Soybean isoflavonoid biosynthesis: constituents and circumstance at the transcriptomic and molecular levels

Isoflavonoids are specialized metabolites, almost exclusive to the legume family of plants. They are actors in symbiosis with nitrogen-fixing bacteria and in plant stress response. Isoflavonoids are noted for their human health benefits. Isoflavonoid content in legumes has proven to be a complex trait. The goal of the present research is to determine the mechanisms underlying isoflavonoid biosynthesis in soybean. The first approach was to unravel the genetic factors of isoflavonoid biosynthesis. A branch-point enzyme of the phenylpropanoid pathway, chalcone isomerase (CHI), catalyzes the reaction producing flavanones, the nucleus for many downstream metabolites such as isoflavonoids. I identified twelve soybean CHI genes, including five new members. The evolutionary history of the family shows that the enzymatic fold evolved from being catalytically inactive to being a chalcone-to-flavanone isomerase. Four CHIs in soybean were identified with the latter functionality. The CHIs showed differential temporal and spatial expression, pointing to the potential function of CHI1A in soybean seed isoflavonoid production. On the molecular level of organization, the long-postulated model of a subcellular isoflavonoid enzyme complex forming at the surface of the endoplasmic reticulum (ER) was substantiated in the present study. Here, I identified key players in the ‘metabolon’, established in planta subcellular localization, and investigated protein-protein interaction. The results suggest that isoflavone synthase (IFS), a cytochrome P450, is a nucleating metabolic center at the surface of the ER, interacting with upstream pathway enzymes. Finally, a transcriptomic study was undertaken to find genetic elements linked with isoflavonoid content variation in four soybean cultivars. The results suggest that competing branches of the phenylpropanoid pathway are combinatorially regulated to coordinate flux into isoflavonoid biosynthesis. The candidate genes encode enzymes in the overlapping pathways, several transcription factors, metabolic transporters and more. Study of the CHI gene family and isoflavonoid biosynthesis has provided us with new insights into production and regulation of this important plant natural product. This knowledge can facilitate the manipulation of metabolic content and composition in legumes, and introduction of the isoflavonoid pathway into non-legume crops

Empagliflozin alleviates renal inflammation and oxidative stress in streptozotocin-induced diabetic rats partly by repressing HMGB1-TLR4 receptor axis

Objective(s): Empagliflozin, a sodium-glucose cotransporter-2 (SGLT-2) inhibitor, possesses verified anti-inflammatory and anti-oxidative stress effects against diabetic nephropathy. The present investigation aims to examine empagliflozin effects on the renal levels of high mobility group box-1 (HMGB1), a potent inflammatory cytokine, and its respective receptor toll-like receptor-4 (TLR-4) in STZ-induced diabetic rats.Materials and Methods: Empagliflozin at 10 mg/kg per os (p.o.) was administered for 4 weeks, starting 8 weeks after the induction of diabetes. Renal function, kidney inflammation, oxidative stress, and apoptosis markers as well as renal HMGB1, receptor for advanced glycation end products (RAGE), and TLR-4 levels were assessed.Results: In addition to down-regulating NF-κB activity in renal cortices, empagliflozin reduced renal levels of HMGB1, RAGE, and TLR-4. It alleviated renal inflammation as indicated by diminished renal expressions of inflammatory cytokines and chemokines like tumor necrosis factor-alpha (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) and also decreased urinary levels of interleukin-6 (IL-6) and alpha-1 acid glycoprotein (AGP). Moreover, empagliflozin ameliorated renal oxidative stress as demonstrated by decreased renal malondialdehyde (MDA) and elevated renal activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX). It also suppressed renal caspase-3, the marker of apoptosis; and furthermore, enhanced renal function noticed by the declined levels of serum urea and creatinine.Conclusion: These findings underline that empagliflozin is able to attenuate diabetes-related elevations in renal HMGB1 levels, an influential inflammatory cytokine released from the necrotic and activated cells, and its correspondent receptors, i.e., RAGE and TLR-4

Transcriptomic evidence for the control of soybean root isoflavonoid content by regulation of overlapping phenylpropanoid pathways

Abstract Background Isoflavonoids are a class of specialized metabolites found predominantly in legumes. They play a role in signaling for symbiosis with nitrogen-fixing bacteria and inhibiting pathogen infection. Results A transcriptomic approach using soybean cultivars with high (Conrad and AC Colombe) and low (AC Glengarry and Pagoda) root isoflavonoid content was used to find elements that underlie this variation. Two genes, encoding the flavonoid-metabolizing enzymes, flavonoid 3\u2032-hydroxylase ( GmF3\u2032H ) and dihydroflavonol 4-reductase ( GmDFR ) , had lower expression levels in high isoflavonoid cultivars. These enzymes compete with isoflavonoid biosynthetic enzymes for the important branch-point substrate naringenin and its derivatives. Differentially expressed genes, between the two sets of cultivars, encode transcription factors, transporters and enzymatic families of interest, such as oxidoreductases, hydrolases and transferases. In addition, genes annotated with stress and disease response were upregulated in high isoflavonoid cultivars. Conclusions Coordinated regulation of genes involved in flavonoid metabolism could redirect flux into the isoflavonoid branch of the phenylpropanoid pathway, by reducing competition for the flavanone substrate. These candidate genes could help identify mechanisms to overcome the endogenous bottleneck to isoflavonoid production, facilitate biosynthesis in heterologous systems, and enhance crop resistance against pathogenic infections

Retraction Note: The effects of valsartan on renal glutathione peroxidase expression in alleviation of cyclosporine nephrotoxicity in rats

This paper1 was simultaneously submitted by the authors to "Transplantation" journal under the title "The Effects of Valsartan on Renal Klotho Expression and Oxidative stress in Alleviation of Cyclosporine Nephrotoxicity" and was accepted for publication in that journal. On the basis of BioImpacts policy in accordance with the Committee on Publication Ethics (COPE), any duplicate submission is considered as an ethical misconduct. The accepted manuscript in Transplantation shared considerable overlapping text and data (identical images and materials) with the published paper1 in BioImpacts. The Editor-in-Chief of BioImpacts, Prof. Y. Omidi, was alerted by the Editor-in-Chief of Transplantation, Prof. Jeremy R. Chapman, on such duplication. A comprehensive investigation through comparison of both papers was conducted by the editorial office of BioImpacts along with the Ethics Committee of Tabriz University of Medical Sciences (TUOMS) under Prof. M. R. Rashidi, who also acts as Director-in-Charge of BioImpacts and TUOMS Vice Chancellor of Research and Technology Affairs. As a result, they decided to retract this paper in line with the COPE recommendations. The authors were informed and advised on this serious ethical breach. Therefore, the paper is retracted at the request of authors, the aforementioned committee and the Editor-in-Chief of BioImpacts, even though the corresponding author believed that two papers were different in their aims, studied animals, and factors. By the way, they apologize for any inconvenience this may have caused. One of the conditions of submission of a paper to BioImpacts is that authors declare explicitly that "This manuscript has been exclusively submitted to this journal and is not under review or accepted for publication elsewhere". As such, this paper represents abuse of the scientific publishing system. As a peer-review multidisciplinary international "Publish Free" and "Access Free" journal, BioImpacts strongly adheres to the "Publication Ethics", and its foremost goal is to preserve the integrity of the scientific reports in the highest standards, therefore the journal takes all issues of publication misconduct seriously

The effects of valsartan on renal glutathione peroxidase expression in alleviation of cyclosporine nephrotoxicity in rats

Introduction: Nephrotoxicity as a side effect caused by the immunosuppressive drug, cyclosporine-A (CsA), can be a major problem in transplant medicine. Oxidative stress may play an important role in the CsA-induced nephrotoxicity. It has been shown that the antihypertensive drug, valsartan (Val), has also renoprotective effects but, its molecular mechanism is largely unknown. In the present study, it was aimed to evaluate the Val effect in the alleviation of CsA nephrotoxicity via probable renal glutathione peroxidase (GPx) upregulation and oxidative stress decrease. Methods: Thirty-two Sprague-Dawley rats were divided into four groups based on CsA and/or Val administration: group A (Control, 1 mL/kg/day of olive oil as vehicle), group B (CsA, 30 mg/kg/day), group C (CsA+Val, 30+30 mg/kg/day), and group D (Val, 30 mg/kg/day). After the administration period (six weeks), renal GPx expression was evaluated by real-time polymerase chain reaction (PCR). Plasma levels of GPx and 8-Hydroxydeoxyguanosine (8-OHdG) were measured by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA) and protein carbonyl groups (PCG) were measured by spectrophotometer. Plasma levels of urea and creatinine were measured by an autoanalyzer. Results: CsA treatment led to the decrease in renal expression and plasma levels of GPx in comparison to other study groups. Rats received CsA were detected to have significantly (p<0.05) higher plasma 8-OHdG, MDA, PCG, urea, and creatinine levels in comparison to other groups. Plasma urea and creatinine levels were negatively correlated with renal GPx expression and positively correlated with the oxidative stress markers. Conclusion: Administration of Val may result in attenuating the nephrotoxic side effect of CsA via probable renal GPx upregulation, and subsequently oxidative stress decrease

Childhood Sex-Typed Behavior and Gender Change in Individuals with 46,XY and 46,XX Disorders of Sex Development: An Iranian Multicenter Study

Disorders of sex development (DSD) are congenital conditions in which the typical genetic and hormonal profiles are affected and thereby the usual process of sexual differentiation. Most of these studies, however, have been conducted in Western countries. In the present study, preschool sex-typed activities of Iranian individuals with DSD and their age-matched non-affected male and female relatives were assessed using the Pre-School Activities Inventory (PSAI) modified for retrospective self-report. A total of 192 individuals participated in our study, including 33 46,XX individuals with congenital adrenal hyperplasia (CAH; M age = 10.36, SD = 5.52), 15 46,XY individuals with complete androgen insensitivity syndrome (CAIS; M age = 19.8, SD = 7.14), and 16 46,XY individuals with 5-alpha reductase deficiency type-2 (5α-RD-2; M age = 17.31, SD = 7.28), as well as one age-matched non-affected male and female relative for each patient. With regard to PSAI scores, male-identifying participants with 5α-RD-2 and male controls reported similar levels of male-typical childhood play. Female-identifying participants with 5α-RD-2 and CAH showed comparable scores: significantly less masculine and more feminine than male controls, but significantly more masculine and less feminine than females with CAIS and female controls. These findings support the role of androgens in the development of sex-typical childhood play behavior, with those being exposed to higher levels of fetal functional androgens expressing more masculine behavior at preschool ages