Wael Alkhiary

Association of Hemostatic Gene Polymorphisms With Early-Onset Ischemic Heart Disease in Egyptian Patients

The association between hereditary thrombophilia and venous thrombosis is well established but controversial data exist with respect to arterial thrombosis. We performed a pilot study on 31 patients with acute myocardial infarction (AMI), 21 patients with unstable angina (UA), and 20 healthy volunteers to investigate the role of various hemostatic gene polymorphisms in young Egyptian patients, who survived their first ischemic heart disease (IHD). Thrombophilic gene polymorphisms were tested using multiplex polymerase chain reaction and reverse-hybridization technique. We showed an increased risk of AMI with factor V (FV) Leiden and prothrombin G20210A heterozygosity. The increased risks of UA was associated with GA and A allele of fibrinogen β-455G→A polymorphism. Conversely, factor XIII (FXIII) Val34Leu GT and T allele were protective in the UA group. Nevertheless, the prevalence of FV H1299R, plasminogen activator inhibitor 1 4G/5G, glycoprotein IIIa C1565T, 5,10-methylenetetrahydrofolate reductase C677T, and A1298C mutations did not differ between patients with IHD and controls. The data have clinical implications regarding screening and thromboprophylaxis in high-risk individuals younger than 40 years.

Antiplatelet and Anticoagulant Activities of Angelica shikokiana Extract and Its Isolated Compounds

Angelica shikokiana is a Japanese medicinal plant that is used traditionally in several ailments of cardiovascular diseases. However, there is no report regarding its anticoagulant or antiplatelet activities. So this study was designed to screen for such activities (anticoagulant by prothrombin time [PT], activated partial thromboplastin time, and thrombin time assays and antiplatelet activities against adenosine 5′-diphosphate [ADP] and arachidonic acid-induced platelet aggregations) for the methanol extract of the aerial part (Angelica methanol extract [AME]), its isolated coumarins, flavonoids, and flavonoid metabolites. The AME had potent anticoagulant and antiplatelet activities, and the flavonoid compounds were evidenced to be responsible for such activities. Among coumarins compounds, hyuganin C showed significant prolongation of only PT, while other coumarins were inactive. Similarly, hyuganin C and bergapten were the only active coumarins against ADP-induced platelet aggregation. Compared to the parent compounds, colonic metabolites of the flavonoids had similar anticoagulant and antiplatelet activities, while glucuronides showed sharp decreases in all studied activities. This is the first report showing that the medicinal plant A shikokiana has potent antiplatelet and anticoagulant activities.

Tumor necrosis factor-α (TNF-α) −308 G/A and lymphotoxin-α (LT-α) +252 A/G genetic polymorphisms in Egyptian acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is one of the most hematological malignancies of lymphoid origin. It has been proposed that deregulation of cytokines could be linked with pathogenesis, progression, and survival in many diseases. Genetic polymorphisms in two important cytokines like tumor necrosis factor-α (TNF-α) −308 and lymphotoxin-α (LT-α) +252 can disturb both their transcription and expression and lead to their high plasma levels. A difference in the occurrence of the polymorphisms in TNF-α −308 G/A and LT-α +252 A/G in ALL cases among several populations with different ethnicities was observed. The study investigated the occurrence and the role of polymorphisms of tumor necrosis factor genes including TNF-α −308 G>A and LT-α +252 A>G in the development of ALL in Egypt. A case-control study was done on 126 newly diagnosed ALL patients (96 pediatric and 30 adult patients); 130 healthy subjects composed the control group. Polymorphism variants of TNF-α and LT-α genes were studied by PCR-RFLP on genomic DNA of all studied individuals. TNF-α −308 G/A polymorphism was statistically significant in ALL pediatric patients (P value = 0.008) with no association with ALL adult patients. TNF AA homozygous variant genotype and the A allele both showed significant risks of the development of pediatric ALL. However, there was no association between LT-α +252 A/G polymorphism in both pediatric and adult ALL. The results show that TNF AA homozygous variant genotype and A allele showed a significant risk of development of pediatric ALL.

Adenosine Diphosphate-Induced Platelets Aggregability in Polysomnographically Verified Obstructive Sleep Apnea

Obstructive sleep apnea syndrome (OSAS) is a risk factor for arterial thrombosis and cardiovascular morbidity. Activated platelets play key roles in the development of atherothrombosis, thus may be involved in these complications of OSAS. Herein, we evaluated the relationship between severity of OSAS and the degree of platelet aggregates as a marker of activated platelets in 64 patients with OSAS. Platelet aggregations were determined by means of optical aggregometry, using adenosine diphosphate (ADP) as an agonist. Compared with the control group, ADP-induced platelet aggregability was increased in patients with total OSAS, severe OSAS, and in mild to moderate OSAS. Moreover, ADP-induced platelet aggregation was correlated with the Epworth sleepiness scale (ESS) in patients with severe OSAS. Obstructive sleep apnea syndrome is associated with enhanced platelets aggregations, which may predispose the cardiovascular sequels. The ESS may be important in predicting platelet activation and thus atherothrombotic complications in those with OSAS.

Interferon-gamma (IFN-) and Interleukin-2 (IL2) as immunological markers in pulmonary tuberculosis

Early diagnosis and treatment are important in prevention of tuberculosis (TB) infection. World commonly diagnose pulmonary TB using ZN stain, culture and TST. These tests are either low sensitive or required a long time. Recently intracellular interferon gamma flow cytometry assay has been available for diagnosis of pulmonary TB. We assess this diagnostic method in the diagnosis of pulmonary TB with special concern on its role and interleukin-2 (IL2) as tools to observe the effectiveness of anti-tuberculosis therapy. In our study we aimed to evaluate the diagnostic potential of flow cytometry assay for diagnosis of active pulmonary TB, assess the levels of intracellular Interferon-gamma (IFN-γ) and IL2 in patients with pulmonary TB as tools to observe the effectiveness of anti-tuberculosis therapy and correlate between the levels of IFN-γ and IL2 in the patients with the clinical and radiological findings. In our study intracellular interferon gamma flow cytometry assay and IL2 release were evaluated for pulmonary TB patients, LTBI persons and healthy control persons. Flow cytometry were done twice for the pulmonary TB patients, first at the start of treatment and second after 3 months of treatment. We confirmed that the intracellular interferon gamma flow cytometry assay after M. tuberculosis-specific stimulation is sufficient to recognize active TB. IL-2 were more frequently observed in latent TB infected individuals in contrast to active TB patients, and declined with advanced stage of TB. Intracellular interferon gamma flow cytometry assay could function as a powerful immunodiagnostic test to explore pulmonary TB and to predict the efficacy of antituberculosis treatment after 3 months of treatment in cases of pulmonary TB while, IL2 cannot be used to monitor the efficacy of antituberculosis treatment but it declined with advanced stage (stage III) of pulmonary TB in compare to stages I and II. Key words: Intracellular Interferon-gamma (IFN-γ), flow cytometry, pulmonary tuberculosis (TB), interleukin-2 (IL2).

Improved Biological Activities of Isoepoxypteryxin by Biotransformation

Isoepoxypteryxin is the major coumarin of a Japanese medicinal plant Angelica shikokiana. This research was designed to study the effect of structural changes through fungal biotransformation on the reported biological activities of isoepoxypteryxin. Among the tested microorganisms, only Cordyceps sinensis had enzymes that could catalyze the ester hydrolysis and the reductive cleavage of the epoxide ring of isoepoxypteryxin, separately, to give two more polar metabolites (+)‐cis‐khellactone (P1) and a new coumarin derivative (+)‐cis‐3′‐[(2‐methyl‐3‐hydroxybutanoyl)oxy]‐4′‐acetoxy‐3′,4′‐dihydroseselin (P2), respectively. The polar metabolite P2 showed stronger cytotoxicity and higher selectivity than isoepoxypteryxin. On the molecular level, P2 showed more in vitro inhibition of both tubulin polymerization and histone deacetylase 8 (HDAC8). Similarly, P2 showed more neuroprotection against amyloid beta fragment 1 – 42 (Aβ1 – 42)‐induced neurotoxicity in human neuroblastoma cells (SH‐SY5Y) and exhibited more inhibition of the in vitro aggregation of Aβ1 – 42. Both metabolites showed stronger antiplatelet aggregation by increased inhibition of thromboxane‐A2 synthase (TXS) activity and thromboxane‐A2 (TXA2) production. This study is the first to describe the improved cytotoxic, neuroprotective, and antiplatelet aggregation activities of isoepoxypteryxin through its biotransformation by C. sinensis.

Serum Concentrations of Interleukin-33 and its Soluble Receptor sST2 in Patients with Persistent Atrial Fibrillation

Objectives: Interleukin-33 (IL-33) is a new member of the IL-1 cytokine family, which is thought to be involved in the pathogenesis of various inflammatory diseases, through its soluble receptor ST2. There is increasing evidence that inflammation is a relevant player in structural atrial remodeling that represents the main mechanism for atrial fibrillation (AF) persistence. This study was designed to investigate the state of IL-33/ST2 axis serum concentrations in patients with persistent AF. Design and Methods: We investigated the concentrations of IL-33, its soluble ST2 receptors, and high-sensitivity C-reactive protein levels (hsCRP) in the sera of 92 patients with persistent atrial fibrillation, and 68 controls. Results: Serum concentrations of IL-33, sST2, and hsCRP were all significantly elevated in patients with persistent AF compared to controls (P <0.0001 for all). Moreover, serum IL-33 concentrations was positively correlated with the inflammatory marker hsCRP (r=0.606, P =0.002). Conclusion: These preliminary results may support the role of inflammation in AF pathogenesis and IL-33/sST2 axis may be involved in the inflammatory process in AF.