Reports of Biochemistry and Molecular Biology最新文献

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: 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: 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.

Background: Normally happening substances like flavonoids are regarded as active candidates for the treatment and prevention of cancer The purpose of this study was to see how Iraqi E. arvense total flavonoid affected cell lines biologically and human lung fibroblast normal cell line (WISH).

Methods: Plant powder was extracted by reflex apparatus, then thin-layer chromatography (TLC) was used to determine total flavonoids. Cytotoxicity assay (MTT) was used to determine the cytotoxic activity of the prepared plant against human breast cancer (MCF-7), cells human cervix cancer (HELA), human colon cancer (Caco-2) and human lung fibroblast normal cell line (WISH).

Results: The flavonoids Rutin, Quercetin, Kaempferol, and luteolin were detected using the Thin Layer Chromatography (TLC) technique. In contrast to the negative control, the extract inhibited cell growth to a highest of 82.158% for MCF-7 and 61.360% for Caco-2 at the concentration (100 µg/ml), and (54.880%) for Hela cell line at the concentration (100 µg/ml). In addition, the concentration (6.25 µg/ml) of total flavonoid extract produced a decrease in the growth of the normal WISH cell line to reach (1.094%).

Conclusion: Equisetum arvense contain high amounts of flavonoids, the qualification of some flavonoids compounds was detected using TLC. The total flavonoids showed significant cytotoxic activity against various types of cancer cell lines and normal cell line in vitro, the antitumor activity was highly efficient in a dose and cell type dependent manner.

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease with inflammatory nature. One of the leading causes of death in SLE patients is cardiovascular (CVS) morbidity. MiRNA-24 is highly expressed in vascular endothelial cells (VECs). This dysregulated expression pattern is associated with dysfunction or even damage of VECs and leads to the occurrence of cardiovascular diseases. YKL- 40 is an inflammatory glycoprotein involved in the pathogenesis of endothelial dysfunction and thereby atherosclerosis. In this work, we aimed at illustrating the possible role of miR-24 and its target YKL-40 in the pathogenesis of the CVS morbidity associated with SLE.

Methods: This work was conducted on 40 SLE patients and 40 healthy controls. Quantitative real-time PCR (qPCR) was done to estimate the expression level of miRNA-24 in serum. In addition, we measured the serum level of YKL-40 using ELISA.

Results: miR-24-fold change was found to be down-regulated, whereas serum YKL- 40 was up-regulated among SLE patients with observed significant and negative correlation between the two parameters.

Conclusion: Our study provided an insight about the role of miR-24 and its target serum YKL-40 protein in the development of SLE-related inflammation and atherosclerosis.

Background: Diabetic retinopathy (DR) is one of diabetes mellitus complication and occurred in retinal microvascular. This study was aimed to investigate the efficacy of Sambiloto, Andrographis paniculate (A. paniculata) extract on glycemic profile, antioxidant and inflammatory cytokine parameters in diabetic rats, and phytochemical analysis of A. paniculata.

Methods: A. paniculata extract (APE) was carried out by maceration with ethanol. Diabetes mellitus in Wistar male rats was induced with streptozotocin. Retinal vessel diameters were estimated using a method by Vucetic. Inflammatory cytokine and antioxidant parameters were evaluated in retinal tissue. The alkaloid and flavonoid contents in extract were analyzed using thin layer chromatography method.

Results: Funduscopic examination presented some changes in the diameter of the blood vessels. The vessel diameter in the diabetic retinopathy group with APE in concentration of 100 and 200 mg/kg BW groups was significantly lower than in the DR group (p<0.05). The administration of APE in dosages of 100 and 200 mg/kg BW showed reduced glutathione, SOD, and catalase levels compared to the DR group (p<0.05).

Conclusion: A. paniculata extract doses of 100 and 200 mg/kg BW improved diabetic retinopathy in rats through hypoglycemic effects, antioxidant effects, and anti-inflammatory mechanisms.

Background: Immunological alterations in schizophrenic patients have been considered during last decade. There are no remarkable reports on the changes of IL-17A and IL-21 in schizophrenic patients. Therefore, the purpose of this study was to evaluate changes of serum IL-17A and IL-21 in schizophrenic patients in comparison with healthy controls.

Methods: In the present study serum levels of IL-17A and IL-21 in 30 patients with schizophrenia before treatment and three months after treatment were measured by enzyme-linked immunosorbent assay (ELISA) and compare to 30 match healthy control group.

Results: Serum levels of IL-21 in schizophrenic patient was significantly higher than control group (P= 0.001). Serum levels of IL-17A in the schizophrenic patients had no significant changes than the control group (P= 0.4). Serum levels of IL-17A in patients with schizophrenia three months after treatment than before treatment had no significant change (P=0.7) and IL-21 serum levels in schizophrenic patient three month after treatment was not significant changed in comparison with this group before treatment (P= 0.06).

Conclusion: The serum levels of interlukine-21 is elevated in schizophrenic. Results of this study showed that IL-21 might be involved in the pathologic mechanism of schizophrenia.

Background: COVID-19 is caused by the Severe Acute Respiratory Distress Syndrome Coronavirus 2. Since the antioxidant mechanisms such as glutathione peroxidase or superoxide dismutase are downregulated during infection by the virus, there is an imbalance in the oxidant-antioxidant system. In this study we aimed to identify the effect of COVID-19 on the antioxidant defense mechanism by comparing the concentrations of antioxidants and microminerals in COVID-19 patients and healthy controls.

Methods: This cross-sectional analytical study involved 200 patients at Kasturba Hospital, Manipal University. The serum concentrations of antioxidants and minerals were determined to establish the impact of COVID-19 on antioxidants mechanism and nutrient status in COVID-19 patients.

Results: The serum concentrations of GPX (10.36 ± 2.70 ≥ 5.82 ± 1.64 mKAT/L, p < 0.0001) and copper (2192.5 ± 449.8 ≥ 782.15 ± 106.5 µg/dL, p < 0.0001) were significantly greater, and zinc (34.78 ± 4.5 ≤ 81.07 ± 10.13 µg/dL, p < 0.0001) was significantly less, in the study group than in controls. The Pearson correlation between serum SOD and zinc was significant (r = 0.491, p < 0.0001) indicating the importance of zinc in maintaining and improving SOD activity. No significant correlations were observed between copper and SOD (r = -0.089) or iron and CAT (r = -0.027).

Conclusion: Our study demonstrated the expected increase in oxidant-radical production during COVID-19 by estimating the altered concentrations of antioxidants and the minerals required to neutralize the elevated ROS. This finding is not novel but adds to the existing literature, which recommends nutritional supplementation of microminerals and antioxidants.