Eman Mosad

Glomerular expression and elevated serum Bcl-2 and Fas proteins in lupus nephritis : preliminary findings

Programmed cell death (apoptosis) is involved in glomerular injuries leading to glomerulonephritis. Bcl‐2 and Fas are proteins that promote cell survival and death, respectively. This study tests the hypothesis that lupus nephritis is associated with alterations of Bcl‐2 and Fas protein expression. Thirty‐six patients with lupus nephritis and 10 controls (normal individuals) were included in this study. Bcl‐2 and Fas positive cells were examined in kidney biopsies by immunohistochemistry. Bcl‐2 and Fas serum levels were evaluated by enzyme‐linked immunosorbent assay (ELISA). In the glomeruli of normal kidneys, Bcl‐2 and Fas proteins were completely absent. In lupus nephritis patients, glomerular expression of Bcl‐2 and Fas was seen in mesangial cells (1·3 ± 0·1 and 2·0 ± 0·1 for Bcl‐2 and Fas, respectively). Similarly, a statistically significantly higher Bcl‐2 (217·1 ± 85·9) and Fas (767·9 ± 271) serum levels were found in lupus patients compared to controls (148·6 ± 87, 550·3 ± 91 for Bcl‐2 and Fas, P < 0·05). A direct correlation between serum Bcl‐2 and Fas and chronicity index was also found. Compared to normal controls, lupus nephritis is associated with glomerular expression and elevated serum levels of Bcl‐2 and Fas proteins. These findings suggest possible roles for Bcl‐2 and Fas in glomerular injury during evolution of lupus nephritis. The diagnostic, prognostic and therapeutic ramifications of our findings are open to further investigation.

Tissue Factor Pathway Inhibitor and P-Selectin as Markers of Sepsis-Induced Non-overt Disseminated Intravascular Coagulopathy

Inflammation and coagulation occur concomitantly in sepsis. Thrombin activates platelet that leads to P-selectin translocation, which upregulate tissue factor (TF) generation. Tissue factor pathway inhibitor (TFPI) is an anticoagulant that modulates coagulation induced by TF. The term non-overt disseminated intravascular coagulation (DIC) refers to a state of affairs prevalent before the occurrence of overt DIC. It was suggested that an initiation of treatment in non-overt DIC has better outcome than overt DIC. This study investigated the role of TFPI level, P-selectin, and thrombin activation markers in non-overt and overt DIC induced by sepsis and its relationship to outcome and organ dysfunction as measured by the Sequential Organ Failure Assessment (SOFA) score. It included 176 patients with sepsis. They were admitted to the pediatric intensive care unit (ICU).They included 144 cases of non-overt DIC and 32 cases of overt DIC. There was a significant difference in hemostatic markers, platelet count, partial thromboplastin time (PTT), P-selectin, thrombin activation markers, TFPI, and DIC score between overt and non-overt DIC in both groups. It was noticed that P-selectin was positively correlated with DIC score, fibrinogen consumption, fibrinolysis (D-dimer), thrombin activation markers, and TFPI. Tissue factor pathway inhibitor was significantly correlated with fibrinolysis, DIC score, and prothrombin fragment 1+2. Sequential Organ Failure Assessment score was correlated with DIC score and other hemostatic markers in patients with overt DIC. To improve the outcome of patients with DIC, there is a need to establish more diagnostic criteria for non-overt-DIC. Plasma levels of TFPI and P-selectin may be helpful in this respect.

Diagnostic performance and predictive value of rheumatoid factor, anti-cyclic-citrullinated peptide antibodies and HLA-DRB1 locus genes in rheumatoid arthritis

Background We evaluated the significance of the genes, defined as DRB1*04 or DRB1*01, in rheumatoid arthritis (RA) patients. We focused on the role of genetic and serologic markers to predict disease activity and destructive process of joints. Methods Sixty patients with RA were examined. Radiographic changes were evaluated by (Larsen score) and disease activity was measured by disease activity score 28 (DAS28). The markers analyzed were: erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), anti-cyclic citrullinated peptides (anti-CCP2) and HLA-DRB1 alleles typed by PCR. Results In this study, anti-CCP antibodies, CRP, RF and AKA were detected in 83.3%, 56.7%, 71.7% and 52% of patients respectively. HLA-DRB1*01 was found in 45% of patients and 35% of them had one or two HLA-DRB1*04 alleles. According to DRB1*04 subtypes, (DRB1* 0405) was present in of 80% them. For prediction of grade of activity, the independent predictors were anti-CCP (OR 19.6), and DRB1*04 positive allele (OR 5.1). The combination of DRB1*04 + anti-CCP antibodies gave increase in the specificity and positive predictive value to 92% and 90 respectively. As regards to the prediction of radiological joint damage, the independent predictors were HLA-DRB1*04, HLA-DRB1*01, RF, and CRP > 18 (OR 5.5, 4.5, 2.5, 2.0 respectively). Conclusion Our findings suggest that anti-CCP2 is superior to RF for the detection of RA and provided predictive information on joint destruction and disease activity. The presence of RA associated antibodies (ACCP or RF) and/or the SE genes are indicative for a poorer radiological outcome and higher grade of activity.

Bcl-XL and Bcl-2 expression in bilharzial squamous cell carcinoma of the urinary bladder: which protein is prognostic?

OBJECTIVES Bcl-2 and Bcl-XL are the most important antiapoptotic members of the Bcl-2 family frequently overexpressed in bladder cancer. Overexpression of Bcl-XL bilharzial-related bladder cancer was associated with tumor progression. However, the negative prognostic value of Bcl-2 expression is still questionable. This work studied the expression of Bcl-XL and Bcl-2 immunohistochemically in bilharzial-related squamous cell carcinoma of the urinary bladder and determined their prognostic value in relation to recurrence after radical cystectomy. METHODS A total of 72 patients with muscle-invasive bilharzial squamous cell carcinoma of the urinary bladder underwent radical cystectomy at our institution. The specimens were examined immunohistochemically for Bcl-XL and Bcl-2 expression. The patients were followed up for 3 years or until recurrence. The expression of Bcl-XL and Bcl-2 were related to the other prognostic indicators and patient survival. RESULTS The expressions of both Bcl-2 and Bcl-XL were significantly different according to the grade of malignancy. Bcl-XL expression was significantly related to tumor recurrence, but Bcl-2 expression was not. CONCLUSIONS To our knowledge, the present study is the first report of a negative prognostic value for Bcl-XL in bilharzial squamous cell carcinoma of the urinary bladder. However, this is another negative report on the prognostic value of bcl-2 in bilharzial bladder tumors.

Can Bcl-XL expression predict the radio sensitivity of Bilharzial-related squamous bladder carcinoma? a prospective comparative study

BackgroundLocal pelvic recurrence after radical cystectomy for muscle invasive bilharzial related squamous cell carcinoma accounts for 75% of treatment failures even in organ confined tumors. Despite the proven value of lymphadenectomy, up to 60% of patients undergoing cystectomy do not have it. These factors are in favor of adjuvant radiotherapy reevaluation. objectives: to evaluate the effect of adjuvant radiotherapy on disease free survival in muscle invasive bilharzial related squamous cell carcinoma of the urinary bladder and to test the predictability of radio-sensitivity using the anti apoptotic protein Bcl-XL.MethodsThe study prospectively included 71 patients, (47 males, 24 females) with muscle invasive bilharzial related squamous cell carcinoma of the bladder (Stage pT2a-T3N0-N3M0) who underwent radical cystectomy in Assiut university hospitals between January 2005 and December 2006. Thirty eight patients received adjuvant radiotherapy to the pelvis in the dose of 50Gy/25 fractions/5 weeks (Group 1), while 33 patients did not receive adjuvant radiotherapy (group 2). Immunohistochemical characterization for bcl-xL expression was done. Follow up was done every 3 months for 12 to 36 months with a mean of 16 ± 10 months. All data were analyzed using SPSS version 16. Three years cumulative disease free survival was calculated and adjusted to Bcl-XL expression and side effects of the treatment were recorded.ResultsThe disease free cumulative survival was 48% for group 1 and 29% for group 2 (log rank p value 0.03). The multivariate predictors of tumor recurrence were the positive Bcl-XL expression (odd ratio 41.1, 95% CI 8.4 - 102.3, p < 0.0001) and radiotherapy (odd ratio 0.19, 95% CI 0.05 - 0.78, p < 0.02). With Cox regression, the only independent multivariate predictor of radio-sensitivity was the Bcl-XL expression with odd ratio 4.6 and a p value < 0.0001. All patients tolerated the treatment with no life threatening or late complications during the period of follow up.ConclusionsAdjuvant radiotherapy for muscle invasive bilharzial related squamous cell carcinoma of the urinary bladder has potential effectiveness and minor side effects. Moreover, Bcl-XL expression is a valuable tool for predicting those who might not respond to this adjuvant treatment.

Persistence of TEL-AML1 fusion gene as minimal residual disease has no additive prognostic value in CD 10 positive B-acute lymphoblastic leukemia: a FISH study

ObjectivesWe have analyzed t(12;21)(p13:q22) in an attempt to evaluate the frequency and prognostic significance of TEL-AML1 fusion gene in patients with childhood CD 10 positive B-ALL by fluorescence in situ hybridization (FISH). Also, we have monitored the prognostic value of this gene as a minimal residual disease (MRD).MethodsAll bone marrow samples of eighty patients diagnosed as CD 10 positive B-ALL in South Egypt Cancer Institute were evaluated by fluorescence in situ hybridization (FISH) for t(12;21) in newly diagnosed cases and after morphological complete remission as a minimal residual disease (MRD). We determined the prognostic significance of TEL-AML1 fusion represented by disease course and survival.ResultsTEL-AML1 fusion gene was positive in (37.5%) in newly diagnosed patients. There was a significant correlation between TEL-AML1 fusion gene both at diagnosis (r = 0.5, P = 0.003) and as a MRD (r = 0.4, P = 0.01) with favorable course. Kaplan-Meier curve for the presence of TEL-AML1 fusion at the diagnosis was associated with a better probability of overall survival (OS); mean survival time was 47 ± 1 month, in contrast to 28 ± 5 month in its absence (P = 0.006). Also, the persistence at TEL-AML1 fusion as a MRD was not significantly associated with a better probability of OS; the mean survival time was 42 ± 2 months in the presence of MRD and it was 40 ± 1 months in its absence. So, persistence of TEL-AML1 fusion as a MRD had no additive prognostic value over its measurement at diagnosis in terms of predicting the probability of OS.ConclusionFor most patients, the presence of TEL-AML1 fusion gene at diagnosis suggests a favorable prognosis. The present study suggests that persistence of TEL-AML1 fusion as MRD has no additive prognostic value.

Changes in platelet, coagulation, and fibrinolytic activities in mitral stenosis after percutaneous mitral valvotomy: role of hemodynamic changes and systemic inflammation.

Markers of platelet activity (P-selectin), fibrinolysis (d-dimer), thrombin activity (prothrombin fragments 1, 2 [PF1,2] and thrombin–antithrombin III complex [TAT]), and inflammation (interleukin 1β [IL-1β]) were measured in 65 patients with mitral stenosis (MS) before and 2 weeks after percutaneous mitral valvotomy (PMV) and in 23 controls. All markers were significantly higher than the control and significantly decreased after PMV. P-selectin change correlated with the changes in left atrial diameter (LAD), pulmonary artery systolic pressure (PASP), and IL-1β. d-Dimer change had similar correlations, LAD, PASP, and IL-1β. The PF1,2 change correlated with the change in IL-1β. The TAT change correlated with the changes in LAD. The IL-1β change correlated with the changes in PASP. In conclusion, MS is associated with heightened inflammatory, platelet, thrombin, and fibrinolytic activities that decrease after PMV. Altered hemodynamics and reduced inflammatory activity might have a possible role in these changes.

Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P < 0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P < 0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

Fatigue in Rheumatoid Arthritis Patients: Association With Sleep Quality, Mood Status, and Disease Activity

OBJECTIVES Rheumatoid arthritis (RA) is a chronic inflammatory disease, characterized by polyarthritis and systemic manifestations. RA-fatigue is a significant problem and adds on disease burden. Sleep disturbance, depression, and disease activity are suggested contributing factors to RA-fatigue; however, their combined role did not examine before among Egyptian RA patients. The objective of the study was to investigate the presence of fatigue, sleep and mood disturbances in RA patients. Also, to evaluate the possible association of poor sleep, depression, and disease activity with RA-fatigue. METHODS This cross-sectional study included 115 RA patients diagnosed according to the 2010 ACR-EULAR criteria and 46 age and sex matched controls. Fatigue using the Multidimensional Assessment of Fatigue-Global Fatigue Index, sleep using the Pittsburgh Sleep Quality Index and mood status using Beck Depression Inventory were assessed for all participants. RA disease activity was evaluated using disease activity score-28 joints. RESULTS RA patients had higher mean fatigue, sleep disturbance, and depression scores (27.2±8.9, 6.4±3.6, and 12.8±7.3; respectively) than controls (22.7±7, 4.8±3, 7.8±5.9; respectively) (P<.05). Poor sleep, depression and higher disease activity were significantly correlated with fatigue (r=0.4, r=0.65, r=0.55; respectively) (P<.001). The three variables may explain up to 49.1% of the variation in fatigue on multiple regression analysis. CONCLUSION Fatigue, poor sleep, and depression are more common in Egyptian patients with RA. A remarkably higher fatigue was associated with poor sleep, depression, and high disease activity, thus monitoring these silent comorbidities in clinical practice is required.

FRI0665 Anti-heterogenous nuclear ribonucleoproteins (ANTI- HNRNP) and other autoantibodies for detection of erosive arthropathy in systemic lupus erythematosus with joint involvement in comparison by joint X-ray

Background Joint involvement in SLE is very common, affecting 90% of patients at some stage in the course of their disease (1).Arthritis featuring prominent radiological erosion in SLE is less common; however, in a small subset of patients an erosive pattern similar to RA develops (2). Anti- heterogenous nuclear ribonucleoprotein A2 (anti-hnRNP-A2) occur in about one-third of patients with RA but rarely in other arthritides such as OA, PsA or reactive arthritis. Interestingly, in SLE patients anti-hnRNP-A2 autoantibodies were found to be significantly associated with erosive arthritis (3). Objectives To investigate joint involvement in SLE and its relationship with autoantibodies to the hnRNP Ab A1 and A2, rheumatoid factor (R.F), Antinuclear antibody (A.N.A) and Anti-double stranded DNA (Anti-DSDNA) and correlation with articular involvement by joint x-ray. Methods Case series study comparing diagnosis of arthritis by hand and wrist x-ray with anti-hnRNP A1 and A2 in Fourty SLE patients aged 17–60 years old with disease duration 1–17 years complaining of arthralgia or arthritis. A controlled group of 21 clinically normal persons, age and sex matched and blood samples are drawn and centrifuged. RF and auto-antibodies to nuclear antigens anti-nuclear antibodies (ANA) and anti-double stranded DNA were determined in all the patients. All patients underwent X-rays of the hands and wrists. Results Anti-hnRNP A1 showed highly significant difference between study and control. Anti-hnRNP A2 showed significant difference between study and control.Table 1. Comparison of serological features in SLE patients with and without erosive arthritis (EA)* Characteristic SLE patients with EA SLE patients without EA Total P value Rheumatoid Factor, N (%)b 5 (50) 6 (20) 11 (27.5) 0.079 ANA, N (%)b 8 (80) 14 (46.7) 22 (55) 0.069 Anti-double stranded DNA, N (%)b 9 (90) 13 (43.3) 22 (55) 0.011*Table 2. Comparison of radiological findings of hand X-ray in SLE patients with and without erosive arthritis (EA)* Characteristic SLE patients with EA SLE patients without EA Total P value Juxta-articular osteoporosis, N (%) 9 (90) 21 (70) 30 (75) 0.204 Narrowing of joint space, N (%) 9 (90) 9 (30) 18 (45) 0.001* Subchondral cysts, N (%) 4 (40) 1 (3.3) 5 (12.5) 0.010* MCP sublaxation, N (%) 7 (70) 3 (10) 10 (25) 0.001* Interruption of cortical surface, N (%) 7 (70) 1 (3.3) 8 (20) <0.0001* New bone formation, N (%) 1 (10) 0 1 (2.5) 0.250 AVN, N (%) 0 1 (3.3) 1 (2.5) 0.750 Conclusions This study showed a high frequency of erosive arthropathy and autoantibody to both hnRNP antigens might become useful marker for joint involvement in SLE patients and identify SLE patients prone to develop joint damage References Pipili C, Sfritzeri A, Cholongitas E. Deforming arthropathy in systemic lupus erythematosus. Eur J Intern Med 2008;19(7):482–7. Pipili C, Sfritzeri A, Cholongitas E. Deforming arthropathy in SLE: review in the literature apropos of one case. Rheumatol Int 2009;29(10):1219–21. Hassfeld W, Steiner G, Graninger W, Witzmann G, Schweitzer H, Smolen JS. Autoantibody to the nuclear antigen RA33 – a marker for early rheumatoid arthritis.Br J Rheumatol 1993;86:19–203. Disclosure of Interest None declared