Integrative analysis of OIP5-AS1/HUR1 to discover new potential biomarkers and therapeutic targets in multiple sclerosis

Multiple sclerosis (MS) is a devastating autoimmune disease of the central nervous system associated with demyelination and axonal injury. This study was designed to find potential lncRNAs and their targets that are associated with the molecular basis of MS pathogenesis. In this study, peripheral blood samples were obtained from 50 relapsing-remitting MS (RR-MS) patients and 50 healthy controls. lncRNAs and their target were selected for validation using TaqMan Real-Time PCR. Interactions were studied based on approaches that used to investigation biological functions and signaling pathways affected by differentially expressed messenger RNAs (mRNAs). The results of this study indicate an increase in the expression of HUR1 (p = 0.0001), CPSF7 (p = 0.02), and reduction of CSTF2 expression (p = 0.04). Also, an increase in the expression of OIP5-AS1 (p = 0.01) was observed in men less than 30 years old. We performed a comparative analysis of the long noncoding RNAs (lncRNAs), and then we ranked them as candidate biomarkers according to a decreasing area under the receiver operating characteristic (ROC) curve (AUC) and plotted the results. Dysregulation of lncRNA expression has been linked to diseases. Further studies on the HUR1 gene can be used as diagnostic tools for the identification of high-risk individuals in families with a history of disease before, during, and even after treatment. Our data uncovered the expression profiles of lncRNAs and mRNAs in MS patients, which will help delineate the molecular mechanisms in MS pathogenesis. However, further studies need to determine the precise role of these genes in the pathological process in MS.     

Genetic discoveries and advances in late-onset Alzheimer’s disease

Alzheimer’s disease (AD) is a heterogeneous disorder with multiple patterns of clinical manifestations. Recently, due to the advance of linkage studies, next-generation sequencing and genome-wide association studies, a large number of putative risk genes for AD have been identified using acquired genome mega data. The genetic association between three causal genes, including amyloid precursor protein, presenilin1, and presenilin2 in early-onset AD (EOAD), was discovered over the past few decades. These discoveries showed that there should be additional genetic risk factors for both EOAD and late-onset AD (LOAD) to help fully explain the leading molecular mechanisms in a single pathophysiological entity. This study reviews the clinical features and genetic etiology of LOAD and discusses a variety of AD-mediated genes that are involved in cholesterol and lipid metabolism, endocytosis, and immune response according to their mutations for more efficient selection of functional candidate genes for LOAD. New mechanisms and pathways have been identified as a result.     

Do Fertilizers and Irrigation Disruption Change Some Physiological Traits of Safflower?

To investigate the effects of nanofertilizers and biofertilizers on the morpho-physiological and biochemical traits of safflower under full irrigation and water deficit stress, this study was carried out as a split-plot experiment based on a Randomized Complete Block Design with three replications at Urmia University in 2015. The main plot was full irrigation (control) and irrigation disruption at heading, flowering, and grain filling stages. Fertilizers, including control (without fertilizer), biofertilizer, water spray, foliar application of nanofertilizers, chemical fertilizers, and combined application of fertilizers, were assigned to the subplot. Plants under full irrigation and combined fertilizers had maximum height and chlorophyll a, whereas the lowest ones were obtained in irrigation disruption at the heading stage and control treatments. The maximum oil content (28.41%) was detected in irrigation disruption at the grain filling stage and nanofertilizer treatment, the lowest (21.96%) was obtained at irrigation disruption at the flowering stage and water spray treatment. The highest proline (397.21 µg g⁻¹ fresh leaf) was found in irrigation disruption at the grain filling stage and water spray treatment, and the lowest (154.68 µg g⁻¹ fresh leaf) was obtained at full irrigation and water spray treatment. Irrigation disruption at the heading stage and control treatments decreased carbohydrate content of fresh leaves by 86.54% compared to full irrigation and the combined fertilizers treatment. Irrigation disruption increases saturated fatty acids (palmitic and stearic acid) and decreases vitamin E and linoleic acid. The combined application of fertilizers significantly increased safflower oil quality. Overall, concerning the obtained highest oil percentage (28.41%), irrigation disruption during grain filling reduced water consumption and application of combined fertilizer via improving oil quality, so it is recommended to farmers.

     

Isolation and characterization of a novel scFv antibody fragments specific for Hsp70 as a tumor biomarker

Many studies have shown that more than 50% of tumors express heat shock protein 70 kDa (Hsp70) at the plasma membrane surface while not seen in normal cells, therefore it is a promising therapeutic target in human cancers. Hence, we used phage display technology to produce a single-chain fragment variable (scFv) antibody against human Hsp70. For this, a target peptide from human Hsp70 was designed using bioinformatics studies and was chemically synthesized. Then, the selection was performed using four rounds of biopanning with a stepwise decreased amount of the target peptide. Fourteen positive scFv clones were selected using monoclonal phage enzyme-linked immunosorbent assay screening, which was further characterized by means of the polymerase chain reaction and DNA sequencing. Among them, the G6 clone was selected to express scFv into the Escherichia coli. Expression and purification of the scFv shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by Western blot analysis. In silico analysis confirmed specific binding of the scFv to Hsp70 in CDR regions. The specificity of the scFv measured by surface plasmon resonance and immunofluorescence of the A549 human lung carcinoma cell line confirmed the in vitro function of the scFv. Based upon these findings, we propose a novel anti-human Hsp70 scFv as potential immunotherapy agents that may be translated into preclinical/clinical applications.     

In vitro inhibition of human erythrocyte hexokinase by various hyperglycemic drugs

Hemolysis is the red blood cell abnormality most often associated with adverse effect of drug therapy. Drug‐induced or drug‐associated hyperglycemia could decrease the activity of hexokinase. The aim of this study was to investigate the inhibitory effects of some commonly used drugs that have hyperglycemic side effect on the human erythrocyte hexokinase enzyme in vitro. Hexokinase was purified from human erythrocytes using sequential chromatography, with a specific activity of 0.96 ± 0.18 U/g hemoglobin, and assayed in the presence of selected drugs that have hyperglycemic side effect. The IC₅₀ were determined from the regression analysis graph. Correlation analysis showed that there was positive correlation between the hyperglycemic side effect of some of the tested drugs and decrease of hexokinase activity. This suggests that, at least in part, these drugs exert their hyperglycemic effect by inhibiting glucose phosphorylation by the hexokinase, which consequently causes the glucose accumulation.     

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Anticancer role of oxindole compounds is well documented. Here, we synthesized new derivatives of 3-hydroxy-2-oxindole functionalized at position 3 (1a–f) which are expected to have antiproliferative activity in cancer cells. Human prostate cancer cell line (DU145) was treated with the synthesized derivatives at 40-μM concentration for 24, 48, and 72 h. Compounds 1-ethyl-3-hydroxy-1,1′,3,3′-tetrahydro-2H,2′H-3,3′-biindole-2,2′-dione (1d), 5-bromo-1-ethyl-3-hydroxy-1,1′,3,3′-2H,2′H-3,3′-biindole-2,2′-dione (1e), and 5-chloro-1-ethyl-3-hydroxy-1,1′,3,3′-tetrahydro-2H,2′H-3,3′-biindole-2,2′-dione (1f) were found to significantly reduce DU145 cell viability at 48 and 72 h whereas no significant changes were observed up to 24 h. The compounds 1e and 1f showed the most cytotoxicity effect and had a similar antiproliferative activity on DU145 cell line. They have halogen and ethyl substitutions at positions 5 and 1, respectively. The IC₅₀ of compound 1e for DU145 and A375 cells at 48 h was determined. The apoptotic effects and cell cycle progression of compound 1e at 1/2 × IC₅₀ (55 μM) concentration in DU145 cells were investigated by nuclei staining, comet assay, flow cytometry, and scanning electron microscopy (SEM). The results obtained showed that this compound increased the percentage of tail DNA, increased the occurrence of the sub-G1 phase, and induced G2M arrest and apoptosis in DU145 cells after exposure for 48 h to a 55-μM concentration. The SEM images revealed cell contraction at 24 h, cell condensation, plasma membrane blebbing, and formation of apoptotic bodies at 48 and 72 h. These observations suggest that the antiproliferative activity of compound 1e may be to induce apoptosis in DU145 cells.     

Antioxidant status and physiological responses of wheat (Triticum aestivum L.) to cycocel application and bio fertilizers under water limitation condition

In order to study antioxidant status and physiological responses of wheat to cycocel (CCC) and bio fertilizers application under water limitation condition, a factorial experiment was conducted based on randomized complete block design with three replications in 2015. Treatments included water limitation in three levels [normal irrigation (I₁) as control; moderate water limitation (I₂) or irrigation withholding at 50% of heading stage; severe water limitation (I₃) or irrigation withholding at 50% of booting stage]; four bio fertilizer levels [(no bio fertilizer (F₀), seed inoculation by Azotobacter chrocoocum strain 5 (F₁), Pseudomonas putida strain 186 (F₂), Azotobacter?+?Pseudomonas (F₃))] and four CCC levels [(without CCC as control (C₀), application of 400 (C₁), 800 (C₂) and 1200 (C₃) mg/l)]. The results showed that water limitation decreased the chlorophyll-a, chlorophyll-b, total chlorophyll, carotenoid, stomata conductance, leaf area index (LAI) and relative water content of wheat, but activity of catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO) enzymes and proline content were increased. Similar results were observed in CAT, POD and PPO activities due to bio fertilizers and CCC application. Besides the water limitation effects, CCC-treated plants displayed a significant decrease in stomata conductance and LAI. Generally, it was concluded that the application of bio fertilizers and CCC can be a proper tool for increasing wheat yield under water limitation.     

Comparison between single and three portal laparoscopic splenectomy in dogs

ABSTRACT: BACKGROUND: Single incision laparoscopic surgery (SILS) is a newly growing technique to replace a more invasive conventional multiple portal laparoscopic surgery. The objective of this study was to compare single (SILS) with three portal (Conventional) laparoscopic splenectomy in dogs. Mongrel dogs (n = 18), weighting 15 +/- 3 kg, were selected for this study (n = 12 SILS; n = 6 conventional). The area from xiphoid to pubis was prepared under aseptic conditions in dorsal recumbency with the head down and tilted 30 degree in the right lateral position. Pneumoperitoneum was established by CO2 using an automatic high flow pressure until achieving 12 mm Hg. Instrumentation used consisted of curved flexible-tip 5 mm Maryland forceps and ultracision harmonic scalpel for sealing and cutting of the vessels and splenic attachments. RESULTS: All dogs recovered uneventfully. The splenectomy procedure using SILS and conventional methods were significantly different in the respective operative time (29.1 +/- 1.65 vs. 42.0 + 2.69 min) and the length of the surgical scar (51.6 +/- 1.34 mm vs. 72.0 +/- 1.63 mm; P < 0.001). There were no post-operative wound complication including inflammation, infection, hernia formation and dehiscence up to one month after surgery. Meanwhile, the conversion to open surgery or application of additional portals was not required in both approaches. CONCLUSION: This study demonstrated that SILS is a safe and feasible operation and could be used as an alternative approach to three portal (Conventional) for splenectomy in dog.