Reports of Biochemistry and Molecular Biology最新文献

Background: Periodontal disease is an inflammatory condition affecting the tooth's supporting tissues, resulting in gradual loss of periodontal ligament (PDL), alveolar bone, and gum resorption. Neutrophil and monocyte/macrophage, destructive proteases like matrix metalloproteinase (MMP)-3 and MMP-9 play pivotal roles in such lesions in periodontitis. Therefore, this study aims to compare the level of MMP-3 and MMP-9 gene expression in patients with or without periodontitis in an Iranian population.

Methods: This cross-sectional study was carried out on 22 chronic periodontitis patients and 17 healthy control subjects referred to the department of periodontology, Mashhad Dental School. In both groups, the gingival tissue was removed during surgery and transferred to the Molecular Biology Laboratory for MMP-3 and MMP-9 gene expression evaluation. The qRT-PCR, TaqMan method was used for gene expression assessments.

Results: The average age of periodontitis patients was 33± 5 years, and in controls, 34.7± 6 with no significant differences. The mean MMP-3 expression in periodontitis patients was 146.67±38.7, and in controls, 63.4±9.1. The difference was statistically significant (P=0.04). The mean expression of MMP-9 in periodontitis patients and controls were 103.8± 21.66 and 87.57± 16.05, respectively. Although the target gene expression in patients was higher, the difference was insignificant. Furthermore, there was not any significant correlation between age or gender with the expression of MMP3 or MMP9.

Conclusion: The study demonstrated that the MMP3 seems to have a destructive impact on the gingival tissue in chronic periodontitis, but not MMP9.

Background: Double-stranded fragmented extracellular DNA is a participant, inducer, and indicator of various processes occurring in the organism. When investigating the properties of extracellular DNA, the question regarding the specificity of exposure to DNA from different sources has always been raised. The aim of this study was to perform comparative assessment of biological properties of double-stranded DNA obtained from the human placenta, porcine placenta and salmon sperm.

Methods: The intensity of leukocyte-stimulating effect of different dsDNA was assessed in mice after cyclophosphamide-induced cytoreduction. The stimulatory effect of different dsDNA on maturation and functions of human dendritic cells and the intensity of cytokine production by human whole blood cells was analyzed ex vivo. The oxidation level of the dsDNA was also compared.

Results: Human placental DNA exhibited the strongest leukocyte-stimulating effect. DNA extracted from human and porcine placenta exhibited similar stimulatory action on maturation of dendritic cells, allostimulatory capacity, and ability of dendritic cells to induce generation of cytotoxic CD8+CD107a+ T cells in the mixed leukocyte reaction. DNA extracted from salmon sperm stimulated the maturation of dendritic cells, while having no effect on their allostimulatory capacity. DNA extracted from human and porcine placenta was shown to exhibit a stimulatory effect on cytokine secretion by human whole blood cells. The observed differences between the DNA preparations can be caused by the total methylation level and are not related to differences in oxidation level of DNA molecules.

Conclusions: Human placental DNA exhibited the maximum combination of all biological effects.

Background: Exosomes are nanoscale vesicles widely used as drug delivery systems. Mesenchymal stem cell (MSC)-derived exosomes have shown immunomodulatory potential. This study optimized loading OVA into the mice adipose tissue-derived MSC-isolated exosomes to prepare the OVA-MSC-exosome complex for allergen-specific immunotherapy.

Methods: MSCs were harvested from mice adipose tissue and characterized by flow cytometry and evaluating differentiation potential. The exosomes were isolated and characterized via Dynamic Light Scattering, Scanning Electron Microscopy, and flow cytometry. Different concentrations of ovalbumin were incubated with MSC-exosome in various durations to optimize a more suitable protocol. BCA and HPLC analysis were used to quantify, and DLS was applied to qualify the prepared formulation of the OVA-exosome complex.

Results: The harvested MSCs and isolated exosomes were characterized. Analysis of the OVA-exosome complex revealed that OVA in primary 500 μg/ml concentration and incubation for 6 h results in higher efficacy.

Conclusions: Loading OVA into MSC-derived exosomes was successfully optimized and could be administrated for allergen-specific immunotherapy in the animal model.

Background: Caffeine is generally suggested to increase VO2max in endurance performance. Nevertheless, the response to caffeine ingestion does not seem to be uniform across individuals. Therefore, caffeine ingested timing on endurance performance based on the type of CYP1A2 single nucleotide polymorphism rs762551, that were classified as fast and slow metabolizers, need to be evaluated.

Methods: Thirty participants participated in this study. DNA was obtained from saliva samples and genotyped using polymerase chain reaction-restriction fragment length polymorphism. Each respondent completed beep tests under three treatments blindly: placebo, 4 mg/kg body mass of caffeine one hour, and two hours before test.

Results: Caffeine increased estimated VO2max in fast metabolizers (caffeine=29.39±4.79, placebo=27.33±4.02, p<0.05) and slow metabolizers (caffeine=31.25±6.19, placebo=29.17±5.32, p<0.05) in one hour before test. Caffeine also increased estimated VO2max in fast metabolizers (caffeine=28.91±4.65, placebo=27.33±4.02, p<0.05) and slow metabolizers (caffeine=32.53±6.68, placebo=29.17±5.32, p<0.05) in two hour before test. However, for slow metabolizers, the increasing was greater when caffeine was administered two hours before test (slow=3.37±2.07, fast=1.57±1.62, p<0.05).

Conclusion: Genetic variance may affect the optimal caffeine ingestion timing, sedentary individuals who want to enhance their endurance performance may ingest caffeine 1 hour before exercise for fast metabolizers and 2 hours before exercise for slow metabolizers.

Background: : Cancer continues worldwide. It has been reported that OTUB1, a cysteine protease, plays a critical role in a variety of tumors and is strongly related to tumor proliferation, migration, and clinical prognosis by its functions on deubiquitination. Drug advances continue against new therapeutic targets. In this study we used OTUB1 to develop a specific pharmacological treatment to regulate deubiquitination by OTUB1. The aim of this research is to regulate OTUB1 functions.

Methods: By molecular docking in a specific potential OTUB1 interaction site between Asp88, Cys91, and His26 amino acids, using a chemical library of over 500,000 compounds, we selected potential inhibitors of the OTUB1 catalytic site.

Results: Ten compounds (OT1 - OT10) were selected by molecular docking to develop a new anti-cancer drug to decrease OTUB1 functions in cancer processes.

Conclusion: OT1 - OT10 compounds could be interacting in the potential site between Asp88, Cys91, and His265 amino acids in OTUB1. This site is necessary for the deubiquitinating function of OTUB1. Therefore, this study shows another way to attack cancer.

Background: Macrophages are essential cellular components in various body tissues and tumor microenvironments. The high infiltration of macrophages into the tumor microenvironment determines the importance of ex vivo treatment of personalized macrophages with recombinant cytotoxic T-lymphocyte-associated protein 4 (rCTLA-4), programmed death-ligand 1 (rPD-L1), and programmed cell death protein 1 (rPD-1) proteins to block immune checkpoints.

Methods: We investigated the development of humoral immunity against CTLA-4, PD-L1, and PD-1 receptors by introducing macrophages treated ex vivo with the corresponding proteins into mice. Peritoneal macrophages from BALB/c mice were cultured in medium containing recombinant human CTLA-4, PD-L1, and PD-1 proteins. Macrophages processing recombinant proteins were analyzed via immunofluorescence staining using antibodies against CTLA-4, PD-L1, and PD-1. The treated macrophages were administered intraperitoneally to mice to induce anti-CTLA-4, anti-PD-L1, and anti-PD-1 antibodies. The antibody titer in vaccinated mice was determined via enzyme-linked immunosorbent assays, followed by statistical analysis of the results. The specificity of the antibodies was determined via immunofluorescence staining in MCF7 cells.

Results: The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 induced the formation of specific antibodies in vaccinated mice. The various rPD-L1 and rPD-1 concentrations used to treat macrophages had no significant effect on the specific antibody titers, while the anti-rCTLA-4 titer was dependent on the protein concentration in the culture medium. Immunofluorescence analysis revealed that anti-CTLA-4 and PD-L1 antibodies reacted with MCF7 cells.

Conclusion: The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 can help induce humoral immunity and develop new approaches for cancer immunotherapy.

Background: IgA is widely used as Upper Respiratory Tract Infection (URTI) risk marker, as a lower concentration in sIgA indicates a higher incidence of URTI. This study aimed to investigate the effect of different types of exercise; combined with Tempeh consumption in increasing sIgA concentration in saliva sample.

Methods: 19 sedentary male subjects aged 20-23 were recruited and assigned into 2 groups based on the exercise type, endurance (n=9), and resistance (n=10). These subjects underwent 2 weeks of Tofu and Tempeh consumption, then were assigned to do exercises based on their groups.

Results: This study showed an increased mean value of sIgA concentrations in the endurance group; the baseline value, after food treatment, and after food and exercise treatment were 71.726 ng/mL, 73.266 ng/mL, and 73.921 ng/mL, respectively for Tofu treatment; and 71.726 ng/mL, 73.723 ng/mL, and 75.075 ng/mL, respectively for Tempeh treatment. While in the resistance group, there was also an increase in the mean value of sIgA concentrations; baseline, after food treatment, and after food and exercise treatments were 70.123 ng/mL, 71.801 ng/mL, and 74.430 ng/mL, respectively for Tofu treatment; and 70.123 ng/mL, 72.397 ng/mL, and 77.216 ng/mL, respectively for Tempeh treatment. These results indicated that combining both Tempeh consumption and moderate intensity resistance exercise was more effective to increase sIgA concentration.

Conclusion: This study showed that combining moderate intensity resistance exercise with consumption of 200 gr Tempeh for 2 weeks was more effective in increasing sIgA concentration; compared to endurance exercise and Tofu consumption.

Background: The role of the basic fibroblast growth factor (bFGF) has well known in the angiogenesis and ulcer healing. In this study, we aimed to evaluate the effects of bFGF on tissue repair in a rat oral mucosal wound.

Methods: Musosal wound induced on the lip mucosa of rats and bFGF was injected along the edge of the mucosal defect immediately after surgery. The tissues were collected on days 3, 7 and 14 after the wound induction. The micro vessel density (MVD) and CD34 expression were done by histochemical studies.

Results: The bFGF significantly accelerated granulation tissue formation and MVD was increased three days after ulcer induction but decreased 14 days after surgery. The MVD was significantly higher in the bFGF-treated group. The wound area was decreased in all groups time-dependently and a statistically significant difference (p value?) was observed between the bFGF-treated group and untreated group. The wound area was smaller in the bFGF-treated group compared to the untreated group.

Conclusions: Our data demonstrated that bFGF can accelerated and facilitated wound healing.

Background: Pediatric immune thrombocytopenic purpura (ITP) is an autoimmune disease; whose etiology is unknown. lncRNAs are regulators of numerous actions, which participate in the development of autoimmune diseases. We evaluated the expression ofNEAT1 and Lnc-RNA in dendritic cell (Lnc-DC) in pediatric ITP.

Methods: Sixty ITP patients and 60 healthy subjects were enrolled in the present study; Real-time PCR was performed to assess the expression levels of NEAT1 and Lnc-DC in sera of children with ITP as well as healthy children.

Results: Both lncRNAs, NEAT1 and Lnc-DC were significantly upregulated in ITP patients in comparison to controls (p <0.0001 and P= 0.001 respectively). Furthermore, significant upregulation of the expression levels of NEAT1 and Lnc-DC were observed in the non-chronic compared with chronic ITP patients. Also, there was significant negative correlation between each of NEAT1 and Lnc-DC and platelet counts before treatment (r= -0.38; P= 0.003 and r= -0.461; P< 0.0001, respectively).

Conclusions: serum lncRNAs, NEAT1 and Lnc-DC could be used as potential biomarkers in differentiating childhood ITP patients and healthy controls in addition to differentiating non-chronic from chronic ITP which may provide a theoretical basis for the mechanism and treatment of immune thrombocytopenia.