Assessment of circulating MCP-1 level and 2518A>G gene polymorphism in systemic lupus erythematosus

Lupus nephritis (LN) is a major contributor to morbidity and mortality in patients with Systemic Lupus Erythematosus (SLE). This study aims to investigate the possible role of a functional polymorphism in the regulatory region of the monocyte chemo-attractant protein-1 (MCP-1) gene and MCP-1 blood level in the diagnosis of LN and in correlating the MCP-1 blood levels with disease activity. The study included 56 SLE patients and 56 controls. All the SLE patients suffered from LN. An analysis of MCP-1 gene polymorphism by polymerase chain reaction was performed followed by restriction fragment length polymorphism (PCR-RFLP) analysis and MCP-1 blood level was determined using the ELISA technique. Calculation of Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) was performed. Serologic tests included the determination of antinuclear antibody (ANA) and anti-double-stranded (ds) DNA antibodies, Complement C3 and C4 levels. A significant increase in the frequency of genotype A/G and a decrease in the frequency of genotype A/A were found among patients with active LN compared to inactive LN. There was a statistically significant difference in the blood level of MCP-1 between LN patients and controls. Also, MCP-1 blood levels were significantly higher in active LN patients than inactive LN. A significant positive linear correlation was detected between MCP-1 blood level and SLEDAI, creatinine, and 24 hours protein in LN patients. These results suggest that an A/G genotype together with the measurement of the blood level of MCP-1 can be a useful tool for detection and follow up of active LN

Therapeutic potential of hydroxychloroquine on serum B-cell activating factor belonging to the tumor necrosis factor family (BAFF) in rheumatoid arthritis patients

Objective: To assess the serum B-cell activating factor belonging to the tumor necrosis factor family (BAFF) level in rheumatoid arthritis (RA) patients in view of different treatment regimens received and evaluate its relation with disease activity. Patients and methods: Ninety female RA patients were included. Sixty were on disease modifying anti-rheumatic drugs (DMARDs); 34 on hydroxychloroquine (HCQ) plus methotrexate (MTX), 26 on leflunomide (LFN) plus MTX and 30 newly diagnosed cases not yet on any treatment. Thirty age and gender matched healthy subjects served as controls. Full history taking, clinical examination and relevant laboratory investigations were performed. Disease activity score, in 28 joints (DAS-28), was calculated. Results: Serum BAFF level was significantly higher in patients (1.82 ± 0.91 ng/ml) compared to control (0.71 ± 0.33 ng/ml; p < 0.001). There was a significantly lower BAFF and disease activity in patients receiving DMARDs (1.55 ± 0.73 ng/ml and 3.08 ± 0.73) compared to new cases (2.36 ± 1.02 ng/ml and 3.46 ± 0.82) (p < 0.001 and p = 0.036, respectively). Those receiving HCQ + MTX had a lower BAFF level (1.29 ± 0.51 ng/ml) compared to those receiving LFN + MTX (1.94 ± 0.85 ng/ml; p = 0.002). The BAFF level significantly correlated with the presence of anti-CCP antibodies, DAS28 and MTX dose in all RA patients (r = 0.24, p = 0.02; r = 0.504, p < 0.001; r = 0.51, p < 0.001, respectively). Only DAS28 and MTX dose would highly influence the BAFF level (p = 0.015 and p = 0.001, respectively). Conclusion: Elevated level of BAFF in RA has been confirmed with a notable relation to disease activity making it a promising marker. The beneficial effect of hydroxychloroquine in dampening BAFF level throws light on the importance of considering it in combination among the newly developed biologics that also target B-cells

Relation of anti-annexin V antibodies to disease manifestations and activity in Behҫet’s disease patients

Aim of the work: To assess the role of serum anti-annexin V antibodies in Behҫet’s disease (BD) patients in relation to disease manifestations and activity. Patients and methods 65 BD patients and 30 matching controls were included. Disease activity was estimated by the Behçet Disease Current Activity Form (BDCAF). Serum IgG anti-annexin V antibodies titre was measured using the enzyme-linked immunosorbent assay. Results The patients’ age was 36.1 ± 8.5 years and disease duration 7.2 ± 5.2 years; 56 males and 9 females. The serum anti-annexin V antibodies level was significantly increased in the BD patients (50.9 ± 12.9 AU/ml) compared to the control (7.3 ± 3.1 AU/ml) (p < 0.0001). Serum anti-annexin V antibodies were significantly increased in BD patients with ocular involvement, skin lesions and neuro-Behcet’s compared to those without (p = 0.02, p = 0.004 and p = 0.002 respectively). Levels were comparable between those with uveitis, vitrous cells, retinal vasculitis, conjunctivitis and hypopyon and those without (p = 0.12, p = 0.22, p = 0.9, p = 0.67, p = 0.79 and p = 0.46 respectively). While those with xerophthalmia had a significantly higher level of anti-annexin V antibodies (60.6 ± 5.7 AU/ml) compared to those without (50.2 ± 13.1 AU/ml) (p = 0.02). The anti-annexin V antibodies significantly correlated with the BDCAF (r = 0.41, p = 0.001) and age (r = 0.43, p < 0.0001) but not with the disease duration (r = 0.22, p = 0.08), steroid dose (r = −0.21, p = 0.09) or laboratory investigations. On regression analysis, only the age would predict the anti-annexin V antibodies level (p = 0.02) while the BDCAF would not (p = 0.33). Conclusion There is a role of apoptosis in the pathogenesis of BD with special relation to the ocular, cutaneous and neurological manifestations and a possible link to the disease activity

Protective role of T regulatory (Treg) cells in systemic lupus erythematosus patients with nephritis

Aim: To assess the role of CD4+CD25+Foxp3+, CD4+CD25- Foxp3+, CD4+CD25+Foxp3- T cells and the fork head family transcription factor (Foxp3) gene polymorphism in systemic lupus erythematosus (SLE) and lupus nephritis (LN) patients. Patients and methods: The study comprised 40 SLE patients (including 38 females and 2 males) and 30 matched controls. SLE disease activity index (SLEDAI) was assessed. The percentage of expression of CD4, CD25 and Foxp3 on T cells was measured by flow cytometry. Foxp3 gene was genotyped using the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP). Results: The patients were 38 females and 2 males with a mean age of 30.3±8.9 years and disease duration of 7.4±5.7 years. Their mean SLEDAI was 7.5±6.7. Flow cytometry revealed 3 types of T cells; CD4+CD25+Foxp3+, CD4+CD25- Foxp3+ and CD4+CD25+Foxp3-. No significant differences were found in the percentage of expression and absolute counts of CD4+CD25+Foxp3+, CD4+CD25- Foxp3+ and CD4+CD25+Foxp3- T cells in patients and controls (p=0.26, p=0.22, p=0.32 respectively). CD4+CD25+Foxp3+T cells were significantly lower in patients with LN compared to those without (p<0.001). CD4+CD25- Foxp3+ T cells and Foxp3 (G/G) genotype were significantly associated with the grades of LN (p<0.001, p=0.003 respectively). The relationship between SLEDAI and the absolute counts of CD4+CD25+Foxp3+, CD4+CD25- Foxp3+ and CD4+CD25+Foxp3- T cells was insignificant (r=−0.04, p=0.78; r=0.05, p=0.76 and r=0.05, p=0.77 respectively). A significant difference in SLEDAI between patients expressing Foxp3 A/A genotype and those not was observed (p<0.01). Conclusion: CD4+CD25+Foxp3+, CD4+CD25- Foxp3+ and CD4+CD25+Foxp3- T cells are important regarding the pathogenesis of LN and could help assess the severity of renal involvement

Juvenile and juvenile-onset systemic lupus erythematosus patients: Clinical characteristics, disease activity and damage

Background: The diagnosis of systemic lupus erythematosus (SLE) in children is challenging. The heterogeneous manifestations and disease impact on the child’s growth highlight the importance of timely diagnosis and management. Objective: The aim of the work was to assess and compare the clinical characteristics, disease activity and damage between children with juvenile SLE (JSLE) and adult patients with juvenile-onset (JO-SLE). Patients and methods: 78 SLE patients; 26 children (JSLE) and 52 JO-SLE adults were included in this study. Disease activity was assessed using the SLE Disease Activity Index (SLEDAI) and organ damage using the Systemic Lupus International Collaborating Clinics (SLICC) index. Results: The mean age of the JSLE children was 13.25 ± 2.09 years and 23.17 ± 4.26 years for JO-SLE cases. Age at disease-onset and female gender tended to be higher in JO-SLE cases than in children with JSLE. There was a significantly higher frequency of serositis, nephritis and hematological involvement in the JO-SLE (57.7%, 76.9%, 73.1%, respectively) compared to the JSLE cases (15.4%, 30.8%, 30.8%, respectively) (p < 0.001 for all). The erythrocyte sedimentation rate, creatinine and proteinuria were significantly increased in JO-SLE while alkaline phosphatase was higher in JSLE cases. In JO-SLE cases, SLEDAI significantly increased (5.96 ± 6.18 vs 3.12 ± 1.97; p = 0.003) and the SLICC tended to increase compared to the JSLE children. More JO-SLE cases received hydroxychloroquine and azathioprine. Conclusion: The existence of differences in clinical phenotype has been confirmed, between JSLE and JO-SLE especially as regards serositis, nephritis and heamatological affection. The disease damage was comparable which denotes that the maximum organ involvement occurs in childhood with an almost stationary course

Identification of Antibacterial Metabolites from Endophytic Fungus Aspergillus fumigatus, Isolated from Albizia lucidior Leaves (Fabaceae), Utilizing Metabolomic and Molecular Docking Techniques

The rapid spread of bacterial infection caused by Staphylococcus aureus has become a problem to public health despite the presence of past trials devoted to controlling the infection. Thus, the current study aimed to explore the chemical composition of the extract of endophytic fungus Aspergillus fumigatus, isolated from Albizia lucidior leaves, and investigate the antimicrobial activity of isolated metabolites and their probable mode of actions. The chemical investigation of the fungal extract via UPLC/MS/MS led to the identification of at least forty-two metabolites, as well as the isolation and complete characterization of eight reported metabolites. The antibacterial activities of isolated metabolites were assessed against S. aureus using agar disc diffusion and microplate dilution methods. Compounds ergosterol, helvolic acid and monomethyl sulochrin-4-sulphate showed minimal inhibitory concentration (MIC) values of 15.63, 1.95 and 3.90 &micro;g/mL, respectively, compared to ciprofloxacin. We also report the inhibitory activity of the fungal extract on DNA gyrase and topoisomerase IV, which led us to perform molecular docking using the three most active compounds isolated from the extract against both enzymes. These active compounds had the required structural features for S. aureus DNA gyrase and topoisomerase IV inhibition, evidenced via molecular docking

Anticholinesterase Activity of Budmunchiamine Alkaloids Revealed by Comparative Chemical Profiling of Two <i>Albizia</i> spp., Molecular Docking and Dynamic Studies

Alzheimer’s disease remains a global health challenge and an unmet need requiring innovative approaches to discover new drugs. The current study aimed to investigate the inhibitory activity of Albizia lucidior and Albizia procera leaves against acetylcholinesterase enzyme in vitro and explore their chemical compositions. Metabolic profiling of the bioactive plant, A. lucidior, via UHPLC/MS/MS-based Molecular Networking highlighted the richness of its ethanolic extract with budmunchiamine alkaloids, fourteen budmunchiamine alkaloids as well as four new putative ones were tentatively identified for the first time in A. lucidior. Pursuing these alkaloids in the fractions of A. lucidior extract via molecular networking revealed that alkaloids were mainly concentrated in the ethyl acetate fraction. In agreement, the alkaloid-rich fraction showed the most promising anticholinesterase activity (IC50 5.26 µg/mL) versus the ethanolic extract and ethyl acetate fraction of A. lucidior (IC50 24.89 and 6.90 µg/mL, respectively), compared to donepezil (IC50 3.90 µg/mL). Furthermore, deep in silico studies of tentatively identified alkaloids of A. lucidior were performed. Notably, normethyl budmunchiamine K revealed superior stability and receptor binding affinity compared to the two used references: donepezil and the co-crystallized inhibitor (MF2 700). This was concluded based on molecular docking, molecular dynamics simulations and molecular mechanics generalized born/solvent accessibility (MM–GBSA) calculations