Circulating Levels of Vascular Endothelial Growth Factor and Basic Fibroblastic Growth Factor in Infantile Hemangioma Versus Vascular Malformations.

Few preliminary reports studied the utility of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) for differentiation between infantile hemangiomas (IHs) and vascular malformations. The aim of this study was to investigate the role of serum VEGF and bFGF levels in differentiating IHs from vascular malformations and identifying the stage and clinical course of IHs. Serum levels of VEGF and bFGF were assessed in 60 infants and children with various cutaneous vascular anomalies defined in 3 groups: proliferating IHs (n = 25), involuting IHs (n = 23), and vascular malformations (n = 12), in comparison with their levels in 40 healthy matched control. Serum levels of VEGF and bFGF were significantly elevated in all groups as compared to control ( P < .001, respectively). Both proliferating and involuting IHs had comparable levels of both markers ( P > .05, respectively) that were significantly higher in comparison with vascular malformations ( P < .05, respectively). Significantly lower VEGF levels were found in IHs that had regressed spontaneously (n = 11) compared to those regressed by treatment (n = 37), ( P < .05); meanwhile, bFGF showed no significant difference between both groups ( P > .05). Using receiver operating characteristic curves, a combined use of VGEF and bFGF yielded a sensitivity of 85.42% and a specificity of 100% for differentiating IHs from vascular malformations. Serum VEGF and/or bFGF levels are increased in cutaneous vascular anomalies and can differentiate IHs from vascular malformations. None of these markers could help in identifying the stage of IHs. Low VEGF is associated with spontaneous regression of IHs.

Circulating CD4(+) CD25(high) FoxP3(+) T-regulatory cells in patients with atopic dermatitis after narrowband-ultraviolet B phototherapy.

BACKGROUND: Previous studies showed controversial results regarding CD4(+) CD25(high) FoxP3(+) T-regulatory cells (Tregs) in atopic dermatitis (AD) and effect of therapy. METHODS: Circulating CD4(+) CD25(high) FoxP3(+) Tregs were assessed by flow cytometry in 20 controls and 20 patients with AD at baseline and after narrowband ultraviolet B with assessment of disease severity. RESULTS: Patients showed higher pretreatment T-effector cells (Teffs) (%) and lower pretreatment Tregs FoxP3 expression% than controls (P = 0.003 and 0.01, respectively). Mild AD showed a lower Tregs/Teffs ratio compared to controls (P = 0.013), while moderate group showed higher Teffs%, and lower Tregs FoxP3 expression% and Tregs/Teffs compared to controls (P = 0.016, 0.007, and 0.009 respectively). The severe group had higher Tregs% and Teffs%, yet with a lower Tregs FoxP3 expression% compared to controls (P < 0.001, P = 0.043, P = 0.044, respectively). There was significant reduction of severity after narrowband ultraviolet B (P = 0.007), with overall significant elevation of Tregs FoxP3 expression% in patients (P = 0.004). All patients' post-treatment laboratory findings were statistically matched to each other and to controls whatever their previous severity or therapeutic response. The improvement of severity score correlated with the change in both Tregs% and Tregs/Teffs. CONCLUSIONS: Significant reduction in AD disease severity is correlated with the change in Tregs% and Tregs/Teffs.

Detection of 14q32 rearrangements in multiple myeloma, using simultaneous FISH analysis combined with immunofluorescence.

BACKGROUND: 14q32 rearrangement has been identified as a recurrent hotspot of translocations in multiple myeloma (MM). The Fluorescence Immunophenotyping and Interphase Cytogenetics as a tool for the Investigation of Neoplasms (known as FICTION technique) for evaluation of chromosomal changes in MM. The aim of this work is to detect 14q32 rearrangement, using FICTION technique, on archival bone marrow (BM) slides of MM patients, and to study its prognostic value. METHOD: This study was conducted at Ain Shams University Hospital. The FICTION technique, which uses CD138 and dual color, and the break-apart 14q32 rearrangement probe, was performed on archived smears of BM slides for 50 MM patients at the time of diagnosis. RESULTS: A significantly higher percentage of cases were positive for 14q32 rearrangement by FICTION (32%) compared to fluorescence in situ hybridization (FISH) (12%) (p=0.04). Cases positive by FICTION for the rearrangement were designated as Group A, while negative cases were designated as Group B. Significantly lower Hb and CRP levels were found among Group B when compared to Group A patients (p=0.001 and 0.01, respectively). Serum albumin level and Bence Jones protein (BJP) significantly affect overall survival (OS) (p=0.01, 0.007, respectively). However, a statistically non-significant shorter mean survival time was found in positive cases through FICTION versus negative cases. CONCLUSION: FICTION technique provides a sensitive tool for establishing clonal plasma cells (PC) infiltration of BM aspirates, and is amenable for use on archived as well as fresh smears.

Impact of CYP2C9 and VKORC1 genetic polymorphisms upon warfarin dose requirements in Egyptian patients with acute coronary syndrome.

Warfarin is the most widely prescribed anticoagulant drugs. Cytochrome P450-2C9 (CYP2C9) and vitamin K epoxide reductase-oxidaxe complex subunit 1 (VKORC1) contribute significantly to the variability of warfarin dose requirements among patients. We investigated the impact of CYP2C9 and VKORC1 polymorphisms on the variability of warfarin dosage requirements in Egyptian patients with acute coronary syndrome and their association with other nongenetic factors. Eighty participants with acute coronary syndrome were enrolled in this cross-sectional study. Associations between CYP2C9 and VKORC1 gene variants together with daily warfarin dose, demographic data, clinical status of patients and time to target international normalized ratio were assessed. Mean warfarin dose among patients with wild-type CYP2C91/1 genotype was significantly higher than heterozygous CYP2C91/2 and CYP2C91/3 variants (P ≤ 0.001). Patients with wild VKORC1 (G/G) genotype were treated with significantly higher daily warfarin dosages than homozygous (A/A) and heterozygous (G/A) genotypes. Patients carrying VKORC1 (G/G) genotype in combination with the CYP2C91/1 type alleles had the highest daily warfarin dosage, whereas the lowest daily warfarin dosage to achieve the required clinical effect was found among patients having CYP2C91/2 and CYP2C91/3 genotypes combined with VKORC1 (A/A) genotype (P ≤ 0.001). Regression analysis revealed that age, height, CYP2C9 and VKORC1 genotypes were significantly associated with warfarin dose. Genetic polymorphisms in VKORC1, CYP2C9 along with age and height are determinants of warfarin dose requirements in Egyptian population acute coronary syndrome. Higher warfarin loading dose is required for both wild CYP2C9 and VKORC1 gene variants which may contribute to warfarin-resistant cases.

Alterations of platelet functions in children and adolescents with iron-deficiency anemia and response to therapy.

Several changes in platelets have been reported in patients with iron-deficiency anemia (IDA), so a relationship between iron metabolism and thrombopoiesis should be considered. We aimed to study the alterations of platelet functions in patients with IDA by assessment of platelet aggregation with epinephrine, adenosine diphosphate (ADP) and ristocetin and by measuring platelet function analyzer-100 (PFA-100) closure time together with the effect of iron therapy on the same tests. A follow-up study was conducted in Ain Shams University Children's hospital in the period from June 2011 to June 2012 including 20 patients with confirmed IDA and 20 healthy age- and sex-matched control. Bleeding manifestations were reported. Laboratory analysis included complete blood count, assessment of iron status by measuring serum iron, TIBC and ferritin, assessment of platelet functions by PFA-100 closure time and platelet aggregation with collagen, ADP and ristocetin. Patients with IDA were treated by oral iron therapy 6 mg/kg/day of ferrous sulfate and post-therapeutic re-assessment was done. Mean age of IDA patients was 5.7 ± 4.2 years. Bleeding manifestations were more common in patients group. Mean PFA-100 closure times (with epinephrine) were significantly longer in patients (179.1 ± 86.4 seconds) compared to control group (115 ± 28.5 seconds) (p < 0.05). Platelet aggregation by ADP (38.1 ± 22.2%), epinephrine (19.7 ± 14.2%) and ristocetin (58.8 ± 21.4%) were significantly reduced in patients compared to control (62.7 ± 6.2, 63.3 ± 6.9, 73.8 ± 8.3, respectively; p < 0.001). After treatment platelet aggregation tests induced by ADP (64.78 ± 18.25%), and epinephrine (55.47 ± 24%) were significantly increased in patients with IDA compared to before treatment (39.44 ± 21.85%, 20.33 ± 14.58%; p < 0.001). PFA-100 closure time as well showed significant decreased after treatment (118.4 ± 27.242) compared to before treatment (186.2 ± 90.35; p < 0.05). A negative correlation between platelet aggregation induced by ADP and mean values of serum ferritin before treatment (r = 0.042, p < 0.05) was found. A mutual effect is considered between iron deficiency and platelet functions. Subtle bleeding manifestations can occur in patients with IDA with delay in platelet aggregation and prolongation in PFA-100 closure times which can be reversed by iron therapy.

Serum Th1/Th2 and macrophage lineage cytokines in leprosy; correlation with circulating CD4(+) CD25(high) FoxP3(+) T-regs cells.

Not only macrophages, T-helper (Th)1 and Th2, but also CD4(+) CD25(high) FoxP3(+) regulatory T cells (T-regs) are involved in immune response to Mycobacterium leprae. We aimed to evaluate serum interleukin (IL)-1ß and IL-12p70 (macrophage cytokines), interferon-γ (IFN-γ) (Th1 cytokine), IL-4 (Th2 cytokine) and circulating CD4(+) CD25(high) FoxP3(+) T-regs, in untreated leprosy patients. Forty three patients and 40 controls were assessed for the mentioned cytokines using ELISA. Patients were assessed for circulating T-regs using flow cytometry. Patients were subgrouped into tuberculoid (TT), pure neural leprosy (PNL), borderline cases, lepromatous (LL), type 1 reactional leprosy (RL1) and erythema nodosum leprosum (ENL). Serum IL-12p70, IFN-γ and IL-4 were significantly higher in patients versus controls (P < 0.05). Serum IL-4 was highest in LL and lowest in RL1 (P = 0.003). Serum IL-1ß levels was significantly higher in multibacillary versus paucibacillary patients (P = 0.006). Significantly higher T-regs levels was detected in TT, RL1 and PNL, while the lowest levels in ENL(P < 0.001), with significant differences versus controls (P < 0.05). FoxP3 expression% was significantly lower in PNL than other patients and controls (P < 0.05). T-regs/T-effs was lowest in ENL(P < 0.05). IFN-γ correlated positively with T-regs but negatively with IL-1ß (P = 0.041&0.046 respectively), which correlated positively with T-effs%( P = 0.05). IL-4 correlated positively with T-regs FoxP3 expression% (P = 0.009). We concluded that: Circulating T-regs were increased in TT, RL1 and PNL patients, known of relatively high cell-mediated immunity. This finding was supported by low FoxP3 expression (in PNL) and correlation between T-regs count and IFN-γ level. Overproduction of IL-4 in LL may infer liability to develop ENL, with disease progression and immune hyperactivation, marked by deficient T-regs and increased T-regs FoxP3 expression%. IL-1ß probably has a pro-inflammatory role in multibacillary patients as correlated with T-effs%.

The degree of safety of family replacement donors versus voluntary non-remunerated donors in an Egyptian population: a comparative study.

BACKGROUND: Screening donated blood for transfusion-transmissible infections is considered an important strategy for maximising the safety of blood transfusions. MATERIALS AND METHODS: A total of 17,118 donors, classified into two groups--family replacement donors and voluntary non-remunerated donors--were investigated for hepatitis B virus (HBV) surface antigen and antibodies against hepatitis C virus (HCV), human immunodeficiency virus (HIV) and Treponema pallidum. In addition cytomegalovirus (CMV) antibodies were searched for in 160 donors (80 from each group). All the laboratory tests were done using enzyme-linked immunosorbent assays. RESULTS: Of the total cohort of donors, 87.7% were family donors and 12.3% were voluntary non-remunerated donors. There was a highly significant difference in age and gender between the two types of donors with voluntary donors being much younger and including a much higher proportion of male donors than female donors. The prevalences of HBV, HCV and CMV IgG were much higher in family donors than in voluntary donors, with the differences being highly statistically significant. There was also a significantly higher prevalence of syphilis among family replacement donors. As regards HIV and CMV IgM, significant differences were not detected between the two groups. DISCUSSION: The results of our study clearly showed that the prevalence of transfusion-transmissible infections is much higher among family replacement donors than among voluntary donors, and that voluntary donors are the best way of achieving safer blood.

Antenatal administration of vitamin K1: relationship to vitamin K-dependent coagulation factors and incidence rate of periventricular-intraventricular hemorrhage in preterm infants; Egyptian randomized controlled trial.

OBJECTIVES: To determine the effect of maternal antenatal administration of vitamin K1 on activity level of vitamin K-dependent coagulation factors and on the occurrence of periventricular-intraventricular hemorrhage (PIVH). METHODS: This study was conducted on 90 infants who were classified into; Group A: 30 preterm whose mothers received antenatal vitamin K1, Group B: 30 preterm whose mothers did not receive antenatal vitamin K1, and Group C: 30 healthy full term newborns as a control group. All newborns were subjected to measurement of the activity level of vitamin K-dependent coagulation factors (FII, FVII, FIX and FX). Cranial ultrasound was done on the 1st, 3rd and 7th days of life. RESULTS: Group B showed significantly lower activity level of FII and FX with higher incidence of PIVH compared with group A. Neonates who developed PIVH by the 7th day in both group A and B had significantly lower activity level of vitamin K-dependent coagulation factors. CONCLUSION: when antenatal vitamin K1 was given to pregnant women at imminent risk of preterm labor, their preterm neonates were able to achieve a clotting status approaching that of full term neonates and are less liable to develop PIVH.

Clinical significance of baseline platelet reactivity in patients with acute coronary syndrome.

UNLABELLED: The aim of this study was to assess the baseline platelet hyperactivity in patients with acute coronary syndrome (ACS) and correlate it with the disease severity and clinical outcome. PARTICIPANTS AND METHODS: This study was conducted on 60 patients with the first attack of non-ST-segment elevation ACS and 30 healthy controls. Patients were subdivided into 2 groups: group I: 40 patients with non-ST-segment elevation myocardial infarction (NSTEMI) and group II: 20 patients with unstable angina. All the studied patients were subjected to baseline platelet function analysis using platelet function analyzer 100 (PFA-100). RESULTS: Group I patients showed significantly lower PFA-100 closure time (CT) than group II (P = .04) and control group (P = .03). The occurrence of complications in NSTEMI patients was associated with older age, hypertension, and shorter baseline collagen adenosine diphosphate PFA-100 CT (P < .05). CONCLUSION: NSTEMI patients proved to have enhanced platelet function that can be used as a predictor of the outcome.

In vivo effect of recombinant human granulocyte colony-stimulating factor on neutrophilic expression of CD11b in septic neonates: a randomized controlled trial.

Neonates are susceptible to septicemia secondary to quantitative and qualitative neutrophilic defects. Granulocyte colony-stimulating factor (G-CSF) stimulates myeloid progenitor cell proliferation and induces selective neutrophil functions. The authors aimed to evaluate the effect of G-CSF administration in septic neonates on neutrophil production and CD11b expression. Sixty septic neonates were randomized to receive intravenous G-CSF 10 µg/kg/day for 3 days (G-CSF group, n = 30), or not to receive G-CSF (non-G-CSF group, n = 30). Thirty healthy newborns were included as controls. Laboratory investigations included complete blood count, C-reactive protein, blood culture, renal and liver function tests, and assessment of neutrophilic expression of CD11b. Total leukocytes count (TLC), absolute neutrophil count (ANC), and immature myeloid cell count in G-CSF group showed significant difference between post-and pre-G-CSF levels. TLC, ANC, immature myeloid cell count and immature/total myeloid cells ratio were higher in G-CSF group compared to non-G-CSF group on days 1 and 3. Higher neutrophilic expression of CD11b was reported in both septic groups on day 0 compared to control group. On day 5, CD11b was higher in G-CSF group than non-G-CSF group. G-CSF improved CD11b% in neutropenic and non-neutropenic septic neonates. No significant difference was found between pre- and posttreatment renal and liver function tests. Lower duration of antibiotic intake and hospitalization was observed in G-CSF group compared to non-G-CSF group. G-CSF administration as an adjuvant therapy for neonatal septicemia, whether neutropenic or not, improves neutrophilic count and function and contributed to early healing from sepsis.

Assessment of osteoporosis in psoriasis with and without arthritis: correlation with disease severity.

BACKGROUND: The most frequent extracutaneous association with psoriasis is arthritis. Because proinflammatory cytokines are increased in psoriasis, patients with this disease may be more prone to osteoporosis than the healthy individuals. METHODS: We evaluated 50 patients with psoriasis, with or without psoriatic arthritis (PsA), for the presence and degree of osteoporosis by performing dual energy x-ray absorptiometry (DEXA) and obtaining serum osteoprotegrin (OPG) levels. In addition, we correlated these results with the extent of skin and joint disease. Psoriasis area and severity index (PASI) was determined in all 50 patients with psoriasis, and total joint score (TJS) was recorded in the 16 patients who also had PsA. Results of DEXA and serum OPG were also obtained for 20 healthy individuals who served as controls. RESULTS: Osteoprotegrin level was significantly increased in psoriasis patients (with or without PsA) vs. controls. However, DEXA revealed that PsA patients had a higher degree of osteoporosis in the femur neck and wrist. In PsA patients, TJS correlated positively with both disease duration and PASI but correlated negatively with Z score of the femur. CONCLUSION: Psoriasis patients with or without arthritis may suffer from osteoporosis as evidenced by significantly increased serum OPG. Prolonged and extensive cutaneous disease is an important risk factor for the development and severity of PsA. Patients with a greater number of affected joints are at higher risk of osteoporosis.

Circulating CD4+ CD25 high FoxP3+ T cells vary in different clinical forms of leprosy.

BACKGROUND: CD4(+) CD25(high) FoxP3(+) regulatory T cells (T-regs) were reported to increase in chronic infections. We aimed at studying their frequency in leprosy to investigate their role during Mycobacterium leprae infection. METHODS: Using flow cytometry, the frequency and FoxP3 expression of circulating T-regs was assessed in 38 leprosy patients and 38 healthy controls. Patients were divided into; group I tuberculoid (TT), group II borderline cases [borderline tuberculoid (BT), borderline (BB), and borderline lepromatous (BL)], group III lepromatous (LL), and group IV erythema nodosum leprosum (ENL). RESULTS: Mean T-regs% and FoxP3 expression were significantly elevated in patients (particularly TT) compared to controls (3.8 ± 2.5% vs. 2.5 ± 0.8% and 78.8 ± 56.2% vs. 55.8 ± 15.7%, respectively) (P < 0.05). Comparing the four disease groups, T-regs% was significantly different (median 5.3% in group I, 3.4% in group II, 2.8% in group III, and 1.2% in group IV; P = 0.005). FoxP3% on T-regs was not significantly different between them [median 71.5% in TT, 62.3% in borderline categories, 67.75% in LL, and 85.75% in ENL; P = 0.149). Notably FoxP3 expression was significantly higher in ENL than controls (P = 0.011). CONCLUSION: The frequency and suppressive marker of circulating T-regs are elevated in TT patients. Patients with LL and ENL express significantly lower frequency of T-regs and higher FoxP3 expression (in ENL), consistent with disease progression and immune hyper-activation in these disease categories. Thus, rather than being detrimental to immunity, intact T-regs activity may be beneficial to leprosy patients.

Prognostic significance of E-cadherin expression and peripheral blood micrometastasis in gastric carcinoma patients.

BACKGROUND: The primary objective is to evaluate the prognostic value of E-cadherin (E-cad) expression and peripheral blood micrometastasis (PBMM) in gastric carcinoma. Secondary objective is to study the association between these 2 markers and the clinicopathological features of the patients. MATERIALS AND METHODS: This study took place at Ain Shams University Hospitals. A total of 30 patients with histologically proven gastric adenocarcinoma after curative surgical resection were enrolled in this study. E-cad expression was assessed in tumor tissue samples. Before the start of adjuvant chemoradiotherapy, fresh blood samples were collected to detect PBMM as indicated by cytokeratin18 mRNA expression using real-time quantitative polymerase chain reaction (RQ-PCR). RESULTS: Both abnormal E-cad expression and PBMM were significantly associated with lymph node metastasis, TNM stage, and lymphatic invasion. Moreover, PBMM was significantly associated with poor tissue differentiation and vascular invasion (P < .05). We found strong agreement between E-cad expression and presence of PBMM (P = .001). Both cases with altered E-cad expression and cases with positive PPMM showed shorter relapse-free survival (RFS) (P = .003 and <.001, respectively). Cox regression analysis showed that positive PBMM was independent predictor factor for relapse (hazard ratio [HR] = 6.14; 95% confidence interval [95% CI] = 1.06-35.63; P = .04). Cases with positive PBMM showed shorter overall survival (OS) (P = .001). CONCLUSIONS: In conclusion, loss of normal E-cad expression in gastric cancer showed a close correlation with the presence of PBMM. PBMM was associated with poor RFS independent of other clinicopathological features. Additionally, detection of PBMM was a significant indicator of OS, and intensive chemotherapy seems to be indicated for these patients.

Prognostic value of immunohistochemical classification of diffuse large B-cell lymphoma into germinal center B-cell and non-germinal center B-cell subtypes.

OBJECTIVE: To study the expression of germinal center B-cell (GCB)/activated B-cell like-related proteins to get optimal stratification of diffuse large B-cell lymphoma (DLBCL) patients, and correlate this with the established clinical and laboratory parameters. METHODS: This study was conducted retrospectively on 30 archival paraffin tissue blocks of DLBCL. All patients were diagnosed between April 2004 and January 2007 at Ain Shams University Hospital and National Cancer Institute, Cairo, Egypt. All patients received anthracycline-based regimens, and none of them received rituximab immunotherapy. Each case included in this study was investigated by immunohistochemical reaction for multiple myeloma-1/interferon regulatory factor-4, B-cell/lymphoma 6, and cluster of differentiation10 monoclonal antibodies. RESULTS: Patients were classified as GCB group (17 patients) and non-GCB group (13 patients). We found a statistically significant association between non-GCB phenotype and performance status (PS) more than 1, high lactate dehydrogenase (LDH) level, advanced international prognostic index (IPI), and poor patient outcome. Non-GCB phenotype, high LDH level, and PS more than 1 were all associated with increased mortality risk. The median survival time was 46.9 months in group A compared to 19.6 months in group B (hazard ratio[HR]=3.30; 95% confidence interval [CI]=0.52-21.10). Using multivariate Cox regression analysis, non-GCB phenotype was found to be the most predicting factor (HR=6.07; 95% CI=1.6-22.9; p=0.008). CONCLUSION: The subclassification of DLBCL into GCB and non-GCB groups using immunohistochemistry may be useful for identifying those patients whose prognosis is so poor that more aggressive therapy can be given at the time of diagnosis.

Tissue Factor, Tissue Factor Pathway Inhibitor and Factor VII Activity in Cardiovascular Complicated Type 2 Diabetes Mellitus.

OBJECTIVES: Tissue factor (TF) is the main initiator of the extrinsic coagulation pathway through factor VII (FVII) activation, which is physiologically inhibited by tissue factor pathway inhibitor (TFPI). Alteration of this pathway has been described in Type 2 diabetes mellitus (T2DM). The aim of this study is to assess TF and TFPI plasma levels and FVII coagulant activity (FVIIa) in T2DM in relation to cardiothrombotic disease and their correlation to metabolic and clinical behavior of the patients. METHODS: The study was conducted on 80 T2DM patients divided to accordingly; groupI: 40 patients without a history or clinically detected heart disease, and groupII: 40 patients with a history of myocardial infarction compared to 30 controls. The patients were recruited from Ain Shams University diabetes clinic from September 2007 to February 2009 after informed consent was obtained. Peripheral blood samples were taken for measurement of plasma TF and TFPI levels using ELISA technique and quantitative FVIIa using FVII deficient plasma. RESULTS: Plasma levels of TF, TFPI and FVIIa were significantly higher in T2DM patients compared to the controls (p<0.001). TF (236.50±79.23)and TFPI (242.33±85.84)were significantly higher in group II, compared to group I (150.33±81.16), (152.8± 82.46), (p<0.001). TF and TFPI were significantly correlated to body mass index and glycemic control. Also, TF and TFPI were significantly higher in hypertensives (p=0.001) and dyslipidemics (p=0.006) but not in smokers (p=0.64), (p=0.11) respectively. CONCLUSION: There was a correlation between high TF, TFPI plasma levels, FVIIa activity and cardiothrombotic complications in T2DM especially in the presence of high risk factors such as poor glycemic control, dyslipidemia and obesity. Future target therapy against TF may be beneficial for T2DM patients.