Targeted editing of tomato carotenoid isomerase reveals the role of 5' UTR region in gene expression regulation.

KEY MESSAGE: A deletion created by CRISPR/Cas9 system in the 5' UTR of the carotenoid isomerase gene in tomato leads to downregulation of the gene resulting in the low conversion of prolycopene to lycopene. CRISPR/Cas9 based genome editing is an effective and useful tool adopted from the bacterial immune response system for altering specific, pre-determined DNA sequences in eukaryotes. Such targeted changes are finding wide application in human health as well as in precision breeding of crop plants for improved traits. Mutations in the coding and regulatory regions can have varying impacts on the function of the gene. In the current study, we demonstrate this on tomato carotenoid isomerase, a key gene in the carotenoid biosynthesis pathway. Mutations were generated in the 5' UTR and exon 1 of the carotenoid isomerase gene using CRISPR/Cas9 expression via Agrobacterium-mediated transformation of tomato variety Periyakulam 1 (PKM1). Molecular and biochemical studies demonstrate that CRISPR-mediated point mutations in the exon sequence lead to complete knockout of protein function whereas deletion in 5' UTR region lowers the expression of the gene leading to changes in plant phenotype.

A three-dimensional finite element analysis to evaluate stress distribution tooth in tooth implant-supported prosthesis with variations in non-rigid connector design and location.

BACKGROUND: The physiologically mobile natural tooth and rigidly fixed dental implant causes different distribution of stress when connected in prosthesis and nonrigid connector compensates this. Understanding of biomechanical behavior is necessary for an adequate choice and construction of this type of rehabilitation. However, there has been insufficient research focusing on different location and type of the nonrigid connector related with the prognosis of both implant and the tooth. AIM OF THE STUDY: The purpose of this finite element (FE) analysis was to evaluate the stress distribution around bone, implant, and tooth in tooth implant fixed prosthesis under static load with variations in design and location of nonrigid connectors under simulated functional loads. MATERIALS AND METHOD: Three, 3-dimensional FE models connecting tooth and implant were constructed with different location and type of nonrigid connector. Simulated occlusal load was applied on the restorations and stresses developed in the supporting structures were monitored. RESULTS: The highest stresses were found around the implant in model with nonrigid connector placed between the tooth and implant and model with modified nonrigid connector. On the other hand, less stress was noted around the implant where nonrigid connector was placed between the implant and pontic. CONCLUSION: It is advisable to place the nonrigid connector between the implant and the pontic to protect the implant from torque effects in a tooth implant fixed prosthesis.

Investigation of Antidiabetic, Antihyperlipidemic, and In Vivo Antioxidant Properties of Sphaeranthus indicus Linn. in Type 1 Diabetic Rats: An Identification of Possible Biomarkers.

The present investigation was aimed to study the antidiabetic, antihyperlipidemic, and in vivo antioxidant properties of the root of Sphaeranthus indicus Linn. in streptozotocin- (STZ-) induced type 1 diabetic rats. Administration of ethanolic extract of Sphaeranthus indicus root (EESIR) 100 and 200 mg/kg to the STZ-induced diabetic rats showed significant (P < .01) reduction in blood glucose and increase in body weight compared to diabetic control rats. Both the doses of EESIR-treated diabetic rats showed significant (P < .01) alteration in elevated lipid profile levels than diabetic control rats. The EESIR treatment in diabetic rats produced significant increase in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and decrease in thiobarbituric acid reactive substances (TBARS) levels than diabetic control rats. Administration of EESIR 200 mg/kg produced significant (P < .01) higher antioxidant activity than EESIR 100 mg/kg. The high performance liquid chromatography (HPLC) analysis of EESIR revealed the presence of biomarkers gallic acid and quercetin. In conclusion, EESIR possess antidiabetic, antihyperlipidemic, and in vivo antioxidant activity in type 1 diabetic rats. Its antioxidant and lipid lowering effect will help to prevent diabetic complications, and these actions are possibly due to presence of above biomarkers.