between neurotypical and autistic children. Autistic children performed worse than neurotypical children on nonword repetition and had reduced activation in response to increasing phonological working memory load in the supplementary motor area. Multivoxel pattern analysis within the speech production network classified shorter vs longer nonword-repetition trials less accurately for autistic than neurotypical children. These speech production motor-specific differences were not observed in a group of children with reading disability who had similarly reduced nonword repetition behavior. These findings suggest that atypical function in speech production brain regions may contribute to nonword repetition difficulties in autism.R01 DC011339 - NIDCD NIH HHS; R21 DC017576 - NIDCD NIH HHS; R03 DC014045 - NIDCD NIH HHS; T32 DC000038 - NIDCD NIH HHSPublished versio

New Synthetic Thrombin Inhibitors: Molecular Design and Experimental Verification

BACKGROUND: The development of new anticoagulants is an important goal for the improvement of thromboses treatments. OBJECTIVES: The design, synthesis and experimental testing of new safe and effective small molecule direct thrombin inhibitors for intravenous administration. METHODS: Computer-aided molecular design of new thrombin inhibitors was performed using our original docking program SOL, which is based on the genetic algorithm of global energy minimization in the framework of a Merck Molecular Force Field. This program takes into account the effects of solvent. The designed molecules with the best scoring functions (calculated binding energies) were synthesized and their thrombin inhibitory activity evaluated experimentally in vitro using a chromogenic substrate in a buffer system and using a thrombin generation test in isolated plasma and in vivo using the newly developed model of hemodilution-induced hypercoagulation in rats. The acute toxicities of the most promising new thrombin inhibitors were evaluated in mice, and their stabilities in aqueous solutions were measured. RESULTS: New compounds that are both effective direct thrombin inhibitors (the best K(I) was <1 nM) and strong anticoagulants in plasma (an IC(50) in the thrombin generation assay of approximately 100 nM) were discovered. These compounds contain one of the following new residues as the basic fragment: isothiuronium, 4-aminopyridinium, or 2-aminothiazolinium. LD(50) values for the best new inhibitors ranged from 166.7 to >1111.1 mg/kg. A plasma-substituting solution supplemented with one of the new inhibitors prevented hypercoagulation in the rat model of hemodilution-induced hypercoagulation. Activities of the best new inhibitors in physiological saline (1 µM solutions) were stable after sterilization by autoclaving, and the inhibitors remained stable at long-term storage over more than 1.5 years at room temperature and at 4°C. CONCLUSIONS: The high efficacy, stability and low acute toxicity reveal that the inhibitors that were developed may be promising for potential medical applications

National Scale Real-Time Surveillance of SARS-CoV-2 Variants Dynamics by Wastewater Monitoring in Israel

In this report, we describe a national-scale monitoring of the SARS-CoV-2 (SC-2) variant dynamics in Israel, using multiple-time sampling of 13 wastewater treatment plants. We used a combination of inclusive and selective quantitative PCR assays that specifically identify variants A19/A20 or B.1.1.7 and tested each sample for the presence and relative viral RNA load of each variant. We show that between December 2020 and March 2021, a complete shift in the SC-2 variant circulation was observed, where the B.1.1.7 replaced the A19 in all examined test points. We further show that the normalized viral load (NVL) values and the average new cases per week reached a peak in January 2021 and then decreased gradually in almost all test points, in parallel with the progression of the national vaccination campaign, during February&ndash;March 2021. This study demonstrates the importance of monitoring SC-2 variant by using a combination of inclusive and selective PCR tests on a national scale through wastewater sampling, which is far more amendable for high-throughput monitoring compared with sequencing. This approach may be useful for real-time dynamics surveillance of current and future variants, such as the Omicron (BA.1, BA.2) and other variants

Molecular Features of Cancers Exhibiting Exceptional Responses to Treatment

A small fraction of cancer patients with advanced disease survive significantly longer than patients with clinically comparable tumors. Molecular mechanisms for exceptional responses to therapy have been identified by genomic analysis of tumor biopsies from individual patients. Here, we analyzed tumor biopsies from an unbiased cohort of 111 exceptional responder patients using multiple platforms to profile genetic and epigenetic aberrations as well as the tumor microenvironment. Integrative analysis uncovered plausible mechanisms for the therapeutic response in nearly a quarter of the patients. The mechanisms were assigned to four broad categories—DNA damage response, intracellular signaling, immune engagement, and genetic alterations characteristic of favorable prognosis—with many tumors falling into multiple categories. These analyses revealed synthetic lethal relationships that may be exploited therapeutically and rare genetic lesions that favor therapeutic success, while also providing a wealth of testable hypotheses regarding oncogenic mechanisms that may influence the response to cancer therapy.[Display omitted]•Genomics of 110 patients with exceptional response to therapy profiled•Plausible molecular mechanisms related to therapy identified in ∼23% of cases•Proposed mechanisms involve DNA damage, signaling, and the immune response•Synthetic lethality with temozolomide in tumors with a defective DNA damage responseProfiling multi-platform genomics of 110 cancer patients with an exceptional therapeutic response, Wheeler et al. identify putative molecular mechanisms explaining this survival phenotype in ∼23% of cases. Therapeutic success is related to rare molecular features of responding tumors, exploiting synthetic lethality and oncogene addiction