Coordinated modification in expression levels of HSPA1A/B, DGKH, and NOTCH2 in Parkinson's patients' blood and substantia nigra as a diagnostic sign: the transcriptomes' relationship.

BACKGROUND: Diagnosis of Parkinson's disease (PD) is associated with a vast number of challenges. This study aimed to assess the overlap of PD patients' transcriptomes in the substantia nigra (SN) with peripheral blood mononuclear cells (PBMCs) to discover potential biomarkers for diagnosis. METHODS: GEO data were used to select genes with significant changes in expression level in the SN region and eligible studies. Also, transcriptome data related to blood of PD patients with other neurodegenerative diseases (ND) was considered. Differential expression genes between PD and control were evaluated in the SN and blood, and RT-qPCR was applied to validate the findings. RESULTS: At the expression level, no significant similarity in long non-coding RNA was found between the patients' SN and blood. While in silico results revealed 16 common mRNAs in SN and blood with significant expression levels. Among all overexpressed mRNAs, HSPA1A/B expression level had the highest expression difference between control and PD samples. Moreover, DGKH had the highest score of down-regulated genes in both blood and SN. The NOTCH pathway had the highest score pathway among up-regulated pathways, and the expression levels of NOTCH2, H4C8, and H2BC21 associated with this pathway had the most ability to separate the control and PD populations. Furthermore, RT-qPCR results revealed that HSPA1A/B, NOTCH2, and H4C8 were overexpressed in PD PBMCs, while DGKH expression levels were lower compared to controls. CONCLUSION: Our findings indicate that expression levels of HSPA1A/B, DGKH, and NOTCH2 could be applied as candidate biomarkers to diagnose PD patients in the SN region and PBMCs.

Evaluating the effect of basic fibroblast growth factor on the progression of NASH disease by inhibiting ceramide synthesis and ER stress-related pathways.

Non-alcoholic steatohepatitis (NASH) is associated with intrahepatic lipid accumulation, inflammation, and hepatocyte death. Several studies have indicated that high-fat diets increase ceramide synthases-6 (CerS-6) expression and a concomitant elevation of C16-ceramides, which can modulate endoplasmic reticulum (ER) stress and further contribute to the progression of NASH. Ceramide levels have reportedly been impacted by basic fibroblast growth factor (bFGF) in various diseases. This study looked into the role of bFGF on CerS6/C16-ceramide and ER stress-related pathways in a mouse model of NASH. Male C57BL/6J mice were fed a western diet (WD) combined with carbon tetrachloride (CCl4) for eight weeks. Next, bFGF was injected into the NASH mice for seven days of continuous treatment. The effects of bFGF on NASH endpoints (including steatosis, inflammation, ballooning, and fibrosis), ceramide levels and ER-stress-induced inflammation, reactive oxygen species (ROS) production, and apoptosis were evaluated. Treatment with bFGF significantly reduced CerS-6/C16-ceramide. Further, the inflammatory condition was alleviated with reduction of nuclear factor-kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6) gene expression. ROS level was also reduced. ER stress-related cell death diminished by reducing C/EBP homologous protein (CHOP) mRNA expression and caspase 3 activity. Furthermore, activation of the hepatic stellate cells was inhibited in the bFGF-treated mice by lowering the amount of alpha-smooth muscle actin (α-SMA) at the mRNA and protein level. According to our findings, CerS-6/C16-ceramide alteration impacts ER stress-mediated inflammation, oxidative stress, and apoptosis. The bFGF treatment effectively attenuated the development of NASH by downregulating CerS-6/C16-ceramide and subsequent ER stress-related pathways.

Genome-Wide Expression Analysis of Root Tips in Contrasting Rice Genotypes Revealed Novel Candidate Genes for Water Stress Adaptation.

Root system architecture (RSA) is an important agronomic trait with vital roles in plant productivity under water stress conditions. A deep and branched root system may help plants to avoid water stress by enabling them to acquire more water and nutrient resources. Nevertheless, our knowledge of the genetics and molecular control mechanisms of RSA is still relatively limited. In this study, we analyzed the transcriptome response of root tips to water stress in two well-known genotypes of rice: IR64, a high-yielding lowland genotype, which represents a drought-susceptible and shallow-rooting genotype; and Azucena, a traditional, upland, drought-tolerant and deep-rooting genotype. We collected samples from three zones (Z) of root tip: two consecutive 5 mm sections (Z1 and Z2) and the following next 10 mm section (Z3), which mainly includes meristematic and maturation regions. Our results showed that Z1 of Azucena was enriched for genes involved in cell cycle and division and root growth and development whereas in IR64 root, responses to oxidative stress were strongly enriched. While the expansion of the lateral root system was used as a strategy by both genotypes when facing water shortage, it was more pronounced in Azucena. Our results also suggested that by enhancing meristematic cell wall thickening for insulation purposes as a means of confronting stress, the sensitive IR64 genotype may have reduced its capacity for root elongation to extract water from deeper layers of the soil. Furthermore, several members of gene families such as NAC, AP2/ERF, AUX/IAA, EXPANSIN, WRKY, and MYB emerged as main players in RSA and drought adaptation. We also found that HSP and HSF gene families participated in oxidative stress inhibition in IR64 root tip. Meta-quantitative trait loci (QTL) analysis revealed that 288 differentially expressed genes were colocalized with RSA QTLs previously reported under drought and normal conditions. This finding warrants further research into their possible roles in drought adaptation. Overall, our analyses presented several major molecular differences between Azucena and IR64, which may partly explain their differential root growth responses to water stress. It appears that Azucena avoided water stress through enhancing growth and root exploration to access water, whereas IR64 might mainly rely on cell insulation to maintain water and antioxidant system to withstand stress. We identified a large number of novel RSA and drought associated candidate genes, which should encourage further exploration of their potential to enhance drought adaptation in rice.

Cytotoxic effect of hydroalcoholic extract of Cota tinctoria (L) J. Gay on AGS and Hep-G2 cancer cell lines / Efecto citotóxico del extracto hidroalcohólico de Cota tinctoria (L) J. Gay sobre líneas celulares de cáncer AGS y Hep-G2

Cota tinctoria is a medicinal plant which has been used for management of cancer in folk medicine of various regions. The aim of present study is to investigate cytotoxic activity of different concentrations of hydroalcoholic extract of C. tinctoria flowers on gastric (AGS) and liver (Hep-G2) cancer cell lines as well as Human Natural GUM fibroblast (HUGU) cells. Cell mortality rates were examined after 24, 48 and 72 h incubations using the MTT assay. IC50of extract on AGS cells after 24, 48 and 72h was 1.46, 1.29 and 1.14 µg/mL respectively. The extract demonstrated IC50 of 5.15, 3.92 and 2.89 µg/mL on Hep-G2 cells after 24, 48 and 72 h respectively. No cytotoxic effect was detected on HUGU (Human Natural GUM fibroblast) cells. C. tinctoria seems to have a promising potential to be considered as a source for anticancer drug discovery. However, more experimental and clinical studies are required.

Cota tinctoria es una planta medicinal que se ha utilizado para el tratamiento del cáncer en la medicina popular de varias regiones. El objetivo del presente estudio es investigar la actividad citotóxica de diferentes concentraciones de extracto hidroalcohólico de flores de C. tinctoria en líneas celulares de cáncer gástrico (AGS) e hígado (Hep-G2), así como en células de fibroblasto GUM humano natural (HUGU). Se examinaron las tasas de mortalidad celular después de incubaciones de 24, 48 y 72 h utilizando el ensayo MTT. La CI50 del extracto en células AGS después de 24, 48 y 72 h fue de 1,46; 1,29 y 1,14 µg respectivamente. El extracto demostró una CI50 de 5,15, 3,92 y 2,89 µg/mL en células Hep-G2 después de 24, 48 y 72 h, respectivamente. No se detectó ningún efecto citotóxico en las células HUGU (fibroblasto GUM humano natural). C. tinctoria parece tener un potencial prometedor para ser considerada como una fuente de descubrimiento de fármacos contra el cáncer. Sin embargo, se requieren más estudios experimentales y clínicos.

A fast and accurate mouse model for inducing non-alcoholic steatohepatitis.

Aim: This study aimed to perform a head-to-head comparison of changes during NASH progression throughout 6-11 weeks of an experiment to supply a faster nutritional model in mimicking NASH to decrease the duration and cost of in vivo studies. Background: New therapies are urgently needed because of the growing prevalence of non-alcoholic steatohepatitis (NASH) and the lack of an effective treatment approach. Currently, dietary interventions are the most efficient options. Methods: This study compared features of NASH in a murine model using protocol that combined special nutritional regimes based on the combination of 21.1% fat, 41% sucrose, and 1.25% cholesterol with weekly intraperitoneal injections of carbon tetrachloride (CCl4). Male C57BL/6J mice received either special compositions + CCl4 (NASH group) or standard chow diet (healthy control group) for 11 weeks. Liver histopathology based on hematoxylin and eosin (H&E) and Masson's Trichrome (TC) staining and biochemical analyses were used to assess disease progression. Results: In C57BL/6J mice administered a high fat, high cholesterol, high sucrose diet and CCl4 for 8 weeks, steatohepatitis with pronounced hepatocyte ballooning, inflammation, steatosis, and fibrosis was observed. According to the NAFLD activity scoring system, the maximum NAS score was manifested after 8-9 weeks (NAS score: 6.75). Following this protocol also led to a significant increase in AST and ALT, total cholesterol, and total triglyceride serum levels in the NASH group. Conclusion: Following the special nutritional regime based on high fat, cholesterol, and sucrose in combination with CCL4 injections resulted in a NASH model using C57BL/6J mice in a shorter time compared to similar studies. The obtained histopathological NASH features can be advantageous for preclinical drug testing.

Design and Production An Effective Bispecific Tandem Chimeric Antigen Receptor on T Cells against CD123 and Folate Receptor ß towards B-Acute Myeloid Leukaemia Blasts.

OBJECTIVE: The clinical studies of acute myeloid leukaemia (AML) revealed that antigen escaping variants cause cancer recurrence even after treatment with chimeric antigen receptor (CAR)-T cells that target a single tumour antigen. Due to the heterogeneous expression of antigens on leukaemia blasts, we hypothesized that a novel bispecific CAR, directed to the folate receptor beta (FRß)-binding single-chain variable fragment (scFv) and an IL3α-binding receptor (CD123) that has more expression in AML blasts, can decrease CAR-T cell exhaustion and increase the efficacy of CAR-T cells to prevent antigen escaping and consequent recurrence of AML. MATERIALS AND METHODS: In this experimental study, the survival, proliferation, and cytolysis of CAR-T cells remains suboptimal even with a costimulatory endodomain. Hence, we designed and constructed a tandem CAR that joins an FRß and CD123 in the second generation retroviral vector to generate a bispecific tandem CAR (TanCAR-T cell). RESULTS: TanCAR FRß-CD123 T cells showed distinct binding to FRß or CD123 expressing cells. They could lyse the leukaemia cell lines (66.1 ± 11%) comparable to the single CAR-T cells against these determinants. TanCAR FRß- CD123 T cells simultaneously engaged FRß and CD123, which promoted T cell activation in targeting and lysis of the examined leukaemia cell lines. TanCAR-T cell significantly induced interferon gamma (IFNγ) and interleukin 2 (IL-2) production more than single CAR-T cells, which produced a synergistic enhancement of TanCAR FRß-CD123 T cell function when dual antigens faced simultaneously. CONCLUSION: Dual-specific TanCAR FRß-CD123 T cells showed therapeutic potential to improve AML control by coengaging FRß and CD123 molecules in a robust, divalent immune system. This strategy may be a useful therapeutic approach in patients with relapsed B-cell malignancies.

Distal Renal Tubular Acidosis in an Iranian Patient with Hereditary Spherocytosis

Background: Hereditary spherocytosis (HS) and hereditary hereditary distal renal tubular acidosis (dRTA) are associated with mutations in the SLC4A1 gene encoding the anion exchanger 1. In this study, some patients with clinical evidence of congenital HS and renal symptoms were investigated. Methods: Twelve patients with congenital HS and renal symptoms were recruited from Ali-Asghar Children's Hospital (Tehran, Iran). A patient suspected of having dRTA was examined using whole exome sequencing method, followed by Sanger sequencing. Results: One patient (HS03) showed severe failure to thrive, short stature, frequent urinary infection, and weakness. A homozygote (rs571376371 for c.2494C>T; p.Arg832Cys) and a heterozygote (rs377051298 for c.466C>T; p.Arg156Trp) missense variant were identified in the SLC4A1 and SPTA1 genes, respectively. The compound heterozygous mutations manifested as idRTA and severe HS in patient HS03. Conclusion: Our observations, for the first time, revealed clinical and genetic characteristics of idRTA and severe HS in an Iranian patient HS03.

Integrated pan-cancer of AURKA expression and drug sensitivity analysis reveals increased expression of AURKA is responsible for drug resistance.

INTRODUCTION: The AURKA gene encodes a protein kinase involved in cell cycle regulation and plays an oncogenic role in many cancers. The main objective of this study is to analyze AURKA expression in 13 common cancers and its role in prognostic and drug resistance. METHOD: Using the cancer genome atlas (TCGA) as well as CCLE and GDSC data, the level of AURKA gene expression and its role in prognosis and its association with drug resistance were evaluated, respectively. In addition, the expression level of AURKA was assessed in colorectal cancer (CRC) and gastric cancer (GC) samples. Besides, using Gene Expression Omnibus (GEO) data, drugs that could affect the expression level of this gene were also identified. RESULTS: The results indicated that the expression level of AURKA gene in 13 common cancers increased significantly compared to normal samples or it survived poorly (HR >1, p < 0.01) in lung, prostate, kidney, bladder, and uterine cancers. Also, the gene expression data showed increased expression in CRC and GC samples compared to normal ones. The level of AURKA was significantly associated with the resistance to SB 505124, NU-7441, and irinotecan drugs (p < 0.01). Eventually, GEO data showed that JQ1, actinomycin D1, and camptothecin could reduce the expression of AURKA gene in different cancer cell lines (logFC < 1, p < 0.01). CONCLUSION: Increased expression of AURKA is observed in prevalent cancers and associated with poor prognostic and the development of drug resistance. In addition, some chemotherapy drugs can reduce the expression of this gene.

Full-length Sequencing and Genotyping of Hepatitis A Virus among Acute Hepatic Patients in Iran.

BACKGROUND: Given the lack of access to a full-length sequence of hepatitis A virus (HAV) and scarce information about its circulating genotype, sub-type and strain in Iran, two specimens were isolated from two patients with clinical symptoms of acute HAV to determine the full-length sequence of HAV. Following the phylogenetic and molecular study, we determined HAV genotype, sub-genotype, and strain of circulating virus in Iran. METHODS: According to real-time PCR results, 16 pairs of overlapped specific primers were used to determine the full-length sequence of HAV by whole-genome amplification (WGA) and using the Sanger method. Moreover, the results were assessed using Chromas, CLC Genomics Workbench, Mega 6, and RDP software. RESULTS: The full-length genome of HAV was amplified and sequenced with a length of 7,182 nucleotides. According to the obtained sequences, the phylogenetic tree of the mentioned viruses was drawn using MEGA 6 software and 44 full-genome viruses registered in the GenBank worldwide. Afterwards, the same process was repeated based on the protein sequence of VP1-P2A fragment in Iranian samples along with the other 22 registered protein sequences of GenBank to confirm the results of the full-genome phylogenetic tree. CONCLUSIONS: In this study, complete sequencing of two HAV specimens was carried out using the overlapping amplification and Sanger methods. According to the results of the phylogenetic tree, the circulating HAV in Iran had Genotype I and sub-genotype B and strain HM-175. In the present study, the full sequences of HAV of the two specimens were registered with accession numbers of BankIt 2277890/MN746031 and BankIt 2287607/MN746032.

Prodigiosin induced the caspase-dependent apoptosis in human chronic myelogenous leukemia K562 cell.

BACKGROUND AND PURPOSE: Chronic myeloid leukemia (CML) as a myeloproliferative disease is characterized by increased cellularity of bone marrow. Implementing the latest treatment protocols is currently accompanied by serious and life-threatening side effects. There are worldwide attempts to find new effective and potent therapeutic agents with minimal side effects on CML patients. This in vitro study was carried out to discover the potential antiproliferative and apoptotic effects of naturally produced prodigiosin (PDG) on K562 cells as an accepted model of CML. EXPERIMENTAL APPROACH: The anti-proliferative effect of PDG was measured by MTT assay. To highlight the mechanism of cytotoxicity, the apoptotic cell death pathway was investigated by morphological and biochemical assessments. The dual acridine orange/ethidium bromide staining technique and western blotting method were applied to assess the mechanism of the potential apoptotic impact of PDG on K562 cells. FINDINGS/RESULTS: PDG-induced time- and concentration-dependent anti-proliferative effects were revealed with an estimated IC50 value of 54.06 µM. The highest cell viability reduction (60%) was recorded in cells, which were exposed to 100 µM concentration. Further assays demonstrated that in the dual acridine orange/ethidium bromide staining method the cell population in the late apoptosis phase was increased in a concentration-dependent manner, which was confirmed with remarkable DNA fragmentation. CONCLUSION AND IMPLICATIONS: We found that the PDG-induced apoptosis in K562 cells is mediated through the caspase-3 activation both in mRNA and protein levels. Our results suggest that PDG could be a potent compound for further pharmacokinetic and pharmacodynamics studies in the in vivo model of CML.

Dysregulation of Myt1 expression acts as a potential peripheral biomarker for major depressive disorder and bipolar disorder.

Major depressive disorder (MDD) and bipolar disorder (BPD) are among the most debilitating mental conditions. Diagnostic criteria for MDD include psychological and physical symptoms, such as low mood and changes in appetite or sleep, respectively. BPD in addition to periods of depression represents episodes of mania or hypomania, and elevation in mood and energy levels are associated with this condition. Dysregulation in adult neurogenesis and myelination have been reported in psychiatric disorders. As a key factor in neurogenesis, it was hypothesized that Myt1 gene expression may be altered in these conditions. Using Real-time PCR, Myt1 expression level in 100 MDD patients and 100 BPD patients, compared with healthy control (HC) individuals was evaluated. Results demonstrate significant downregulation of Myt1 in MDD and BPD. Logistic regression analysis and binary classification evaluation reveal potential risk factor and biomarker characteristics of Myt1, respectively. Moreover, forward and backward digit span results denote a significant reduction in the function of working memory (WM) of MDD and BPD subjects. Correlation analysis revealed a significant association between Myt1 downregulation and WM disruption in the affected individuals. In conclusion, due to its altered role in neurogenesis, downregulation of Myt1 can be associated with the pathology of MDD and BPD.

Investigating the Relationship Between the Expression Level of Mucin Gene Cluster (MUC2, MUC5A, and MUC5B) and Clinicopathological Characterization of Colorectal Cancer.

Background: Colorectal cancer (CRC) is one of the most common cancers in the world and has a high mortality rate. It is accepted that dysfunction in the expression of mucins are associated with the occurrence and development of CRC. Therefore, the present study aimed to investigate the expression of MUC2, MUC5A, and MUC5B genes in CRC and their relationship with clinicopathological variables. Materials and Methods: The population included 28 patients after a colonoscopy and confirmation of the results. Tumors and parallel adjacent normal tissues from CRC patients were collected. RNA extraction and cDNA synthesis were performed using the corresponding kits. The gene primer was designed and RT-PCR was used to evaluate gene expression. The t-test and ANOVA were used to examine the differences between the different groups. Data analysis was performed using Prism8 software. Tumors from CRC patients were retrospectively collected from Taleghani Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Results: The results showed that the expression of MUC2, MUC5A, and MUC5B genes was lower in patients with CRC aged 50 years or younger than was in older patients (P<0.05). Only the MUC5B gene expression was associated with tumor grades, which was higher in poorly differentiated tumors. The expression of MUC5A and MUC2 genes was higher in stage IV of the tumor than in other stages (P<0.05). Conclusion: Among the changes in the expression of MUC secretory genes, including MUC2, MUC5A, and MUC5B and clinicopathological variables, there was a relationship that could have prognostic and diagnostic value in CRC. Conclusion: None.

The Association Between Fennel Extract, Serum Lipid Profile, and Leptin Receptor Expression.

Introduction: Obesity is among the most severe challenges of our era, with significant health consequences and a high economic burden for health systems. Therefore, many countries have developed political agendas to cope with this ever-rising challenge. Along with chemical medications developed to manage obesity, researchers have focused on some natural ingredients and herbal extracts that are effective in reducing weight. The current study investigated the association between Foeniculum vulgar (fennel) extracts and body weight, lipid profile, and leptin. Methods: In total, 35 adult male BALB/c mice were investigated in sham, fennel 50 mg/kg, fennel 100 mg/kg, and fennel 200 mg/kg (n=7) groups. The mice were administered fennel extracts for fourteen days while weighted at the intervention's beginning and end. Then, their weight, lipid profile, serum leptin, and expression of leptin protein in the hypothalamus were measured. Results: After providing the intervention, leptin receptor protein expression was increased in all groups, while serum leptin didn't change significantly. Moreover, a significant decrease was observed in the cholesterol dose of 100 mg/kg/day, triglycerides in 100 and 200 mg/kg/day, and LDL in 50 and 100 mg/kg/day. Serum HDL was increased significantly in a dose of 100 mg/kg/day. Conclusion: Fennel extract can decrease the lipid profile by changing the expression of the leptin receptor. Highlights: Obesity contributes to many health problems and dyslipidemia.Leptin is known for its hunger-blocking can regulate food intake and affects the levels of lipid. circulation.Fennel is a plant with strong antioxidant activities [18] that can influence (increase) satiety and (reduce) food intake.Fennel contains phytosterols that are known to reduce cholesterol solubilization that in turn decreases its absorption.Fennel extract can improve the lipid profile by influencing the leptin receptor expression. Plain Language Summary: Obesity is one of the most serious challenges of our era, with significant health consequences. Researchers have focused on some natural ingredients and herbal extracts. The current study aimed to investigate the association between Foeniculum vulgar (fennel) extracts and body weight, lipid profile, and leptin. After treatment of fennel, leptin receptor protein expression was increased in all groups, while serum leptin didn't change significantly. A significant decrease was observed in the cholesterol, triglycerides and LDL in some doses of fennel. Serum HDL was increased significantly in a dose of 100 mg/kg/day. So fennel extract can decrease the lipid profile by changing the expression of the leptin receptor.

Selenium nanoparticles induced variations in growth, morphology, anatomy, biochemistry, gene expression, and epigenetic DNA methylation in Capsicum annuum; an in vitro study.

This study aimed to explore whether supplementation of the culture medium with selenium nanoparticles (nSe) can influence growth, biochemistry, expression of transcription factors, and epigenetic DNA methylation in Capsicum annuum. The seeds were grown in hormone-free MS culture medium supplemented with nSe (0, 0.5, 1, 10, and 30 mgL-1) or corresponding doses of bulk type selenate (BSe). Incorporation of nSe into the medium caused variations in morphology and growth in a manner dependent on the dose and Se type. The low doses of nSe displayed growth-promoting effects, whereas nSe at 10 and 30 mgL-1 were associated with severe toxicity and abnormality in leaf and root development. MSAP analysis confirmed the substantial variation in cytosine DNA methylation in response to the toxic dose of nSe exhibiting epigenetic modification. The nSe toxicity was associated with DNA hyper-methylations. The nSe treatments transcriptionally upregulated the bZIP1 transcription factor by an average of 3.5 folds. With a similar trend, the upregulation (mean = 9.8 folds) in the expression of the WRKY1 transcription factor resulted from the nSe application. The nSe0.5 or nSe1 treatments resulted in a significant induction (mean = 48%) in nitrate reductase activity. A high dose of nSe led to an increase in proline concentration. The nSe treatments were also associated with modifications in activities of peroxidase and catalase enzymes. Besides, the nSe utilization increased the activity of phenylalanine ammonia-lyase enzyme (mean = 76%) and concentrations of soluble phenols (mean = 51%). The toxic dose of nSe also caused abnormalities in the structure of the stem apical meristem. The nSe toxicity was also associated with inhibition in the differentiation of xylem tissues. These findings provide novel insights into the nSe-associated molecular variations in conferring the modified growth, anatomy, and metabolism.

TWIST1 upregulates matrix metalloproteinase (MMP) genes family in esophageal squamous carcinoma cells.

Twist-related protein 1 (TWIST1), a highly conserved basic helix-loop-helix transcription factor, stimulates epithelial-mesenchymal transition (EMT) and plays a crucial role in the regulation of the extracellular matrix (ECM) and cell-cell adhesion. Our aim in this study was to evaluate the functional correlation between TWIST1 and MMP genes in human ESCC cell lines, KYSE-30 and YM-1. To generate recombinant retroviral particles, the Pruf-IRES-GFP-hTWIST1 was co-transfected into HEK293T along with pGP and pMD2. G as well as Pruf-IRES-GFP control plasmid. Stably transduced high-expressing GFP-hTWIST1 and GFP-control KYSE-30 cells were generated. The produced retroviral particles were transduced into the KYSE-30 and YM-1 ESCC cells. Ectopic expression of TWIST1 mRNA and expression of the MMP genes (MMP-2, MMP-3, MMP-7, MMP-9, and MMP-10) were examined by relative comparative real-time PCR. In silico analysis of the MMP markers and their promoter elements was explored. Moreover, the scratch wound assay was used to evaluate the migration of TWIST1-induced cells. TWIST1 level was up-regulated by nearly 5-fold and 7.4-fold in GFP-hTWIST1 KYSE-30 and YM-1 cells compared to GFP control cells, respectively. Interestingly, this enforced expression of TWIST1 subsequently caused significant overexpression of transcripts for selected MMP genes in GFP-hTWIST1 in comparison with GFP control cells in both ESCC cell lines. Also, the scratch assay indicated that TWIST1 expression effectively increased the migration of GFP-TWIST1 KYSE-30 cells against GFP KYSE-30 control cells in vitro. The present findings illuminate that TWIST1 may contribute broadly to ESCC development in concert with up-regulation of MMPs expression and further suggest the potential advantage of exerting TWIST1/MMPs signaling axis as a framework from which to expand our understanding about the mechanisms of ESCC tumorigenesis.

Red elemental selenium nanoparticles mediated substantial variations in growth, tissue differentiation, metabolism, gene transcription, epigenetic cytosine DNA methylation, and callogenesis in bittermelon (Momordica charantia); an in vitro experiment.

To gain a better insight into the selenium nanoparticle (nSe) benefits/toxicity, this experiment was carried out to address the behavior of bitter melon seedlings to nSe (0, 1, 4, 10, 30, and 50 mgL-1) or bulk form (selenate). Low doses of nSe increased biomass accumulation, while concentrations of 10 mgL-1 and above were associated with stem bending, impaired root meristem, and severe toxicity. Responses to nSe were distinct from that of bulk in that the nano-type exhibited a higher efficiency to stimulate growth and organogenesis than the bulk. The bulk form displayed higher phytotoxicity than the nano-type counterpart. According to the MSAP-based analysis, nSe mediated substantial variation in DNA cytosine methylation, reflecting the epigenetic modification. By increasing the concentration of nSe, the expression of the WRKY1 transcription factor linearly up-regulated (mean = 7.9-fold). Transcriptions of phenylalanine ammonia-lyase (PAL) and 4-Coumarate: CoA-ligase (4CL) genes were also induced. The nSe treatments at low concentrations enhanced the activity of leaf nitrate reductase (mean = 52%) in contrast with the treatment at toxic concentrations. The toxic concentration of nSe increased leaf proline concentration by 80%. The nSe supplement also stimulated the activities of peroxidase (mean = 35%) and catalase (mean = 10%) enzymes. The nSe-treated seedlings exhibited higher PAL activity (mean = 39%) and soluble phenols (mean = 50%). The nSe toxicity was associated with a disrupted differentiation of xylem conducting tissue. The callus formation and performance of the explants originated from the nSe-treated seedlings had a different trend than that of the control. This experiment provides new insights into the nSe-associated advantage/ cytotoxicity and further highlights the necessity of designing convincing studies to introduce novel methods for plant cell/tissue cultures and agriculture.

Effect of treadmill exercise on catalepsy and the expression of the BDNF gene in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine -induced Parkinson in male NMRI mice.

OBJECTIVES: It is known that treadmill exercise has beneficial effects on the nervous system. The brain-derived neurotrophic factor (BDNF) plays a role in such effects. This study aimed at investigating effects of intermittent treadmill exercise-induced behavioral, histology, and immunohistochemistry (H&E, TH) measurement of brain interleukin-10 (IL-10) in a mice male model of Parkinson's disease (PD), which is induced by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), as well as the role of BDNF gene in exercise effects. MATERIALS AND METHODS: The animals were divided into Control (C), Saline (S), Parkinson (P), Exercise (E), and Parkinson + Exercise (PE) groups. Bar test was performed for the 21-day protocol with 5 days a week treadmill exercises. In this regard, brains were removed from the skull for H&E, TH, IL-10, and the expression of the BDNF gene using the MPTP male mice PD model. RESULTS: MPTP reduced the number of DA neurons in the substantia nigra (SNpc), whereas daily exercise administration on 1st, 7th, 14th, and 21st days significantly reduced the catalepsy duration induced by MPTP. The results of H&E and TH studies showed that MPTP significantly reduced the number of TH+ neurons in the SNpc compared with those of the control mice. The MPTP caused a marked decrease in basal protein levels of IL-10 in SNpc and corpus striatum in the Parkinson (P) group as compared with controls. Treatment with Exercise (E) group had the most BDNF expression (3.71), and the Parkinson (P) group also had the least BDNF expression (0.18) relative to controls. CONCLUSION: The treadmill exercise having neuroprotective effects in SNpc and corpus striatum has improved MPTP associated with motor deficits. It is considered as a non-pharmacological tool for the management of PD.

The effects of UV-B radiation on genetic and biochemical changes of Pelargonium graveolens L'Her.

Ultraviolet radiation induces biochemical and genetic changes in plants. The aim of this study was to investigate the effects of UV-B radiation on genetic stability, phenolic compounds and antioxidant activity of Pelargonium graveolens L'Her. Plant cuttings were exposed to 0, 0.12. 0.26 and 0.38 W/m2 of UV-B radiation. Results indicated that by increasing the UV-B radiation intensity, total phenols, flavonoids and anthocyanin contents, Phenylalanine ammonia lyase activity and antioxidant capacity were increased. Analysis of four flavonols (quercetin, myricetin, kaempferol and rutin) contents of leaves extract by HPLC indicated that these four flavonols were enhanced in all treated plants and also the ratio of quercetin to kaempferol (Q/K) showed a significant increase (P ≤ 0.05) in UV-B treated plants in compare to control. To evaluate the genetic variation in treated plants, 10 ISSR primers were used. The highest level of percentage of polymorphism (P%), Shannon index (I), number of effective allele (Ne) and Nei' genetic diversity (He), were observed at the highest UV-B radiation (0.38 W/m2). The AMOVA analysis also showed a significant genetic differentiation (P ≥ 0.001) among the studied groups, and confirmed the differentiation of groups obtained by the cluster analysis of molecular data. Overall, these results showed that biochemical changes in different intensities of UV-B were in line with genetic variations, so that the highest biochemical and genetic variations were observed in 0.38 W/m2 treatment.

Differential adaptation strategies to different levels of soil water deficit in two upland and lowland genotypes of rice: a physiological and metabolic approach.

BACKGROUND: Upland genotypes of rice are less sensitive to soil water deficit (SWD), making them suitable candidates for revealing the strategies underlying plant tolerance. The physiological factors, the biochemical traits needed to withstand oxidative stress, and the metabolite fluctuations of an upland genotype (Azucena) and an intolerant lowland genotype (IR64) genotype were measured under two levels of SWD (withholding water for 7- or 14 days) to identify SWD-responsive strategies associated with tolerance. RESULTS: After withholding water for 7 days, no significant changes in physiological and biochemical traits of Azucena were observed, whereas in IR64, significant decreases in physiological factors were recorded along with increases in oxidative-stress indicators. However, the root length of Azucena increased significantly, showing a clear stress avoidance strategy. Under a prolonged treatment (14 days), IR64 entered an oxidative-damage stage, whereas Azucena exhibited a highly efficient antioxidant system. Our metabolite analysis also revealed two different enriched pathways. After a 7-day SWD, the sugar levels were decreased in the leaves of Azucena but increased in IR64. The reduction in the sugar levels (up to 1.79-log2FC) in the Azucena leaves may be indicative of their transport to the roots, supplying the carbon source needed for root elongation. Under a 14-day treatment, proline and aspartate family members accumulated to the highest levels in Azucena, whereas an increase in the levels of aromatic amino acids with key roles in the biosynthesis of secondary metabolites was detected in IR64. CONCLUSION: The adaptation strategies identified in two types of rice genotypes in confronting SWD may assist researchers in finding the proper indicators for screening more tolerant genotypes. © 2019 Society of Chemical Industry.

The effects of the electromagnetic fields on the biochemical components, enzymatic and non-enzymatic antioxidant systems of tea Camellia sinensis L.

The electromagnetic fields (EMFs) by wide range of the frequency spectrum, have capability to cause crucial alternation and deleterious effects in biological systems. The aim of the present study is to assay the biochemical components, enzymatic and non-enzymatic antioxidant systems of the electromagnetic fields treated samples of tea which is the most ancient non-alcoholic drink, containing different types of flavonols. Rutin, Quercetin, Myricetin, and Kaempferol as flavonoid components markers are also to be analyzed using high-performance liquid chromatography. The results show that The EMF's treatments brought about distinct alternations in biochemical components of tea, so that regardless of the intensity of the EMF's, less duration of exposure (30 min) caused more content of those mentioned flavonoid components (except Myricetin) than that of 60 min of exposure. A 30 min of 4 miliTesla (mT) exposure of the EMF's, resulted in the highest amount of Rutin, Quercetin, Myricetin, and Kaempferol. It is concluded that less duration of the EMF's treatments induces more production and also accumulation of enzymatic and non-enzymatic antioxidant components. In higher intensity of the EMF's (more than 4 mT), the concentrations of the mentioned biochemical components decreased.