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Article
Evaluation of (HPLC) Patterns of Sickle Cell Anaemia Patients in Comparison with Apparently Healthy Individuals

Author: Zuhair Mohammad Ali Jeddoa
Journal: Karbala Journal of Medicine مجلة كربلاء الطبية ISSN: 19905483 Year: 2011 Volume: 4 no 1 Issue: 9 Pages: 980-987
Publisher: Kerbala University جامعة كربلاء

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Abstract

background: Sickle cell haemoglobin (HbS) results from an autosomal recessively inherited mutation in which the amino-acid glutamine is replaced by valine at position 6 in the beta globin chain of haemoglobin (Hb). Sickle cells have a reduced deformability and are easily destroyed, causing occlusion of the microcirculation and a chronic haemolytic anaemia with a median Hb concentration level of about 9 g/dl. Routine electrophoresis methods and High performance liquid chromatography (HPLC) were used to screen normal and variant Hb, and allowed the verification of the Hb observed with electrophoresis and precise quantification of their proportion.Objectives: 1- This study aimed to evaluate the chromatographic pattern of Hb types (HbA, HbF, HbA2 and HbS) of sickle cell anemia patients in comparison with the apparently healthy individuals.2- To study the Hb chromatographic patterns according to the gender, age and blood groups.3- To evaluate the efficiency of variant Hb testing system in detection of HbS type of sickle cell anaemia patients.Methods: A total of eighty four sickle cell anemia patients who were attending to theIbn Al-Baladi pediatrics hospital (in Baghdad) and Kerbala teaching pediatricshospital, the samples were (39) males and (45) females , from April of 2006 toFebruary of 2007 . And thirty seven of case controls with matched age and sex wererandomly selected from apparently healthy individuals. High performance liquidchromatography (HPLC) was adopted to determine the different types of Hb forpatients and control groups using variant Hb testing system which depend upon theseparation and quantification of Hb types by high performance liquid chromatographytechnique.Results: The study of Hb chromatographic patterns of samples revealed that there were no significant differences (p ≥ 0.05) between the values of HbA2 for the patients and control groups and there were significant differences (P≤ 0.05) for HbF and highly significant differences (P≤ 0.01) for HbA and HbS percentages of patients in comparison with the control group. The results of Hb chromatographic patterns of samples according to the gender revealed that there were no significant differences at (p≥ 0.05) between males and females within patient and control groups. The results of Hb chromatographic patterns of Hb types for patients and control groups according to the age groups revealed heterogeneous results with highest HbF, HbA, and HbA2 percentages of patients were (10.83±3.32, 57.6±7.33, and 4.22 ±1.88) respectively in age group less than five years old, while it was (54.33±8.9) for HbS type in age group (6-10) years old. As for control group, the highest HbF percentages was (8.2±4.09) in age group less than five years old, for HbA was (87.22±5.86) in age group (6-10) years old, for HbA2 was (3.6±0.23) in age group (11-15) years old, and for HbS was (0.3±0.05) in age group less than five years old. Finally, the results also showed that there were no significant differences at (P ≥ 0.05) for Hb chromatographic patterns of different Hb types percentages according to the blood groups of studied samples within group (patients or controls).Conclusion:1- The study of Hb chromatographic patterns is useful for the diagnosis of sickle cell anaemia.2- There are no significant effects of gender and blood groups on the chromatographic patterns of different Hb types of sickle cell anaemia patients in comparison to the apparently healthy individuals.3- HPLC is an excellent, powerful diagnostic tool for the direct identification of HbS.


Article
Homocysteine ,Folic acid ,Vitamin B12 and Pyridoxine : Effects on Vaso-Occlusive Crisis in Sickle Cell Anemia and Sickle –Thalassemia

Authors: Sawsan Sati' Abbas --- Najat Abul –Razak --- Noor Mustafa --- Razeqa Abd Ali
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2011 Volume: 10 Issue: 4 Pages: 473-479
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

ABSTRACT:BACKGROUND:Hemoglubiopathies include sickle cell anemia and the Thalassemia . S/ β0- Thalassemia have a clinical course similar to HbSS. Elevated concentration of Homocysteine contribute to thrombosis , a frequent event in sickle cell anemia . Vitamin B12, Pyridoxine , and folic acid deficiencies lead to dangerous increase in plasma Homocysteine.OBJECTIVE:1- To test whether children with sickle cell anemia and Sickle cell – Thalassemia have elevated concentration of serum Homocysteine with diminished level of folate, B6, B12.2- To determine whether hyperomocysteinaemia has a correlation with the frequency of Vaso-occlusive crisis.PATIENTS AND METHOD:A case- control study was carried over a period of one year from Jan.- Dec. 2010 inclusive, 30 patients were collected from the Thalassemia centre in Ibn –AL-Baldy Hospital together with healthy 30 cases, age and sex matched ,were taken from AL- Kadhimiyia Teaching Hospital. Venous blood sample were aspirated from both groups to estimate serum Homocysteine, Folic acid, B12 and B6 level. Statistical analysis was done, using the student T-test ( P. value < 0.05 is considered as statistically significant) . Pearson correlation analysis was performed.RESULTS :The age of the patients range between (5-29) years, the majority of the patients were between ( 10 -19 ) years , 10 cases ( 38 .46 %). More than one half were male ,16 cases ( 61.54 %) . Sickle cell – Thalassemia constitute 20 cases (76.93%). Vaso-occlusive crisis was mainly involving the large joints, 15 cases (57.69 %). Mild attacks constitute more than half of the patients, 16 cases (61.54 %).Homocysteine level was higher in the patients group compared with control group with a mean and standard deviation of (44.52 ± 23.008) and (18.65 ± 4.56)μmol/L respectively . Folic acid level was lower, B12 level was higher, B6 level was lower in the patients group compared with control group with a mean and standard deviation of (11.32± 3.23) and (14.71 ± 3.39)ng /ml, (172.57± 61.34) and (103.45 ± 30.45)pg /ml, (4.43± 3.93) and (10.23 ± 2.30) ng/ml respectively, the results were statistically not significant, P. value > 0.05. Significant inverse correlation was found between Homocysteine level and B6 level. A strong positive correlation between Homocysteine level and the frequency of Vaso-occlusive crisis was found.CONCLUSION:Patients with sickle cell disease have high serum level of Homocysteine with low level of folic acid and pyridoxine. This Hyperomocysteinaemia is significantly inversely correlated with pyridoxine deficiency , but positively correlated with the frequency of Vaso-occlusive crisis.


Article
Homocysteine ,Folic acid ,Vitamin B12 and Pyridoxine : Effects on Vaso-Occlusive Crisis in Sickle Cell Anemia and Sickle –Thalassemia

Authors: Sawsan Sati' --- Najat Abul –Razak --- Noor Mustafa --- Razeqa Abd Ali
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2010 Volume: 9 Issue: 4 Pages: 473-479
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

ABSTRACT:BACKGROUND:Hemoglubiopathies include sickle cell anemia and the Thalassemia . S/ β0- Thalassemia have aclinical course similar to HbSS. Elevated concentration of Homocysteine contribute to thrombosis , afrequent event in sickle cell anemia . Vitamin B12, Pyridoxine , and folic acid deficiencies lead todangerous increase in plasma Homocysteine.OBJECTIVE:1- To test whether children with sickle cell anemia and Sickle cell – Thalassemia have elevatedconcentration of serum Homocysteine with diminished level of folate, B6, B12.2- To determine whether hyperomocysteinaemia has a correlation with the frequency of Vasoocclusivecrisis.PATIENTS AND METHOD:A case- control study was carried over a period of one year from Jan.- Dec. 2010 inclusive, 30patients were collected from the Thalassemia centre in Ibn –AL-Baldy Hospital together withhealthy 30 cases, age and sex matched ,were taken from AL- Kadhimiyia Teaching Hospital.Venous blood sample were aspirated from both groups to estimate serum Homocysteine, Folic acid,B12 and B6 level. Statistical analysis was done, using the student T-test ( P. value < 0.05 is consideredas statistically significant) . Pearson correlation analysis was performed.RESULTS :The age of the patients range between (5-29) years, the majority of the patients were between ( 10 -19 ) years , 10 cases ( 38 .46 %). More than one half were male ,16 cases ( 61.54 %) . Sickle cell –Thalassemia constitute 20 cases (76.93%). Vaso-occlusive crisis was mainly involving the largejoints, 15 cases (57.69 %). Mild attacks constitute more than half of the patients, 16 cases (61.54%).Homocysteine level was higher in the patients group compared with control group with a meanand standard deviation of (44.52 ± 23.008) and (18.65 ± 4.56)μmol/L respectively . Folic acid levelwas lower, B12 level was higher, B6 level was lower in the patients group compared with controlgroup with a mean and standard deviation of (11.32± 3.23) and (14.71 ± 3.39)ng /ml, (172.57±61.34) and (103.45 ± 30.45)pg /ml, (4.43± 3.93) and (10.23 ± 2.30) ng/ml respectively, the resultswere statistically not significant, P. value > 0.05. Significant inverse correlation was found betweenHomocysteine level and B6 level. A strong positive correlation between Homocysteine level and thefrequency of Vaso-occlusive crisis was found.CONCLUSION:Patients with sickle cell disease have high serum level of Homocysteine with low level of folic acidand pyridoxine. This Hyperomocysteinaemia is significantly inversely correlated with pyridoxinedeficiency , but positively correlated with the frequency of Vaso-occlusive crisis

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