rasul moukhah
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Background
COVID-19 may trigger the progression of atherosclerosis, potentially leading to its clinical manifestation. Among the serum biomarkers related to atherosclerotic disease, it seems helpful to evaluate biochemical factors such as CD147, ACE2, and ACE in patients with COVID-19.
MethodsIn this case-control study, serum samples from 90 patients were analyzed. The cohort included 30 patients hospitalized in the intensive care unit with COVID-19, 30 patients hospitalized in the ward with COVID-19, and 30 healthy individuals from Ayatollah Kashani Hospital (Tehran, Iran). These patients were admitted between December 10, 2022, and March 10, 2023. The concentrations of cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and the activity levels of angiotensin-converting enzyme (ACE), angiotensin-converting enzyme 2 (ACE2), troponin, CPK, CPK-MB, and CD147 were determined and analyzed among the three groups.
ResultsThere was no significant difference in age between the patients (55.40 ± 10.34 years) and the control group (58.34 ± 11.71 years). SARS-CoV2 viral RNA was detected in the pharyngeal swab samples of the COVID-19 patients. The levels of CD-147 (2528.43 ± 12.43 vs. 2176.7 ± 9.87 vs. 1346.3 ± 14.23), ACE (83 ± 3.05 vs. 97 ± 1.55 vs. 30.1 ± 2.32), troponin (0.972 ± 0.25 vs. 0.784 ± 0.21 vs. 0.021 ± 0.68), and cholesterol (199.73 ± 2.43 vs. 175.87 ± 5.21 vs. 144.97 ± 8.74) were significantly higher in severe cases compared to non-severe cases (P < 0.05). ACE (AUC = 0.894) and CD147 (AUC = 0.821) had the highest sensitivity and specificity among the studied factors, respectively. Patients with normal levels of ACE and CPK-MB had a 0.16 times lower and 4.37 times greater chance of experiencing severe disease, respectively. Three strong correlations were found among the studied parameters: CD147 with ACE2 (r = 0.721) and cholesterol with triglyceride (r = 0.602) (P < 0.05).
ConclusionsExamining ACE and CD147 can help determine a person's susceptibility to atherosclerosis. These two factors have greater sensitivity and specificity and could be used as potential markers for monitoring disease progression.
Keywords: CD-147, ACE, COVID-19, Atherosclerosis, Severity -
Background
Despite global control measures aimed at ending the COVID-19 pandemic, the disease continues to pose a threat to public health. In this study, we examined the serum levels of vitamins C, D, and E, as well as IgG and IgM antibodies in individuals who had previously been vaccinated against COVID-19 and subsequently experienced a relapse of the disease.
ObjectivesThe objective of this study was to investigate the correlation between sufficient levels of vitamins E,D, and C, the severity of the disease, and the immunological response in vaccinated patients who have experienced a recurrence of COVID-19.
MethodsGiven the potential role of vitamins C, D, and E in the management of COVID-19, we conducted a study to examine the serum levels of these vitamins in individuals who had previously been vaccinated against COVID-19 and experienced a disease relapse, characterized by symptoms, such as body pain, shortness of breath, cough, and fever. We compared two groups of hospitalized individuals with varying disease severity to healthy individuals. Additionally, we investigated IgG and IgM antibodies in these patients due to the significance of antibody levels in determining disease severity.
ResultsOur results revealed significant differences in the levels of vitamins C, D, and E between hospitalized individuals and healthy individuals. Furthermore, a notable disparity in serum IgM and IgG levels was observed based on the severity of the disease. However, no significant difference was detected in the average levels of anti-SARS-CoV-2 immunoglobulins among the different groups, whether they had received the AstraZeneca or Sinopharm vaccines.
ConclusionsVitamins C, D, and E play supportive roles in the immune system, aiding the host’s immune response. These findings suggest that maintaining adequate levels of these vitamins may be beneficial in preventing SARS-CoV-2 reinfection and reducing disease severity, particularly in cases where vaccine efficacy is uncertain.
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Background
Bacille Calmette–Guerin (BCG) vaccine is the only vaccine that is used against Mycobacterium tuberculosis, but its efficacy is limited in mycobacterium‑endemic regions. One of the major antigens present on the cell envelope of the vaccine that suppresses the immune system is mannosylated lipoarabinomannan (ManLAM).
Materials and MethodsIn this study, we immunized 4‑week‑old mice with sonicated BCG vaccine injected intraperitoneally two times at an interval of 2 weeks and with ManLAM antigen injected intravenously and then extracted the spleen cells of the immunized mice. They were fused with SP2/0 myeloma cells.
ResultsFive cell line clones producing antibody against ManLAM antigens were prepared and each clone was tested for immunoreactivity against sonicated BCG and purified ManLAM by enzyme‑linked immunosorbent assay (ELISA) and immunoblotting. The clones designated H13F33E11 and H23D91G4 reacted strongly with ManLAM. Immunoblotting using monoclonal antibodies (MAbs) H13F33E11 and H23D91G4 showed that these MAbs bind to ManLAM with a molecular weight of 35 kDa.
ConclusionsIn this study, we produced a monoclonal antibody of immunoglobulin G3 (IgG3) subclass. This MAb can be used for purification of ManLAM in culture media and detection of the antigen in patient’s urine and for increasing the efficacy of BCG vaccine.
Keywords: BCG vaccine, lipoarabinomannan, ManLAM, mannosylated lipoarabinomannan -
In Vitro Evaluation of Inhibitory Effect of Artemisinin and Dihydroartemisinin on Calcineurin EnzymeBackground And ObjectiveArtimisinin (an anti-malaria drug) is extracted from Artemisia annua and its water soluble derivative is dihydroartimisinin. Previous in vivo and in vitro studies showed that it has an inhibitory effect on T cells. It is also useful for immunosuppression of immune system.Materials And MethodsThe stable state kinetic for comparison of inhibitory effect of artimisinin, dihydroartimisinin and cyclosporine A on calcineurin activation was used in this study. First, the best inhibitory concentration of artimisinin, dihydroartimisinin and cyclosporine A was calculated. Afterwards, the km and Vmax of them in the presence of substrate, paranitrophenylphosphate (P-NPP) were measured. The KI was calculated in the presence of cyclosporine A, artemisinin and dihydroartemisinin. In this study, we used distilled water instead of sample in blank and cyclosporinA as a positive control group.ResultsThe Vmax in the control group was 81.97 M/min and in the presence of cyclosporine A and artemisinin or dihydroartemisinin were 81.97 M/min and 66.225 M/min, respectively. The Km in the absence of inhibitors was 1.886 M and in the presence of cyclosporine A and artemisinin or dihydroartemisinin was 2.819 M and 1.736, respectively. Also, KI in the presence of artimisinin and dihydroartimisinin was 4.219 ×10-5 M and in the presence of cyclosporine A was 2.021×10-5 M.ConclusionThis study indicates that the inhibitory power of artimisinin and dihydroartimisinin is almost equal and they inhibited calcineurin competitively, while inhibitory effect of cyclosporine A is non-competitive.Keywords: Cyclosporine A, Artimisinin, Dihydroartimisinin, Calcineurin, Calmudulin
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Background
Molecular deoxyribonucleic acid markers are one of the most important tools in molecular biology labs. The size of DNA molecule is determined by comparing them with known bands of markers during gel electrophoresis. In this study, we have suggested an efficient strategy to produce molecular weight markers in an industrial scale.
Materials and MethodsA combination of two previously known methods, restriction enzyme digestion and polymerase chain reaction (PCR), was used. The enzymatic digestion process was based on designing and constructing plasmids which equaled in size with the bands of ladder and produce the DNA fragment by plasmid linearization through digestion. In the PCR method, the DNA fragments with length 102 bp lesser than the related bands in DNA ladder are amplified by PCR and cloned in pTZ57T/A cloning vector. Then, PCRs with forward and reverse 100‑bp primers on the resulting plasmids amplify the ladder fragments. F100 and R100 primers bind to the backbone of pTZ57R (without insert) and amplify a 100‑bp PCR product. PCR on the plasmid with insert amplifies DNA fragment with 102+ insert length bp size.
ResultsUpon application of this strategy, 2000-10,000 bp DNA fragments were produced by enzymatic digestion of plasmids of the same size. Moreover, 100-1500 bp fragments were produced during PCR using only a set of forward and reverse (100 bp) primers.
ConclusionThe highest advantage of this cost–benefit approach is to produce different types of molecular weight markers by using an effective and short protocol.
Keywords: DNA ladder, DNA marker, molecular weight
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