Introduction of miR-3613-3p as a regulator of transforming growth factor-ß (TGF-ß) signaling pathway in colorectal cancer.

INTRODUCTION: Colorectal cancer (CRC) is the second common cancer and the fourth major reason of cancer death worldwide. Dysregulation of intracellular pathways, such as TGF-ß/SMAD signaling, contributes to CRC development. MicroRNAs (miRNAs) are post-transcriptional regulators that are involved in CRC pathogenesis. Here, we aimed to investigate the effect of miR-3613-3p on the TGF-ß /SMAD signaling pathway in CRC. METHODS & RESULTS: Bioinformatics analysis suggested that miR-3613-3p is a regulator of TGF-Β signaling downstream genes. Then, miR-3613-3p overexpression was followed by downregulation of TGF-ßR1, TGF-ßR2, and SMAD2 expression levels, detected by RT-qPCR. Additionally, dual luciferase assay supported the direct interaction of miR-3613-3p with 3'UTR sequences of TGF-ßR1 and TGF-ßR2 genes. Furthermore, reduced SMAD3 protein level following the miR-3613-3p overexpression verified its suppressive effect against TGF-ß signaling in HCT-116 cells, detected by western blot analysis. Finally, miR-3613-3p overexpression induced sub-G1 arrest in HCT116 cells, detected by flow cytometry, and promoted downregulation of cyclin D1 protein expression, which was detected by western blotting analysis. CONCLUSION: Our findings indicated that miR-3613-3p plays an important role in CRC by targeting the TGF-ß/SMAD signaling pathway and could be considered as a new candidate for further therapy investigations.

OCC-1D regulates Wnt signaling pathway: potential role of long noncoding RNA in colorectal cancer.

BACKGROUND: Aberrant activation of the Wnt signaling pathway is observed in most colorectal cancers (CRC). OCC-1D is a splice variant of OCC-1 gene which is considered as a long noncoding RNA (lncRNA) due to lacking the translational initiation codon of the gene. Here, we sought supporting evidence for the effects of OCC-1D on the Wnt pathway and cell cycle progression in CRC. METHODS AND RESULTS: TOP/FOPflash assay and qRT-PCR indicated that expression alterations of OCC-1D could change Wnt signaling activity in colon cancer cells. Consistently, immunocytochemistry results showed the effect of OCC-1D overexpression on nuclear localization of ß-catenin proteins in SW480 cells. Flow cytometry, wound healing and MTT assay confirmed the cell cycle stimulatory effects of OCC-1D in CRC-originated cell lines (SW480 and HCT116). qRT-PCR revealed a positive correlation between the expression level of OCC-1D and its neighboring gene, APPL2. Two distinct tests, downregulation of APPL2 mRNA by using shRNA and Wnt signaling inhibition by using small molecule, along with OCC-1D overexpression confirmed that OCC-1D lncRNA exerts its effect on Wnt signaling pathway through expression modulation of APPL2 gene. CONCLUSIONS: Collectively, we suggested the putative regulatory effects of OCC-1D lncRNA on cell cycle progression and Wnt signaling activation through enhancing the APPL2 gene transcription.

YWHAE long non-coding RNA competes with miR-323a-3p and miR-532-5p through activating K-Ras/Erk1/2 and PI3K/Akt signaling pathways in HCT116 cells.

YWHAE gene product belongs to the 14-3-3 protein family that mediates signal transduction in plants and mammals. Protein-coding and non-coding RNA (lncRNA) transcripts have been reported for this gene in human. Here, we aimed to functionally characterize YWHAE-encoded lncRNA in colorectal cancer-originated cells. RNA-seq analysis showed that YWHAE gene is upregulated in colorectal cancer specimens. Additionally, bioinformatics analysis suggested that YWHAE lncRNA sponges miR-323a-3p and miR-532-5p that were predicted to target K-Ras 3'UTR sequence. Overexpression of YWHAE lncRNA resulted in upregulation of K-Ras gene expression, while overexpression of both miR-323a-3p and miR-532-5p had an inverse effect, detected by RT-qPCR. Consistently, western blot analysis confirmed that YWHAE lncRNA overexpression upregulated K-Ras/Erk1/2 and PI3K/Akt signaling pathways, while miR-323a-3p and miR-532-5p overexpression suppressed both pathways in HCT116 cells. Furthermore, dual luciferase assay validated the direct interaction of miR-323a-3p and miR-532-5p with K-Ras 3'UTR sequence and supported the sponging effect of YWHAE lncRNA over both miRNAs. These results suggested YWHAE lncRNA as an oncogene that exerts its effect through sponging miR-323a-3p and miR-532-5p and in turn, upregulates K-Ras/Erk1/2 and PI3K/Akt signaling pathways. Consistently, flow cytometry analysis, MTT assay and measuring cyclin D1 gene expression, confirmed the cell cycle stimulatory effect of YWHAE lncRNA, while miR-323a-3p and miR-532-5p showed an inhibitory effect on cell cycle progression. Finally, wound-healing assay supported the cell migratory effect of YWHAE lncRNA in HCT116 cells. This study identified a novel mechanism involving YWHAE-encoded lncRNA, miR-323a-3p and miR-532-5p in regulating HCT116 cell survival and suggested a potential therapeutic avenue for colorectal cancer.

Hsa-miR-186-5p regulates TGFß signaling pathway through expression suppression of SMAD6 and SMAD7 genes in colorectal cancer.

TGFß signaling is a known pathway to be involved in colorectal cancer (CRC) progression and miRNAs play crucial roles by regulating different components of this pathway. Hence, finding the link between miRNAs and the pathway could be beneficial for CRC therapy. Array data indicated that miR-186-5p is a differentially expressed miRNA in colorectal Tumor/Normal tissues and bioinformatics tools predicted SMAD6/7 (inhibitory SMADs) as bona fide targets of this miRNA. Here, we intended to investigate the regulatory effect of the miR-186-5p expression on TGFß signaling in CRC. Firstly, the miR-186-5p overexpression in HCT116 cells resulted in a significant reduction of SMAD6/7 expression, measured through RT-qPCR. Then, the direct interactions of miR-186-5p with SMAD6/7 3'UTRs were supported through dual luciferase assay. Furthermore, miR-186-5p overexpression suppressed proliferation, cell viability, and migration while, it increased apoptosis in CRC cells, assessed by cell cycle, MTT, scratch and Annexin V/PI apoptosis assays. Consistently, miR-186-5p overexpression resulted in reduced CyclinD1 protein using western blot, and also resulted in increased P21 and decreased c-Myc expression. Overall, these results introduced miR-186-5p as a cell cycle suppressor through downregulation of SMAD6/7 expression. Thus, miR-186-5p might be served as a novel tumor suppressive biomarker and therapeutic target in CRC treatment.

Physiological, biochemical, and molecular responses of Linum album to digested cell wall of Piriformospora indica.

Plants synthesize a variety of metabolites in response to biotic elicitors. To comprehend how the digested cell wall of Piriformospora indica affects the response of ROS burst, antioxidant enzymes, amino acids profiling, and phenylpropanoid compounds such as lignans, phenolic acids, and flavonoids in Linum album hairy roots; we accomplished a time-course analysis of metabolite production and enzyme activities in response to CDCW and evaluated the metabolic profiles. The results confirms that CDCW accelerates the H2O2 burst and increases SOD and GPX activity in hairy roots. The HPLC analysis of metabolic profiles shows that the H2O2 burst shifts the amino acids, especially Phe and Tyr, fluxes toward a pool of lignans, phenolic acids, and flavonoids through alterations in the behavior of the necessary enzymes of the phenylpropanoid pathway. CDCW changes PAL, CCR, CAD, and PLR gene expression and transiently induces PTOX and 6MPROX as the main-specific products of PAL and PLR genes expression. The production of phenolic acids (e.g., cinnamic, coumaric, caffeic, and salicylic acid) and flavonoids (e.g., catechin, diosmin, kaempferol, luteolin, naringenin, daidzein, and myricetin) show different behaviors in response to CDCW. In conclusion, our observations show that CDCW elicitation can generate H2O2 molecules in L. album hairy roots and consequently changes physiological, biochemical, and molecular responses such as antioxidant system and the specific active compounds such as lignans. Quantification of metabolic contents in response to CDCW suggests enzyme and non-enzyme defense mechanisms play a crucial role in L. album hairy root adaptation to CDCW. A summary revealed that the correlation between H2O2 generation and L. album hairy root defense system under CDCW. Increase of H2O2 generation led plant to response against oxidative conditions. SOD, and GPX modulated H2O2 content, Phe, and Tyr shifted to the phenylpropanoid compounds as a precursor of PAL and TAL enzyme, the predominant phenylpropanoid compounds controlled oxidative conditions, and the other amino acids responsible for amino acid synthesis and development stages.

The urgent need for integrated science to fight COVID-19 pandemic and beyond.

The COVID-19 pandemic has become the leading societal concern. The pandemic has shown that the public health concern is not only a medical problem, but also affects society as a whole; so, it has also become the leading scientific concern. We discuss in this treatise the importance of bringing the world's scientists together to find effective solutions for controlling the pandemic. By applying novel research frameworks, interdisciplinary collaboration promises to manage the pandemic's consequences and prevent recurrences of similar pandemics.

Differential expression of STAT3 gene and its regulatory long non-coding RNAs, namely lnc-DC and THRIL, in two eastern Iranian ethnicities with multiple sclerosis.

OBJECTIVE: Genome-wide association studies (GWASs) revealed that variants of STAT3 are associated with multiple sclerosis (MS) risk. There are several studies showing the effect of ethnicity and genetic background on the characteristics of MS. Here, we aimed to investigate STAT3 gene expression status along with its two regulatory long non-coding RNAs, lnc-DC and THRIL, in order to compare the expression of these target genes among two different ethnicities in the east of Iran. METHODS: A case-control study was performed between two groups of MS populations in east of Iran. We recruited individuals with Kurdish ethnicity from North Khorasan and Sistani ethnicity from southeast of Iran. The peripheral blood mononuclear cells were obtained from all participants, and total RNA was extracted. The gene expression of the selected genes was evaluated by qPCR. RESULTS: The expression of THRIL in North Khorasan MS patients was significantly higher than controls (P = 0.03). The results of simultaneous analysis of expression of the target genes (STAT3, THRIL, and lnc-DC) in both ethnic groups failed to show any significant difference between the MS patients and controls (P > 0.05). In addition, the expression of STAT3 and THRIL genes in Sistani MS patients was statistically meaningful lower than healthy controls (P < 0.05). CONCLUSION: To our knowledge, this is the first study that compared the expression of the STAT3 gene and its regulatory molecules between two ethnic groups of Iranian MS patients. We suggested that STAT3 and its associated molecules might be differentially expressed and regulated in MS patients with different genetic background.

Optical biosensing of Streptococcus agalactiae based on core/shell magnetic nanoparticle-quantum dot.

An immunomagnetic optical probe based on a core/shell magnetic nanoparticle-quantum dot was fabricated for detection of Streptococcus agalactiae, the causative agent of pneumonia and meningitis in newborns. The silica-coated magnetic nanoparticles conjugated with anti-S. agalactiae monoclonal antibody provided high specificity for pre-enrichment of bacteria from biological samples with a complex matrix such as milk. Compared with conventional methods such as culture and molecular techniques, the combination of fluorescent quantum dot and magnetic nanoparticle enhanced the sensitivity and speed of bacterial identification. The bio-functionalized fluorescent-magnetic nanoparticles were characterized by TEM, SEM, VSM, XRD, DLS, and FTIR and applied to the detection of S. agalactiae with a limit of 10 and 102 CFU/mL in PBS and milk, respectively. This immunomagnetic optical probe can be used for rapid isolation, sensitive, and specific detection of targeted bacteria without any treatment in clinical and animal samples in the presence of other infectious agents.

Effect of single nucleotide polymorphisms in SEPS1 and SEPP1 on expression in the protein level in metabolic syndrome in subjects with cardiovascular disease.

Metabolic syndrome (MetS) results from the interaction between environmental and genetic factors. Several previous studies considered the role of selenium in developing MetS. Two selenoproteins, selenoprotein S (SelS), and the Selenoprotein P (SePP) play an important role in antioxidative defense and therefore susceptibility to MetS. The involvement of SNPs in SEPP1 and SEPS1 have not been studied in MetS subjects. This study aims to investigate the association between the risk of MetS and four polymorphisms SEPS1 (rs28665122, rs4965373), SEPP1 (rs7579, rs3877899) in an Iranian population. The sample of this case-control study consisted of 132 Iranian patients with cardiovascular disease (71 MetS and 65 non-MetS subjects) from December 2015 to March 2016. Demographic data, medical history, and para-clinical were measured, and Taqman probes were used for allelic discrimination. The level of the SelS and the SePP were measured by the ELIZA method. No significant differences were found in the genotype frequencies of SEPS1 (rs4965373, rs28665122), SEPP1 (rs7579, rs3877899) in patients with MetS and the non-MetS group. The mean of SelS in MetS subjects with SEPS1 (rs4965373) GG genotype is significantly lower than the non-MetS group (4496.99 ± 3688.5 vs. 6148.6 ± 1127.0, P = 0.009). The mean of SePP in MetS subjects with SEPP1 (rs3877899) GG genotype is significantly lower than the non-MetS group (40.73 ± 8.44 vs.83.91 ± 21.33, P = 0.002). The mean of SePP in MetS subjects with SEPP1 (rs7579) GG genotype is lower than the non-MetS group (55.52 ± 16.7 vs. 109.48 ± 29.78, P = 0.01). In summary, the results of this study does not indicate significant differences in the SEPP1 (rs7579, rs3877899) and SEPS1 (rs4965373, rs28665122) genotypes between MetS and non-MetS subjects. However, the results show that the mean of expression of SelS and SePP decreased in the subjects with SEPP1 (rs7579) GG and SEPP1 (rs3877899) GG.

A sensitive gold-nanorods-based nanobiosensor for specific detection of Campylobacter jejuni and Campylobacter coli.

BACKGROUND: Campylobacteriosis is a zoonotic infectious disease that can be mostly undiagnosed or unreported due to fastidious Campylobacter species. The aim of this study was to develop a simple, sensitive, and quick assay for the detection of Campylobacter spp. and taking advantage of the great sensitivity of gold nanorods (GNRs) to trace changes in the local environment and interparticle distance. METHODS: Characterized GNRs were modified by specific ssDNA probes of cadF gene. First, the biosensor was evaluated using recombinant plasmid (pTG19-T/cadF) and synthetic single-stranded 95 bp gene, followed by a collection of the extracted DNAs of the stool samples. The sensing strategy was compared by culture, PCR, and real-time PCR. RESULTS AND DISCUSSION: Analysis of 283 specimens showed successful detection of Campylobacter spp. in 44 cases (16%), which was comparable to culture (7%), PCR (15%), and real-time PCR (18%). In comparison with real-time PCR, the sensitivity of the biosensor was reported 88%, while the specificity test for all assays was the same (100%). However, it was not able to detect Campylobacter in 6 positive clinical samples, as compared to real-time PCR. The limit of detection was calculated to be the same for the biosensor and real-time PCR (102 copy number/mL). CONCLUSIONS: Taking high speed and simplicity of this assay into consideration, the plasmonic nanobiosensor could pave the way in designing a new generation of diagnostic kits for detection of C. jejuni and C. coli species in clinical laboratories.

HOTAIR but not ANRIL long non-coding RNA contributes to the pathogenesis of multiple sclerosis.

Studies have revealed that dysregulation in gene expression is one of the main aspects of multiple sclerosis (MS) pathogenesis. Although the molecular pathways underlying the immunomodulatory role of vitamin D (VD) in MS is not completely elucidated, VD has more recently become a topic of interest in immune regulation and is widely administered to patients with MS as an immunomodulatory supplement. Long non-coding RNAs (lncRNAs) are known to play important roles in regulation of gene expression via different mechanisms. Given that VD-related genes are regulated by epigenetic mechanisms, here we aimed to evaluate the role of VD in combination with HOTAIR and ANRIL lncRNAs using in vivo, in vitro and in silico experiments in MS pathogenesis. Our data revealed that HOTAIR but not ANRIL lncRNA is probably involved in the pathogenesis of MS and experimental autoimmune encephalomyelitis through an unclear mechanism and it seems that by affecting the expression, inflammation and VD can influence HOTAIR-related mechanisms, which require further study.

Effects of selenium supplementation on expression of SEPP1 in mRNA and protein levels in subjects with and without metabolic syndrome suffering from coronary artery disease: Selenegene study a double-blind randomized controlled trial.

Selenoprotein P (SePP) is involved in the protection against diseases. The present study is the first investigation of the effect of selenium supplementation on plasma selenium and expression of SEPP1 in mRNA and protein levels based on metabolic syndrome (MetS), in individuals suffering from coronary artery diseases. In this clinical trial, 160 patients with angiographically documented stenosis of more than 75% in each vessel were enrolled. Patients received either 200-mg selenium yeast tablets or placebo tablets orally after a meal, once daily for 60 days. The mRNA and protein levels of the selenium and SePP1 products were determined before and after the study. From the initial 160 participants, 145 subjects (71 MetS-affected individuals, 74 MetS-unaffected individuals) enrolled in this study. Comparing the selenium and placebo groups, no significant percentage changes of plasma selenium, △Ct SEPP1, or SePP were shown (P > 0.05). Moreover, beyond a significant difference for the expression of SePP in the selenium group compared to its baseline level (P < 0.05), no other significant differences were revealed for plasma selenium and △Ct SEPP1 after the intervention in either group (P > 0.05). Selenium supplementation did not affect plasma selenium or the mRNA or protein level of SePP in either groups after a 2-months intervention beyond a significant increase of SePP in the MetS group. This trial suggests that further studies should investigate the long-term use of selenium supplementation and the effect of a SePP increase on MetS as a potential therapeutic effect.

Piriformospora indica cell wall modulates gene expression and metabolite profile in Linum album hairy roots.

MAIN CONCLUSION: Elicitation of Linum album hairy roots by Piriformospora indica cell wall induced the target genes and specific metabolites in phenylpropanoid pathway and shifted the amino acid metabolism toward the phenolic compound production. Plants have evolved complex mechanisms to defend themselves against various biotic stresses. One of these responses is the production of metabolites that act as defense compounds. Manipulation of plant cell cultures by biotic elicitors is a useful strategy for improving the production of valuable secondary metabolites. This study focused on hairy root culture of Linum album, an important source for lignans. The effects of cell wall elicitor extracted from Piriformospora indica on phenylpropanoid derivatives were evaluated to identify metabolic traits related to biotic stress tolerance. Significant increases in lignin, lignans; lariciresinol, podophyllotoxin, and 6-methoxy podophyllotoxin; phenolic acids: cinnamic acid, ferulic acid, and salicylic acid; flavonoids: myricetin, kaempferol, and diosmin were observed in response to the fungal elicitor. In addition, the gene expression levels of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, cinnamoyl-CoA reductase, and pinoresinol-lariciresinol reductase significantly increased after elicitation. The composition of free amino acids was altered under the elicitation. Phenylalanine and tyrosine, as precursors of phenylpropanoid metabolites, were increased, but alanine, serine, and glutamic acid significantly decreased in response to the fungal elicitor, suggesting that the amino acid pathway may be shifted toward biosynthesis of aromatic amino acids and precursors of the phenylpropanoid pathway. These results provided evidence that up-regulation of genes involved in the phenylpropanoid pathway in response to the fungal elicitor resulted in enhanced metabolic responses associated with the protection in L. album. This approach can also be applied to improve lignan production.

Overexpressed in colorectal carcinoma gene (OCC-1) upregulation and APPL2 gene downregulation in breast cancer specimens.

Breast cancer is the most common cancer type and the second cause of cancer death in women. Different mechanisms are contributed to the initiation and progression of the breast cancer. OCC-1 and APPL2 neighboring genes located in 12q.23.3 human chromosome region are related to colorectal cancer. Here, we intended to investigate OCC-1 newly reported transcript variants and APPL2 gene expression alteration in breast cancer specimens and investigate OCC-1 variants overexpression effect on APPL2 and on cell cycle status. Rt-qPCR analysis indicated that the expression level of OCC-1A/B and OCC-1D (not OCC-1C) transcript variants has been increased while, APPL2 gene expression level has been decreased in breast cancer specimen, compared to their normal pairs. Therefore, a negative correlation of expression is evident between APPL2 and OCC-1 genes in breast cancer specimen. Unlike OCC-1A/B which encodes a small protein, OCC-1D noncoding RNA overexpression lead to APPL2 downregulation in MCF7 cells. Consistently, OCC-1D overexpression resulted in increased sub-G1 cell population in MCF7 cells, detected by flow cytometry. Altogether, these results suggest that OCC1-D variant have an inhibitory effect on APPL2 expression and may regulate the cell cycle status.

Size of single-wall carbon nanotube affects the folate receptor-mediated cancer cell targeting.

Advances in nanobiotechnology and targeting strategy could improve the delivery of therapeutic molecules into cancer cells, leading to improved treatment efficiency with minimal side effects on normal cells. To design an efficient nanocarrier, consideration of parameters that facilitate direct drug delivery into the target cells is important. We studied the effect of single-wall carbon nanotubes (SWNTs) size on their cell internalization level via the folate receptor-mediated pathway through folic acid targeting. Folate-SWNTs were covalently synthesized and characterized. Folate-SWNTs ≤ 450 nm had lower cell internalization level than folate-SWNTs >450 nm with a P value of ≤0.01. This indicated that using folate-SWNT with an average length of ≤450 nm was not suitable for receptor-mediated cancer cell targeting. Receptor-mediated uptake of folate-SWNTs is dependent on the nanoparticle length. However, sub-450 nm SWNTs could serve as a vehicle to transfer nucleic acids into the cells due to direct cell penetrance based on their needle-like structure. We find that SWNTs larger than 450 nm were suitable to target the cells through receptors. These results might provide a promising approach for designing more effective targeted delivery systems based on SWNTs.

Association of R279Q and C1562T polymorphisms of matrix metalloproteinase 9 gene and increased risk for myocardial infarction in patients with premature coronary artery disease.

BACKGROUND: A number of matrix metalloproteinase (MMP) gene polymorphisms has been identified which may be probably related to premature myocardial infarction (MI). OBJECTIVE: We assessed the relationship between the two polymorphisms of the MMP9 gene including R279Q and C1562T and occurrence of premature MI. METHODS: The study has two phases including a case-control study as the first phase and cohort study as the second phase. Initially, 1000 patients with premature coronary artery disease were classified into MI and non-MI groups. Genotyping of the polymorphism was conducted by PCRRFLP and high-resolution melting techniques. Given the two conditions of patients residing in Tehran and faced with their first episode of MI, 640 of 1000 study samples previously followed up with a median follow-up time of 45.74 months were assessed in a retrospective cohort phase regarding long-term major adverse cardiac events (MACE). RESULTS: The prevalence of wild, heterozygous, and mutant genotypes of R279Q polymorphism in MI group was 14.5%, 57.3%, and 28.2% and in non-MI group was 36.9%, 38.4%, and 24.7%, respectively, with a considerable difference (P<.001). There was a significant difference in the prevalence of wild, heterozygous, and mutant genotypes of C1562T polymorphisms in MI group (12.4%, 41.2%, and 46.4%, respectively) and in non-MI group (46.8%, 38.6%, and 14.7%, respectively; P<.001). No difference was found in total MACE-free survival rate between genotypes of R279Q and C1562T polymorphisms. CONCLUSION: C1562T and R279Q polymorphisms are associated with the susceptibility to premature MI, but cannot predict long-term cardiac events in these patients.

The expression of pluripotency and neuronal differentiation markers under the influence of electromagnetic field and nitric oxide.

Nitric oxide (NO) is a diatomic free radical compound that as a secondary messenger contributes to cell physiological functions and its variations influence proteins activity and triggering intracellular signaling cascades. Low frequency electromagnetic field (EMF) alters the cell biology such as cell differentiation by targeting the plasma membrane and entering force to the ions and small electrical ligands. The effect of these chemical (NO) and physical (EMF) factors on the expression of the stemness and neuronal differentiation markers in rat bone marrow mesenchymal stem cells (BMSC) was investigated. The cells were treated with low (50micromolar) and high (1mM) concentrations of Deta-NO as a NO donor molecule and 50Hz low frequency EMF. The expression of pluripotency and neuronal differentiation genes and proteins was investigated using real time qPCR and Immunocytochemistry techniques. The simultaneous treatment of EMF with NO (1mM) led to the down-regulation of stemness markers expression and up-regulation of neuronal differentiation markers expression. Cell proliferation decreased and cell morphology changed which caused the majority of cells obtains neuronal protein markers in their cytoplasm. The decrease in the expression of neuronal differentiation Nestin and DCX markers without any change in the expression of pluripotency Oct4 marker (treated with low concentration of NO) indicates protection of stemness state in these cells. Treatment with NO demonstrated a double behavior. NO low concentration helped the cells protect the stemness state but NO high concentration plus EMF pushed cells into differentiation pathway.

Association of expression of selenoprotein P in mRNA and protein levels with metabolic syndrome in subjects with cardiovascular disease: Results of the Selenegene study.

BACKGROUND: Selenoprotein P (SeP) is involved in transporting selenium from the liver to target tissues. Because SeP confers protection against disease by reducing chronic oxidative stress, the present study aimed to assess the level of SeP in the serum of patients with metabolic syndrome (MetS) with a history of cardiovascular disease (CVD). METHODS: A cross-sectional study was conducted in 63 and 71 subjects with and without MetS in the presence of documented CVD. All demographic, anthropometric and cardiometabolic variables (lipids, blood glucose, blood pressure) were assessed. Lifestyle-related factors and personal history and familial CVD risk factors were recorded. The expression of SELP in mRNA and protein levels in the serum was measured, and MetS was determined using ATPIII criteria. Binary logistic regression analysis demonstrated MetS and SeP to be dependent and independent variables, respectively. RESULTS: Mean of systolic and diastolic blood pressure, triglyceride, high-density lipoprotein-cholesterol, fasting blood sugar, body mass index and waist circumference were higher among subjects with MetS (p = 0.05). The mean of selenium was higher among subjects with MetS, whereas the mean of SeP was lower among subjects with MetS (p < 0.001). In the unadjusted model, the SeP had decreased odds for MetS [odds ratio (OR) = 0.995; 95% confidence interval (CI) = 0.989-1.00] (p < 0.04). Furthermore, the association between MetS and SeP levels remained marginally significant even after adjusting for potential confounders such as age, gender, family history, smoking status and nutrition. SeP and waist circumference show a significant relationship (OR =0.995; 95% CI = 0.990-1.00) (p < 0.033). CONCLUSIONS: We have demonstrated a significant decrease in circulating SeP levels according to MetS status in patients with documented cardiovascular disease.

In vitro polyploidy induction: changes in morphology, podophyllotoxin biosynthesis, and expression of the related genes in Linum album (Linaceae).

MAIN CONCLUSION: Induction of tetraploidy was performed and podophyllotoxin production increased by upregulating the expression level and enzyme activity of genes related to its biosynthesis in tetraploid compared to diploid Linum album. Linum album is a valuable medicinal plant that produces antiviral and anticancer compounds including podophyllotoxin (PTOX). To achieve homogeneous materials, in vitro diploid clones were established, and their nodal segments were exposed to different concentrations and durations of colchicine. This resulted in successful tetraploidy induction, confirmed by flow cytometry, and is being reported for the first time. The highest efficiency of tetraploid induction (22%) was achieved after 72 h exposure to 2.5-mM colchicine treatment. The stable tetraploids were produced after being subcultured three times, and their ploidy stability was confirmed after each subculture. The effects of autopolyploidy were measured on the morphological and phytochemical characteristics, as well as enzyme activity and the expression levels of some key genes involved in the PTOX biosynthetic pathway, including phenylalanine ammonia-lyase (PAL), cinnamoyl-Coa reductase (CCR), cinnamyl-alcohol dehydrogenase (CAD), and pinoresinol-lariciresinol reductase (PLR). The tetraploid plants had larger leaves and stomata (length and width) and lower density stomata. Increasing the ploidy level from diploid to tetraploid resulted in 1.39- and 1.23-fold enhancement of PTOX production, respectively, in the leaves and stem. The increase in PTOX content was associated with upregulated activities of some enzymes studied related to its biosynthetic pathway and the expression of the corresponding genes. The expression of the PAL gene and PLR enzymatic activity had the most positive correlation with the ploidy level in both leaf and stem tissues. Our results verified that autotetraploid induction is a useful breeding method, remarkably increasing the PTOX content in the leaves and stem of L. album.

Prevalence of resistant associated variants (RAVs) in the naïve HCV patient candidate for direct acting antiviral (DAA) therapy.

BACKGROUND: Viral hepatitis C is an important global health problem that affects about 2.2% of humans. Strategies on the control of this hepatotropic virus focused on chemotherapy and surveillance of emerging HCV drug resistant mutants, respectively. HCV genotype 1 response to therapy is one of major interests. The aim of this research was to study the prevalence of resistant associated variants (RAVs) in the naïve HCV patient candidate for direct acting antiviral (DAA) therapy. METHODS: A total of 70 HCV confirmed patients which referred to hospitals affiliated to Iran University of Medial Sciences, Tehran, Iran from May 2014 to March 2015 were enrolled in this cross sectional study. After RNA extraction, RFLP-RT-Nested-PCR was performed for HCV genotyping, then some genotypes 1 and 3 strains were used for further amplification of NS5B gene S282T mutation site and purified products were sequenced. Bioinformatics software was used for analysis of sequences. RESULTS: From a total of 70 HCV patients, 54 were male (mean age (y)±SD 35.1 ± 8.2) and 16 were female (mean age (y)±SD 43.4 ± 10.1); 26 isolates from 1a, 1b and 3a showed that there were no S282T resistant mutants. Moreover, 2 (4.8%) had a synonymous point mutation (C to T). Statistical analysis didn't found any significant correlation between age, sex and genotype variables. CONCLUSION: Finally, it can be concluded that there were no resistant mutants in our HCV genotypes 1 and 3 infected patients and broader scale of studies are required in this area using larger specimens, genotype groups and stages of treatment.