Functional analysis of germline <em>VANGL2</em> variants using rescue assays of <em>vangl2</em> knockout zebrafish

\ua9 The Author(s) 2023. Published by Oxford University Press. Developmental studies have shown that the evolutionarily conserved Wnt Planar Cell Polarity (PCP) pathway is essential for the development of a diverse range of tissues and organs including the brain, spinal cord, heart and sensory organs, as well as establishment of the left-right body axis. Germline mutations in the highly conserved PCP gene VANGL2 in humans have only been associated with central nervous system malformations, and functional testing to understand variant impact has not been performed. Here we report three new families with missense variants in VANGL2 associated with heterotaxy and congenital heart disease p.(Arg169His), non-syndromic hearing loss p.(Glu465Ala) and congenital heart disease with brain defects p.(Arg135Trp). To test the in vivo impact of these and previously described variants, we have established clinically-relevant assays using mRNA rescue of the vangl2 mutant zebrafish. We show that all variants disrupt Vangl2 function, although to different extents and depending on the developmental process. We also begin to identify that different VANGL2 missense variants may be haploinsufficient and discuss evidence in support of pathogenicity. Together, this study demonstrates that zebrafish present a suitable pipeline to investigate variants of unknown significance and suggests new avenues for investigation of the different developmental contexts of VANGL2 function that are clinically meaningful

P-solubilizing Bacteria as a Panacea to Alleviate Stress Effects of High Soil CaCO3 content in Phaseolus Vulgaris with Special Reference to P-Releasing Enzymes

Abstract Purpose: The present study examines the role of leguminous compost (LC), humic acid (HA), and phosphate-solubilizing bacteria (P-SB) in alleviating the stress effects of high soil CaCO3 content in Phaseolus vulgaris. Methods: Two pot trials for two consecutive seasons; fall 2019 and summer 2020 were implemented in an open greenhouse. With four replicates specified for each of five treatments, a randomized complete plot design was assigned to each trial. Results: Inoculation of calcareous soil with P-SB (a 1: 1 mixture of two Pseudomonas sp.; Ps. mallei and Ps. cepaceae) significantly exceeded LC, HA, or even LC+HA for the positive results obtained. P-SB facilitated nutrient solubility (e.g., N, K, Fe, and Mn), including conversion of insoluble phosphorous into a form available in the tested soil due to increased soil enzymatic activities (e.g., phosphatases and phytases). This mechanism, combined with a decrease in soil calcium carbonate content and an increase in cation exchange capacity (CEC) and organic matter (OM) content, increased the availability of various nutrients to plants, including P, in the soil, which contributed to the increased plant output. Adequate P content in plants led to a marked decrease in plant acid phosphatase activity under high content of CaCO3. Conclusions The study concluded that the use of P-SB promotes biological activities, nutrient availability, and thus the productivity of calcareous soils, enabling Phaseolus vulgaris plants to withstand stress produced by high CaCO3 content through the development and/or adoption of potentially effective mechanisms.</jats:p

Glucosinolates With Their Hydrolysis Products from Two Cruciferous Plants with Study of Antidiabetic Activity Based on Molecular Docking

Abstract The glucosinolates (Gls.) are natural bioactive compounds which lead to the formation of different metabolites called isothiocyanates(ITC) having various therapeutic effects. So, this study aim to isolate the glucosinolates of both Carrichtera annua L.(DC) (CA) and Farsetia aegyptia Turra (FA) belonging to Crucifereae family. The total Gls. were isolated from the aqueous methanolic extract of both plants and further purified on acidic aluminum oxide column. Some of the obtained Gls. was identified as it is using different spectroscopic measurements (UV, NMR and MS) and the rest were hydrolyzed using myrosinase enzyme to the corresponding isothiocyanates (ITC) which were identified by GC/MS. only one glucosinolate was identified in CA as: 4-methylthio-3-butenyl Gls ( MTBG). through chromatographic and spectroscopic measurements in addition to 6-methyl sulfonylhexyl isothiocyanates(ITC), while 6-methyl sulfonyl-6-hydroxy hexyl ITC, 4-pentenyl ITC, 3-Methylthio propyl ITC, 5-hydroxy pentyl ITC and 4-methylsulphinyl butyl ITC from FA. The docking study targeted a α-glucosidase and amylase, to examine a mode of action of the 4-methylthio-3-butenylglucosinolate. Molecular docking was performed to identify potency of Gls. against hyperglycemia. The data obtained revealed that the Gls. has high binding activity Via α-glucosidase and amylase. Furthermore, further Drug studies as likeness and ADME/T were performed, which proposed that their ligands may be have a good pharmacokinetic character, with no carcinogenesis effect.</jats:p

Phosphate-Solubilizing Bacteria as a Panacea to Alleviate Stress Effects of High Soil CaCO3 Content in Phaseolus vulgaris with Special Reference to P-Releasing Enzymes

The present study examines the role of leguminous compost (LC), humic acids (HA), and phosphate-solubilizing bacteria (P-SB) in alleviating the stress effects of high soil CaCO3 content in Phaseolus vulgaris. Two pot trials for two consecutive seasons; fall 2019 and summer 2020 were implemented in an open greenhouse. A mixed three-way ANOVA, two independent factors (season and soil treatments) and one within factors (time) were used with four replicates. Residual maximum likelihood (REML) analysis was used for the mixed model of the studied traits. Inoculation of calcareous soil with P-SB (a 1:1 mixture of two Pseudomonas sp.; Ps. mallei and Ps. cepaceae) significantly exceeded LC, HA, or even LC+HA for the positive results obtained. P-SB facilitated nutrient solubility (e.g., N, K, Fe, and Mn), including conversion of insoluble phosphorous into a form available in the tested soil due to increased soil enzymatic activities (e.g., phosphatases and phytases). This mechanism, combined with a decrease in soil calcium carbonate content and an increase in cation exchange capacity (CEC) and organic matter (OM) content, increased the availability of various nutrients to plants, including P, in the soil, which contributed to the increased plant output. Adequate P content in plants led to a marked decrease in plant acid phosphatase activity under high content of CaCO3. The study concluded that the use of P-SB promotes biological activities, nutrient availability, and thus the productivity of calcareous soils, enabling Phaseolus vulgaris plants to withstand stress produced by high CaCO3 content through the development and/or adoption of potentially effective mechanisms. Strong highly significant interactions between the treatments and time were observed using the Wald’s statistics test, which indicates a positive correlation.</jats:p

Phosphate-Solubilizing Bacteria as a Panacea to Alleviate Stress Effects of High Soil CaCO3 Content in Phaseolus vulgaris with Special Reference to P-Releasing Enzymes

The present study examines the role of leguminous compost (LC), humic acids (HA), and phosphate-solubilizing bacteria (P-SB) in alleviating the stress effects of high soil CaCO3 content in Phaseolus vulgaris. Two pot trials for two consecutive seasons; fall 2019 and summer 2020 were implemented in an open greenhouse. A mixed three-way ANOVA, two independent factors (season and soil treatments) and one within factors (time) were used with four replicates. Residual maximum likelihood (REML) analysis was used for the mixed model of the studied traits. Inoculation of calcareous soil with P-SB (a 1:1 mixture of two Pseudomonas sp.; Ps. mallei and Ps. cepaceae) significantly exceeded LC, HA, or even LC+HA for the positive results obtained. P-SB facilitated nutrient solubility (e.g., N, K, Fe, and Mn), including conversion of insoluble phosphorous into a form available in the tested soil due to increased soil enzymatic activities (e.g., phosphatases and phytases). This mechanism, combined with a decrease in soil calcium carbonate content and an increase in cation exchange capacity (CEC) and organic matter (OM) content, increased the availability of various nutrients to plants, including P, in the soil, which contributed to the increased plant output. Adequate P content in plants led to a marked decrease in plant acid phosphatase activity under high content of CaCO3. The study concluded that the use of P-SB promotes biological activities, nutrient availability, and thus the productivity of calcareous soils, enabling Phaseolus vulgaris plants to withstand stress produced by high CaCO3 content through the development and/or adoption of potentially effective mechanisms. Strong highly significant interactions between the treatments and time were observed using the Wald’s statistics test, which indicates a positive correlation