Study of CCR5 Gene polymorphism in rheumatoid arthritis

Chemokines are critical for the inflammatory process in autoimmune diseases such as rheumatoid arthritis [RA]. The chemokine receptor-5 [CCR5] mediates chemotaxis by CC chemokines and is expressed by lymphocytes with the phenotype and monocyte/macrophages. A 32bp deletion in the CCR5 [CCR5-delta 32 allele] abolishes receptor expression in homozygotes, while CCR5-delta 32 carriers express less receptor level than wild type homozygotes. This polymorphism is related to resistance to HIV-1 infection and progression to AIDS. It is hypothesized that CCR5-delta 32 allele may modulate the inflammatory response involved in rheumatoid arthritis and therefore may affect disease severity, susceptibility or both. In the present study 70 rheumatoid arthritis patients and 40 healthy individuals were genotyped. The frequency of CCR5-delta 32 allele was significantly higher in healthy individuals compared to rheumatoid arthritis patients [45% Vs 17%] respectively [p.value 0.033]. Homozygous delta 32 mutation was not detected in patients or controls No significant difference was found between CCR5-delta 32 carriers and wild type homozygotes regarding clinical or laboratory findings except for the tender joint count and rheumatoid factor positivity which was higher in wild type homozygotes [p.value 0.046 and 0.007 respectively]. Our data suggest that CCR5-dlta 32 carriers may partially protected against rheumatoid arthritis, and suggest CCR5 receptor as a candidate for targeted therapy in rheumatoid arthritis

Vascular endothelial growth factor [VEGF-C] signaling through KDR [VEGFR-2] and FLT-4 [VEGFR-3] mediates leukemic cell proliferation and survival

VEGF, a key angiogenic molecule, is a multifunctional cytokine that acts both as a potent inducer of vascular permeability and as a specific endothelial cell mitogen. Because of its effects on endothelial cell growth and microvascular permeability, VEGF is believed to be an important mediator of tumor angiogenesis. Leukemic cells not only release VEGF but also express its receptors, resulting in the establishment of an autocrine loop that supports their migration and survival. VEGF-C may play an important role in the pathophysiology of hematopoietic malignancies by not only regulation of lymphangiogenesis, in vivo, but also by promotion of angiogenesis invasion of neoplastic cells into lymphatic vessels and enhancing lymphatic metastasis during tumor progression. although it is well established that growth in solid tumors is dependent on the formation of neovasculature, the role of angiogenesis in hematopoietic neoplasms has not been determined. The present study was undertaken to identify whether VEGF-C and its receptors VEGFR-2 [KDR] and VEGFR-3 [FLT-4] were expressed in patients with denovo acute leukemia by RT-PCR and to evaluate the relationship between their expression and clinical, laboratory findings and prognosis. Using reverse transcription polymerase chain reaction analysis [RT-PCR], 30 de novo acute leukemia patients [20 ALL patients and 10 AML patients] as well as 10 controls were tested for the expressions of VEGF-C, VEGFR-3 [FLT 4] and VEGFR-2 [KDR] genes. In the current study, VEGF-C, FLT-4 and KDR were detected in 10% of control samples. In ALL patients VEGF C was expressed in 65% of cases, FLT-4 in 70% of cases and KDR in 30% of cases. The expressions of VEGF-C, FLT-4 and KDR in ALL patients were associated with increased risk of leukemia [with OR 16.7 and 95% CI 1.7-160.4, OR 21.0 and 95% CI 2.2-204.6 and with OR 3.9 and 95% CI 0.4-37.6 respectively]. In AML patients, VEGF-C was expressed in 60% of cases, FLT-4 in 70% of cases and KDR in 40% of cases. The expression of VEGF-C, FLT-4 and KDR in AML patients was associated with increased risk of leukemia [with OR 13.5 and 95% CI 1.2-152.2, OR 21.0 and 95% CI 1.8-248.1 and OR 6.0 and 95% CI 0.5-67.7 respectively]. In the 6 followed-up ALL patients, 3 [50%] were in remission, three of them were VEGF-C negative, 2 were FLT4 positive and 1 was KDR positive. 2 of the 6 ALL patients [33.3%] were resistant to treatment, both were VEGF-C positive, FLT-4 positive and 1 was KDR positive. One of the 6 ALL patients [16.6% died during induction, this patient was VEGF-C positive and FLT-4 and KDR negative. In the 4 followed-up AML patients, 3 of them [75%] were in remission, 1 of the 3 was VEGF-C positive and 2 were FLT-4 positive. One of the four AML patients [25%] was resistant to treatment, this patient was VEGF-C, FLT-4 and KDR positive. The number of VEGF-C positive patients with no treatment failure was lower than the number of VEGF-C positive patients with treatment failure. Also, the risk of failed induction was found to be greater in VEGF-C positive patients than in VEGF-C negative patients, thus, the expression of VEGF-C and its receptors [FLT-4 and KDR] in ALL and AML patients was associated with increased risk of leukemia and unfavorable treatment outcome. VEGF-C and its receptors KDR [VEGFR-2] and FLT-4 [VEGFR-3] may play an important role in the pathophysiology of hematopoietic malignancies and may actually contribute to the development of leukemia. Also, owing to the importance of angiogenesis in tumor progression and the effects of VEGF-C, KDR and FLT-4 in chemotherapy-treated leukemias, inhibition of VEGF-C signaling represents an attractive cancer treatment

Assessment of the role of some haemostatic parameters in patients with cryptogenic stroke

Cryptogenic stroke [CS] is a stroke of unexplained aetiology, in 1/3 the of cases, the cause of stroke remains undetermined inspite of full investigations. Patient with CS are thought to have a state of hypercoagulablity. To unmask some of the pathogenic mechanisms underlying cryptogenic stroke through assessment of some genetic disorders including C6[77]T mutation methyl-enetetrahydrofolate reductase gene, activated protein C [APC] resistance and role of thrombin anti-thrombin complex concentration [TAT] in plasma as indicators of hypecoagulable state. The study was conducted on 20 Egyptian patients divided into 2 groups, group I included 10 patients [6 males and 4 females] with cryptogenic stroke aged less than 50 years and group II included 10 age and sex matched patients with non-cryptogenic stroke. All of the 20 cases studied were subjected to panel of investigations including routine laboratory tests and imaging studies in orders to exclude any risk factors for stroke in group I patients and to determine risk factor of stroke in group II. Both groups were investigated for C6[77]T mutation in methylenetetrahydrofolate reductase gene, activated protein C [APC] resistance and thrombin anti-thrombin complex concentration [TAT] in plasma. No statistical significant difference was found between the two groups as regard C6[77]T mutation in methylenetetrahydrofolate reductase gene, [APC] resistance and TAT concentration in plasma [p value >0.05]. However, TAT level was found to be positively correlated with the clinical severity in non-cryptogenic stroke [p value <0.05]. C6[77]T mutation in methylenetetrahydrofolate gene, [APC] resistance and TAT concentration in plasma are not independent risk factors for cryptogenic stroke. TAT could be used as indicator of clinical severity and prognosis in patient with non-cryptogenic stroke

Assessment of genomic damage in medical personnel exposed to ionizing radiation and ultrasound by mean of FISH

Establishing a diagnosis of acute coronary syndrome in the clinical setting remains a challenging task. The advent of testing for cardiac biomarkers such as myoglobin, creatine kinase [CK-MB], and the troponins has facilitated this process. Unfortunately, although these blood markers are extremely sensitive for the identification of patients with myocardial necrosis, their ability to identify patients with acute coronary ischaemia remains limited. During myocardial ischaemia, several changes occur in the amino terminus of ischaemia, several changes occur in the aminot terminus of albumin. Therefore, if reliable, an assay measuring IMA might represent a promising marker for early identification of patients with myocardial ischaemia. To assess the role of IMA and its predictive value for early diagnosis of patients with acute coronary syndromes. Seventy three patients with suspected ACS attending the emergency department at Assiut university hospital were included in addition to -sex and age matched -20 healthy control subjects. All patients were presented within 6h of the typical chest pain episode with negative troponin and normal serum albumin levels. Any patient with liver diseases, renal failure, anaemia, malignancy, acute infections, peripheral vascular diseases, cerebral ischaemia and physical exercise within the last 48 hours was excluded. Full history, clinical examination and standard 12 lead ECG, laboratory investigations including CPK, IMA, CRP, lipogram, kidney and liver functions were done. Twenty two patients were diagnosed as non ischaemic chest pain [NICP] and 51 cases as ACS. CPK and troponin levels were normal in all groups at presentations but 6 hours later CPK levels were significantly higher in ACS patients if compared with NICP or control groups [p<0.000]. on the other hand, IMA levels were statistically significantly high in ACS group only [p<0.000] with a good negative predictive value to diagnose NICP [86.3%]. Moreover, IMA levels were statistically significantly higher in patients finally diagnosed as unstable angina [UA] than those who diagnosed as non ST elevation myocardial infarction [NSTEMI] [p<0.04] meanwhile, it is insignificantly higher than that in STEMI patients. The sensitivity of IMA to predict ACS cases [94.1%] was higher than that of ECG [78.4%] and CPK at presentation [14.7%] and after 6 hours [54.7%]. On the other hand, the specificity of IMA, ECG, CPK at presentation and 6 hours later [86.4%, 90.9%, 86.4% and 97% respectively]. IMA can be used at the emergency setting to exclude the diagnosis of ADS. Moreover, the use of IMA as a diagnostic biomarker in addition to standard markers of myocardial injury is very useful for the evaluation of patients with suspected ACSMedical personnel are without doubt exposed to many biological and chemical agents [e.g., ionizing radiation, ultraviolet light rays, ultrasound, alkylating agents, anesthetic gases and antineoplastic agents] which are efficient inducers of chromosomal aberrations. X-ray and ultrasound are used extensively in many branches of modern medicine. X-rays, together with gamma rays and UV light are all part of the electromagnetic spectrum. Their interaction with biologic material depends on the frequency of wavelength of the radiation. At the short wavelengths of X-rays, electromagnetic radiation has sufficient energy to produce ionization as a result of the removal of electrons from atoms, and are thus called ionizing radiation. Ultrasound [term used to describe mechanical vibrations at frequencies above the human limit of audibility] has experienced phenomenal growth in medical diagnosis in recent years and it is usable in many sensitive circumstances where X-rays might be damaging. The aim of the present study is to assess genomic damage produced by occupational exposure to X-rays and ultrasound equipment using FISH technique using the whole chromosome paint probes for chromosome 1 and 2. Our study revealed chromosomal aberration in 2 personnel exposed to ionizing radiation, continuously for more than 10 years, in the form of absent signaling of chromosomes 2 which denotes total loss [monosomy] of this chromosome. In conclusion, our results may not provide significant proof of the cytogenetic damage caused by occupational exposure to ionizing radiation and ultrasound, however it may prove useful for future human population studies in this field and surely concluded that the necessity of following safety rules in fields dealing with radiations

influence of platelets collagen receptor expression and platelets glycoprotein Ia allele polymorphisms in acute coronary disease

Platelets play an important part in arterial thrombosis therefore it is important to consider the role of adhesion molecules of the platelet surface that play a role in increasing arterial risk. Glycoprofein Ia/IIa is the major platelet collagen receptor and is responsible for platelet adherence to exposed vascular subendothelium. A four fold variation of the platelets receptor density of the collagen receptor glycoprotein Ia/IIa correlating with platelet function to adhere to collagen type I and type III. GPIa/IIa surface expression is influenced by two linked single nucleotide polymorphism [807CT, 873GA] in glycoprotein Ia gene. Individuals with low receptor,densities are homozygos for the 807C/873G allele [CC] genotype whereas individuals homozygos for the 807T/873A allele [TT] genotype have high receptor densities with increased risk of thrombosis. The aim of this study was to evaluate the prevalence of platelet glycoprotein Ia allele polymorphism and platelet collagen receptor [CD49/CD29] density in patients with acute coronary syndrome and control subjects to clarify their possible involvement of their genotype and density as risk factors of the disease, and also to study the association between these findings and the other risk factor for myocardial infarction. The study including 41 patients with a mean age 55.23 years [ +/- 10.98] with a male to female ratio of 33:8. They were compared to 22 controls with a mean age of 49.74 years [ +/- 12.05] and a male to female ratio of 11:11.Glycoprotein Ia gene polymorphism analysis was done by PCR technique for patients and control group together with flowcytometric study of platelet collagen receptor [CD49/CD29] density. Plateletes glycoprotein Ia/IIa receptor% among cases showed a mean of 79.22% [ +/- 12.95] in comparison to 70.9% [ +/- 13.68] among controls. The difference was statistically significant. As regards gene rearrangements, the frequencies of homozygotes T807 allele [TT genotype] were significantly higher in patients with ACS than in controls [24.39% vs 13.63% p<0.05]. The prevalence of [CC] genotypes was also higher in control than in patients [31.8% vs 17%, p<0.05]. Platelets glycoprotein receptor Ia/IIa% [R%] among these groups showed a significant difference between TT cases [mean of 90.5% +/- 3.5], CC cases [mean of 66.5% +/- 5.75] and CT cases [mean of 79.8% +/- 4.82]. Studying other risk factors [obesity, hypertension [HTN], diabetes mellitus [DM] and lipid profile including total cholesterol, low density lipoprotein [LDL] and high density lipoprotein [HDL]] among our group of patients comparing to control subjects, showed no marked significant difference but not for triglycerides [TG] where was significantly higher in our patients compared to control. It was concluded that TT genotype was over presented among patients with acute coronary syndrome compared to healthy controls, whether this genotype is associated with more severe type of coronary heart disease or not this deserve larger scale study