Cytogenetic and molecular analyses of philadelphia chromosome variants in CML [chronic myeloid leukemia] patients from Sindh using karyotyping and RT-PCR

To determine the frequency of Philadelphia chromosome [Ph] and its variants in chronic myeloid leukemia [CML] cases at a tertiary care hospital of Sindh. The study was conducted at the Department of Pathology, Liaquat University of Medical and Health Sciences, Jamshoro and Isra University Hospital, Hyderabad during May-to-September 2014. Bone marrow and peripheral blood samples from a total of 145 diagnosed cases of CML were collected. Cytogenetic analyses were performed using karyotyping as per the International System for Human Cytogenetic Nomenclature guidelines. All karyotypic images were analyzed using the Cytovision software. In order to identify BCR-ABL transcripts, RT-PCR was performed. Statistical analysis of the data was done using SPSS-version-21.0. Of the 145 samples, a total of 133 [91.7%] were positive for the Ph [Ph+] while 12 [8.3%] were negative for the Ph [Ph-]. Of the 133 Ph+ samples, standard karyotypes were noted in 121 [91%], simple variants in 9 [6.7%] and complex variants in 3 [2.3%] of the samples. All the Ph+ samples [n=133] showed BCR-ABL positivity. Of the 12 Ph- samples, a total of 7 [58.3%] were BCR-ABL-positive and 5 [41.6%] were BCR-ABL-negative. Frequency of the Ph was found to be of 90.9% in CML patients using a highly sensitive technique, the RT-PCR. Cytogenetic abnormalities were at a lower frequency. Cytogenetic and molecular studies must be conducted for better management of CML cases. These findings could be very useful in guiding the appropriate therapeutic options for CML patients

Malabsorption of iron as a cause of iron deficiency anemia in postmenopausal women

Malabsorption is one of the causes of iron deficiency anemia in postmenopausal women. The main objective of this study was to access the frequency of malabsorption in iron deficient anemic postmenopausal women. A total of 123 postmenopausal women were enrolled in the study. Of these 123 women, 50 were included as [control group] and 73 patients with comparable severity of anemia were the [patient group]. Two tablets of ferrous sulfate [200 mg/ tablet] along with one tablet of vitamin C [500 mg] were given to all participants. Serum iron levels were determined on samples collected from all participants before and after the administration of ferrous sulfate. Difference between before and after serum iron levels of normal and patients were compared. No change in serum iron between sample one and sample two represented malabsorption. Out of 73, 5 postmenopausal anemic patients showed no change in their serum iron level after the administration of ferrous sulfate. This study shows that frequency of malabsorption of iron in postmenopausal women is 6.8%. Malabsorption should be considered as a prevalent cause of iron deficiency anemia in postmenopausal women. It should be properly diagnosed and iron response should be monitored properly in postmenopausal women with IDA after oral iron therapy. If a postmenopausal woman does not show any response to oral iron therapy, she should be evaluated for iron loss [blood loss and/or malabsorption]. Intravenous route should be used for the administration of iron in these patients

Effect of clopidogrel on platelet CD markers in normal individuals and in patients with untreated type 2 diabetes mellitus

The objective of this study was to evaluate the effect of clopidogrel response in patients with untreated type 2 diabetes mellitus as compared with normal individuals. One hundred and seven subjects i.e. 32 normal and 75 patients with untreated type 2 diabetes mellitus were enrolled in this study. In the first step, normal subjects as well as diabetic patients were selected and tested for various laboratory parameters and platelets flow cytometry. In the second step, an antiplatelet drug [clopidogrel] was administered for 10 days to each individual enrolled in the study. After 10 days blood samples were collected for platelets flow cytometry. CD41 and CD61 did not show any change after the administration of clopidogrel in resting and activated platelets. CD63 and CD62p positivity was increased in normal and in diabetic patients' platelets after activation with ADPbefore clopidogrel. It was decreased in normal resting and ADPstimulated platelets after clopidogrel treatment. CD63 and CD62p positivity in resting and ADP stimulated patients' platelets was also decreased after clopidogrel treatment. The change was, however, not as marked as in normal subjects

Disorders associated with malabsorption of iron: a critical review

Malabsorption is a disorder of the gastrointestinal tract that leads to defective digestion, absorption and transport of important nutrients across the intestinal wall. Small intestine is the major site where most of the nutrients are absorbed. There are three main mechanisms of malabsorption; premucosal, mucosal and postmucosal. Premucosal malabsorption is the inadequate digestion due to improper mixing of gastrointestinal enzymes and bile with chyme. This could be because of surgical resection of the small intestine or a congenital deficiency of the enzymes and bile responsible for digestion e.g. postgastrectomy, chronic pancreatitis, pancreatic cancer, cystic fibrosis, gallstones, cholangitis etc. Mucosal malabsorption occurs in celiac disease, tropical sprue, Crohn's disease etc. Postmucosal condition arises due to impaired nutrients transport e.g. intestinal lymphangiectasia, macroglobulinemia etc. Disorders of malabsorption lead to decreased iron absorption and produce iron deficiency anemia. Using the index terms malabsorption, postgastrectomy, chronic pancreatitis, pancreatic cancer, cystic fibrosis, gallstones, cholangitis, celiac disease, tropical sprue, Crohn's disease intestinal lymphangiectasia, macroglobulinemia and iron deficiency anemia the MEDLINE and EMBASE databases were searched. Additional data sources included bibliographies and references of identified articles

Comparative analysis of serum iron, serum ferritin and red cell folate levels among breast fed, fortified milk and cow’s milk fed infants

Objective: Iron and folic acid are essential nutrients needed for hematopoiesis. Infants' diet is commonly deficient in these micronutrients that lead to nutritional anemia. Aim of this study was to determine serum iron, serum ferritin and red cell folate levels among healthy breast fed, fortified milk and cow's milk fed infants. Methods: A total of 120 infants of 4-9 months of age were enrolled in this study. It included 40 normal breast fed controls, 40 fortified milk fed [FM] and 40 cow's milk fed [CM] infants. Serum iron, serum ferritin and red cell folate concentrations were determined using colorimetric and enzyme immunoassay techniques. Results: Mean serum iron, serum ferritin and red cell folate concentrations of breast fed control group were 120.9+/-68.4µg/dl, 109+/-71.7ng/ml and 1044.1+/-409.2ng/ml respectively. Fortified milk [FM] group showed significantly decreased serum iron [p<0.003] as compared with controls whereas serum ferritin and red cell folate values showed insignificant change [p=0.25 and p=0.85 respectively]. However serum iron, serum ferritin and red cell folate were significantly decreased in cow's milk fed [CM] group as compared with control subjects [p<0.04, p<0.006, p<0.02 respectively]. Comparison of these biochemical parameters between FM and CM groups showed statistically significant difference of serum ferritin and red cell folate among cow's milk group [p<0.0001 and p<0.02] whereas serum iron level showed no significant difference, a p-value being 0.38. Conclusion: Healthy breast fed infants do not need any supplementation and fortification of iron and folic acid. Fortified milk appears to be an acceptable alternative in the absence of breast milk whereas cow's milk is a poor source of iron and folic acid in infants

Frequency of ABH secretors and non secretors: a cross sectional study in Karachi

ABO blood group and secretor status is valuable in relation to some diseases in clinical and forensic medicine. Across the globe there are geographic and racial differences in the frequency of secretors and non-secretors. Aim of this study was to evaluate the status of ABH blood group secretors and non-secretors in Karachi [Pakistan]. Blood and saliva samples were randomly collected from one hundred and one [n=101] healthy adult students [76 male, 25 female] ranging in age from 15 to 40 years. Their ABO and Rhesus blood groups were determined by conventional methods, and their secretor status was studied by hemagglutination inhibition method of saliva. Results showed that 64.4% of the study population were ABH blood group secretors while 35.6% were non-secretors. Frequencies of the secretor status among various ABO blood groups were 71.4% in group A, 79.5% in group B, 45.5% in group AB, and 61.5% in group O. Frequency of ABH secretor is high [64.4%]. Blood group B has the highest secretor [79.5%] frequency while Blood group AB has the lowest [45.5%]

Platelet receptors: an instrumental of platelet physiology

Platelets play an important role in hemostasis, inflammation, host defense, tumor growth and metastasis. Platelets receptors are instrumental in platelet-platelet aggregation and interaction of platelets with leukocytes, endothelial cells and coagulation factors. These receptors are also the targets for antiplatelet drugs. This review focuses on the role of platelet receptors in human physiology. Data were extracted from peer-reviewed journals using MEDLINE and EMBASE databases, and the following terms [platelets, platelet receptors, CD markers, integrins, tetraspanins, transmembrane receptors, prostaglandin receptors, immunoglobulin superfamily receptors] were used

New horizons in platelets flow cytometry

Platelet flow cytometry is an emerging tool in diagnostic and therapeutic hematology. It is eminently suited to study the expression of platelet surface receptors both qualitatively as well as quantitatively. It can serve as a useful marker for the documentation of in vivo platelet activation, and thus, fore-warn the risk of thromboembolism in patients with diabetes mellitus, coronary syndromes, peripheral vascular diseases, and pre-eclampsia. This technique can also be extended to study and compare the effect of various antiplatelet drugs on the level of activation of platelets and to establish any dose-effect relationship of these drugs. Topographical localization of platelet granules and study of platelet-platelet and platelet-leukocyte interaction is also possible by this procedure. All these parameters serve as pointers towards the presence of activated platelets in the circulation with its thromboembolic consequences. This is a simple reliable and cost effective technique which has a wide application in the diagnosis of various inherited and acquired platelet disorders. Study of platelet cluster of differentiation (CD) markers in various inherited disorders i.e. Bernard Soulier’s disease, von Willebrand disease, Glanzman’s disease, and Grey platelet syndrome may help categories the molecular lesions in these oft under-studied disorders.

Flow cytometry based chronologic analysis of surface glycoproteins of resting and stimulated platelets in normal individuals

To evaluate the effect of timed incubation on the activation dependent glycoproteins on the surface of unfixed platelets after stimulation with adenosine diphosphate [ADP]. A total of 32 normal subjects were recruited in this study. CBC, Hb A1C, fasting and random blood sugar, urea, creatinine, cholesterol, triglycerides, high density as well as low density lipoproteins were determined. Flow cytometric analysis of platelets was done using FACSCalibur [BD Bioscience]. Two sets of CD markers i.e. platelets CD41and CD61 and CD63 and CD 62p were studied during resting state and after stimulation with ADP. Mean +/- SD values of CD41 and CD61 surface markers of resting and stimulated platelets showed no statistical difference; p value being 0.198 and 0.486 respectively. Comparison of CD41 and CD61 markers at 20 minutes Vs 2 hrs, 20 minutes Vs 3 hrs and 2 hrs Vs 3 hrs didn't show any significant difference after stimulation with ADP. This indicates that these markers remain stable and they are not affected by incubation for up to 3 hours. Activation of platelets with ADP resulted in an increase in the number of CD63 and CD62p positive platelets with a p value of 0.001. Activity of CD63 remained high while that of CD62p progressively decreased after incubation for 2 and 3 hours. CD63 is an ideal marker to evaluate functional status of the platelets while CD62p positivity, though it increases after activation, cannot be used alone as a marker of platelet activation because its positivity decreases with the passage of time. Changes in CD62p positivity when considered in conjunction with increased CD63 positivity lend support to the presence of increased number of activated platelets in the peripheral blood

Soluble transferrin receptors in Malaria

To determine the concentration of soluble transferrin receptors [sTfR] in patients with malaria. Cross-sectional, analytical study. Baqai Institute of Haematology, Baqai Medical University, Karachi, from December 2009 to April 2010. Twenty samples from normal male and female subjects each were drawn for establishing the reference range while 38 from patients with malaria [with or without anaemia] sTfR centration was determined. Descriptive statistics were used for data analysis. Out of 38 patients, 4 had iron deficiency anaemia while 34 patients were without anaemia. Mean sTfR level in the control group was 33.53 +/- 4.38 nmol/l. In patients with malaria without iron deficiency anaemia, mean sTfR concentration was 30.84 +/- 5.40 nmol/l. Patients with malaria and concomitant iron deficiency anaemia had mean sTfR level of 101.67 +/- 11.69 nmol/l. Comparison of sTfR in normal subjects and in patients with malaria showed no statistically significant difference [p = 0.208]. Statistically significant difference [p < 0.001] was observed in patients with malaria and concomitant IDA as compared to normal control group. Malaria without concomitant iron deficiency anaemia had near normal sTfR levels. While those with concomitant iron deficiency anaemia had significant higher level of sTfR. This concludes that these receptors are not affected in malaria alone

Soluble transferrin receptors in the diagnosis of iron deficiency anemia, beta thalassemia minor and beta thalassemia minor with concomitant iron deficiency anemia

Iron deficiency anemia and beta thalassemia minor are two important causes of microcytic and hypochromic anemia, beta thalassemia minor with concomitant iron deficiency anemia make the diagnosis difficult through conventional laboratory tests. Determination of soluble transferrin receptors is a helpful laboratory test for the diagnosis with certainty. Purpose of this study was to evaluate the role of soluble transferrin receptors in the differentiation of iron deficiency anemia from beta thalassemia minor. A total of 80 subjects were enrolled in this study. They were divided into four groups i.e. 20 normal adult male, 20 normal adult female, 20 patients with iron deficiencyanemia group and 20 patients with beta thalassemia minor. Patients with beta thalassemia minor were further sub grouped in beta thalassemia minor with and without concomitant iron deficiency anemia. Soluble transferrin receptors were determined by ELISA technique using Quantikine IVD kit [R and D Systems]. Levels of sTfR in individuals with beta thalassemia minor were increased but these were lower than in iron deficiency anemia group. Mean sTfR levels were higher in patients with beta thalassemia minor and concomitant iron deficiency anemia than in normal subjects as well as in patients with beta thalassemia minor alone. Their levels were similar to those in patients with iron deficiency anemia. sTfR can be used as a discriminating marker between patients with iron deficiency anemia and beta thalassemia minor alone. Care must be taken while dealing the patients with beta thalassemia minor and concomitant iron deficiency anemia

Soluble transferrin receptor: a differentiating marker between iron deficiency anaemia and anaemia of chronic disorders

Background: Iron deficiency anaemia and anaemia of chronic disorders are the two major causes of microcytic and hypochromic anaemia. Many times the diagnosis of these conditions becomes difficult through conventional laboratory tests. Determination of soluble transferrin receptors is a helpful laboratory test for the differential diagnosis of these conditions. The study was conducted to evaluate the role of soluble transferrin receptors in the differential diagnosis between iron deficiency anaemia and anaemia of chronic disorders. Methods: A total of 80 blood samples were evaluated, i.e., 20 samples from normal adult male, 20 samples from normal adult female, 20 samples from iron deficiency anaemia group and 20 samples from patients with anaemia of chronic disorders. Soluble transferrin receptors were determined by ELISA technique using Quantikine IVD kit [R and D Systems]. Results: There was significant difference in the levels of sTfR in iron deficiency anaemia and anaemia of chronic disorders. Statistically non-significant difference was observed between the levels of sTfR in patients with anaemia of chronic disorders as compared to normal control group. Conclusion: The sTfR determination can be used as a reliable differentiating marker in the diagnosis of iron deficiency anaemia and anaemia of chronic disorders. Keywords: sTfR, iron deficiency anaemia, anaemia of chronic disorders