Research in Molecular Medicine
Volume:8 Issue: 1, Feb 2020

Multiple Sclerosis (MS) is the most common chronic inflammatory disease of the white matter of the Central Nervous System (CNS). Proinflammatory cytokine network such as interleukin-6 (IL-6) has a role in the initiation of a destructive immune response in CNS and the progression of the disease. This study aimed to evaluate the comparative levels of IL-6 and IL-8 in MS patients and healthy controls and also to assess any fluctuation in IL-6 and IL-8 levels in MS progression.

This case-control study recruited a total of 183 subjects, including 103 MS patients and 80 sex- and age-matched healthy controls. MS patients included 51 Relapsing Remitting MS (RRMS), 25 secondary progressive MS (SPMS), 27 Primary Progressive MS (PPMS), and Progressive Relapsing MS (PRMS). Clinical findings were collected, and serum levels of IL-6, IL-8, and 25-hydroxyl vitamin D3 (25(OH)D3) were determined with ELISA.

The mean serum level of IL-6 was significantly higher in MS patients than healthy controls (23.8±2.1 vs. 15.6±2.7 pg/mL, P=0.043). MS patients with SPMS had more prominent IL-6 levels than RRMS or PPMS (P=0.008). However, IL-8 levels did not show a significant change either in the patients compared with the controls or in the different forms of MS. MS was more prone to progressive form in male patients. Mean 25(OH)D3 level was significantly lower in MS patients than the controls. Meanwhile, the mean 25(OH)D3 level was surprisingly higher in MS patients with SPMS.

The increased serum level of IL-6 in more advanced MS status, SPMS, suggests that IL-6 but not IL-8 might be a prognostic marker for the disease deterioration. Besides, the tendency for MS to progress to worse stages in affected men is an important finding that needs further clarification.

The production of recombinant proteins in Escherichia coli is one of the most valuable achievements in biotechnology, with many therapeutic and diagnostic applications; however, the aggregation and misfolding of proteins that result in the formation of insoluble inclusion bodies is a disruptive factor in this process. Various solubilization and refolding methods can be used to improve recombinant protein conformation. In this study, we applied a dilution method with a refolding buffer to produce a native form of soluble immature mouse TGF-β1.

The TGF-β1 cDNA which encodes the protein without the signal peptide, was cloned into the pET21-b (+) vector. The target protein was expressed by the transformation of E. coli BL21 cells with the plasmid. The resulting inclusion bodies were solubilized and then diluted in the refolding buffer to make a protein with native structure.

Following PCR of the recombinant plasmid with T7 primers, electrophoresis and sequencing of the amplified product indicated 100% identity of the target sequence with the murine TGF-β1 gene. Finally, the protein solubility and immuno-reactivity were confirmed a 44 kDa protein which conducted with the anti-TGF-β1-specific polyclonal antibody on a western blot.

Our dilution method and refolding buffer effectively converted aggregated immature mouse TGF-β1 to a soluble and immuno-reactive form.

Deregulation of FOXO3a gene which belongs to Forkhead box O (FOXO) transcription factors, can cause cancer (e.g. breast cancer). FOXO factors have important role in ubiquitination, acetylation, de-acetylation, protein-protein interactions and phosphorylation. Understanding the regulation and mechanisms of FOXO3a can lead to cancer treatment. The aim of this study recent association of data mining with genetics has provided a strong tool for knowledge discovery.

Using genetics and bioinformatics, 30 sequences of FOXO3a genes were extracted from different species and were used in two datasets including 65 nucleotide features and 51 tandem repeat sequences. Then, we used different feature weighting and decision tree data mining algorithms on these datasets.

Among nucleotide features, the frequency of AA dinucleotide was the most important genomic feature for FOXO3a gene identification. Among tandem repeat sequences, the strings of TTTTTTTTT, GAGGAGGAG, CGGCGGCGGCGG and CGGCGGCGGCGGCGG were the most effective ones to distinguish FOXO3A gene between different species.

The results of this study are important in understanding FOXO3a gene and developing a pathway for cancer and gene therapies in humans.

 Human coagulation factor IX (hFIX) is a glycoprotein with two N-glycosylation sites at the activation peptide. Since the activation peptide is removed in mature hFIX, the exact role of N-glycosylation is unclear. To investigate the role of N-glycosylation in the secretion and activity of hFIX, we inhibited N-glycosylation by tunicamycin in the stable Human Embryonic Kidney (HEK)- coagulation Factor IX (FIX) cells.

 After the treatment of stable FIX-expressing HEK cells in the presence or absence of tunicamycin, the expression and activity of the recombinant FIX (rFIX) were determined in culture medium and cell lysate with enzyme-linked immunosorbent assay and clotting test, respectively.

Based on the data analysis, total concentrations of FIX in stable HEK-FIX was the same in the media with and without tunicamycin. But throughout the post-induction period, the intracellular and secreted levels of FIX in tunicamycin-treated HEK-FIX cells increased and decreased, respectively, compared with those of control HEK-FIX cells, though the results were not significant. These results indicate that disrupting the synthetic process may slightly reduce the FIX levels secreted in HEK-FIX cells.

 Although glycosylation plays a vital role in the folding and secretion of the proteins, it does not affect the secretion of FIX. Besides, the N-glycosylation of the produced FIX failed to play a significant role in its activity.

Paraquat (PQ), as the most widely used herbicide in agriculture, induces poisoning in humans and animals mainly through oxidative stress. This study aimed to investigate the protective effects of nano-curcumin compared with curcumin against PQ-induced mitochondrial dysfunction.

Thirty-six Wistar rats were used in this study. They were separated into 6 groups: control subjects and animals poisoned with PQ that received treatment with or without curcumin and nano-curcumin for 7 days. Liver mitochondria were isolated, and oxidative stress markers, including Total Antioxidant Capacity (TAC), Lipid Peroxidation (LPO), Catalase (CAT) and Superoxide Dismutase (SOD) activity, as well as viability and mitochondrial membrane potential, were assessed.

Poisoning by PQ significantly increased the LPO levels and considerably decreased the TAC, CAT, and SOD activity compared with the control subjects (P<0.05). PQ-induced oxidative significantly damaged the viability and membrane potential of mitochondrial compared with controls (P<0.05). Administration of nano-curcumin significantly increased the SOD activity, as well as viability and mitochondrial membrane potential in the PQ group (P<0.05). Besides, treatment by nano-curcumin in the PQ group significantly improved the lipid peroxidation process (P<0.05).

Nanocurcumin is more effective than curcumin in reducing the PQ-induced oxidative damage

Pulmonary disorders caused by parasites are common in many tropical regions. Acanthamoeba is an opportunistic parasite, and most of its dangerous complications are seen in patients with immune deficiencies. Considering the high dissemination of Acanthamoeba parasite in water, soil, and fine airborne dust in Iran, this research was performed to study the rate of pulmonary secretions infection to Acanthamoeba in patients with immune system disorder.

This cross-sectional study was done in one year (2017 to 2018) in Arak City. The study sample was selected from immunodeficient subjects who had chronic obstructive pulmonary disease, too. The demographic data were also collected from all study samples. The respiratory sample was obtained from each selected patient by Bronchoalveolar Lavage (BAL). Each sample was examined by smear staining, cultivation, and molecular methods.

Out of 64 immunocompromised patients investigated in the current research, the Acanthamoeba infection was found in 100%, 98.4%, and 0% of their BAL with culture, molecular, and direct methods, respectively.

The outcomes of our research indicated that selecting the diagnostic method in agreement with the kind of sample has a remarkable role in recognizing the contamination. The results of this research showed that the direct microscopy test of Giemsa stained smear was not suitable for detecting this kind of parasitic infection in the BAL sample. Therefore, awareness of the occurrence of Acanthamoebiasis in immunocompromised subjects is essential for preventing the dangerous complications of this parasite.