Detection of BCR-ABL1 fusion gene transcripts in the saliva of Nigerian patients with chronic myeloid leukemia.

BACKGROUND: The presence of BCR-ABL1 fusion gene resulting from a t(9; 22) reciprocal chromosome translocation is the molecular hallmark of chronic myeloid leukemia (CML). In the diagnosis and treatment of CML, peripheral blood or bone marrow samples are usually taken for analysis. However, both methods are invasive sample collection methods, thus a noninvasive saliva sample method for the detection of the fusion gene transcripts (BCR-ABL) was investigated in some Nigerians with CML. MATERIALS AND METHODS: Real-time (RT)-polymerase chain reaction (PCR) analysis was used to detect BCR-ABL1 fusion gene in the saliva and blood of 42 Nigerian CML patients. RNA was extracted using RNeasy kit and reverse transcribed by random hexamer priming using murine Moloney reverse transcriptase. BCR-ABL1 transcript types were first detected by multiplex PCR and then quantified by a duplex RT-PCR-TaqMan chemistry with MGB probe and Black Hole Quencher. RESULTS: Of the 42 subjects, transcript types were detected in 36 (85.7%) samples, e13a2 fusion transcript sub-type was detected in 9 (21.4%), whereas e14a2 subtype was found in 27 (67.3%); six (14.3%) of the samples did not reveal any of the fusion transcript subtypes. The median BCR-ABL1 messenger RNA values were 9.38 × 102 in saliva and 10.29 × 104 in blood (P < 0.05). Similarly, the median ABL1 value in saliva (3.11 × 103) was significantly lower (P < 0.01) than in blood (4.22 × 103). However, the median BCR-ABL1 ratio in saliva (14.5%) was not significantly different (P = 0.8) from that of blood (12.0%). CONCLUSION: Saliva may offer an alternative easy-to-collect, readily available, and noninvasive sample for the diagnosis and treatment of CML.

Molecular characterization of exon 28 of von Willebrand's factor gene in Nigerian population.

BACKGROUND: Polymorphisms in von Willebrand factor (VWF) gene are an important contributor to the expression of VWF gene and differences in ethnic distribution of these single nucleotide polymorphisms (SNPs) exists. AIMS: Our objective was to molecularly characterize the exon 28 of the VWF gene in the three major ethnic groups of Nigeria. SUBJECTS AND METHODS: We recruited 90 subjects, 45 had a history of bleeding. Questions included those used in the Zimmerman Program for the Molecular and Clinical Biology of von Willebrand disease (VWD), and the bleeding scores were calculated using the Molecular and Clinical Markers for the Diagnosis and Management of type 1 VWD scoring system. Full blood count, coagulation profile, VWF:antigen level and VWF:collagen-binding activities were carried out. Data were analyzed using GraphPad Prism (5.03). GraphPad Software, Inc USA. The BigDye terminator chemistry was used to determine the nucleotide sequences of VWF gene (exon 28). RESULTS: Eight SNPs were identified, rs 216310 (T1547), rs 1800385 (V1565L), rs1800384 (A1515), rs1800383 (D1472H), rs 1800386 (Y1584C), rs 216311 (T1381A), rs 216312 (intronic) and rs 1800381 (P1337). CONCLUSION: The SNPs rs 216311, rs 1800383 and rs 1800386 associated significantly with bleeding in study subjects. rs1800386 occurred in all with bleeding history, no ethnic variations were noted.

The effect of potassium bromate on some haematological parameters of Wistar rats.

Potassium bromate used widely in foods has been associated with various complications in humans. However there is paucity of literature on adverse effects on haematological parameters. Thus we decided to carry out an experimental study to determine the effects of potassium bromate on some blood indices using Wistar rats. Twenty (20) male Wistar rats aged 2-3 months obtained from the department of Veterinary Medicine, University of Nigeria Nsukka were acclimatized for two weeks. They were divided into five groups and fed with graded doses of potassium bromate solution for six weeks. One of the groups served as the control. Pre and post administration blood samples were collected and analyzed the same day using standard methods. The results revealed significant [P<0.05] decrease in the platelet count when compared with the controls. There were no statistically significant differences in the mean cell haemoglobin concentration, haematocrit, and total leukocyte counts between the test and control rats. Potassium bromate is injurious to health if consumed continuously and in high quantity. It has been shown to reduce platelet count in rats, and thus may cause thrombocytopenia in humans. It is therefore imperative to take adequate measures to eliminate the use of potassium bromate in the preparation of food products.

Prothrombin time, activated partial thromboplastin time and other haematological parameters among non-diabetic hypertensive patients.

OBJECTIVES: To screen non-diabetic hypertensive Nigerian patients with a view to determining their haematological and coagulation profiles. SUBJECTS AND METHODS: Thirty (30) consecutive non-diabetic hypertensive patients, aged 20-60 years and 30 age and sex-matched, apparently healthy subjects, were investigated for some haematological and coagulation parameters, blood pressure, pulse, body weight, height and body mass index. RESULTS: There were statistically significant differences (p < 0.05), in some haematological parameters, blood pressure, body mass index and height in the total number of patients studied when compared with the total control group. The separate genders each showed statistically significant differences in both haematological parameters and blood pressure (P < 0.05) when compared with their corresponding control groups. In addition, haematological values, body mass index and height showed statistically significant differences (p < 0.05) when the male and female patients were compared. Overall, 25 (83.33%) of the patient's blood films were normochromic and normocytic. CONCLUSION: It appears that hypertension has adverse effects on haematological parameters. We therefore recommended that coagulations studies should be included in the investigation work-up of hypertensive patients.

Haematological assessment of occupational exposure to lead handlers in Enugu urban, Enugu State, Nigeria.

OBJECTIVE: To determine blood levels of lead and its effects on haematological parameters among occupational lead handlers in Enugu, Enugu State, Nigeria. In developing countries, rapid industrialisation has lead to an alarming demand for lead. Furthermore, the burden of lead toxicity is greatly underestimated. Hence, the need to assess the unavoidable toxic effects of lead as done in this study. METHODS: Blood lead levels were measured by atomic absorption spectrometry (AAS) in eighty one (81) male subjects from three manufacturing companies, all located in Enugu metropolis, Nigeria. Thirty (30) staff of the industries not directly involved in lead handling served as control group I, while twenty (20) apparently healthy individuals from within the same locality not involved in lead handling served as control group II. Haematological values, blood lead levels and blood pressure (BP) were established using standard procedures. Statistical Analysis System (SAS) software was used to analyze the results. P value of < 0.05 was taken as significant. RESULTS: Mean blood levels were 7.00 +/- 0.07 microg/dl in test subjects; 3.00 +/- 0.19 microg/dl in control group I and 2.00 +/- 0.04 microg/dl in the control group II. There were significant statistical differences (p < 0.05 for each) in haemoglobin (Hb), packed cell volume (PCV), reticulocyte, total white blood cell (WBC), monocyte, autohaemolysis without glucose, and systolic and diastolic pressure between subjects and control group I. There were also significant differences (p < 0.05 for each) in the mean levels of Hb, PCV, reticulocyte, eosinophil, monocytes and systolic and diastolic pressures between the test subjects and control group II. There were however, no statistically significant differences (p > 0.05) in the means of other parameters. Basophilic stipplings were not observed in the red cells of those directly exposed to lead. CONCLUSIONS: It is suggested, therefore, that comprehensive and preventive measures towards exposure to lead in work places, and routine haemotological investigations be included in the bio-monitoring of the health status of lead workers.

Biochemical and haematological assessment of workers exposed to some petroleum products in Rnugu Urban, Rnugu State, Nigeria.

BACKGROUND: Exposure to petroleum product(s) occurs during extraction, refining, transportation and utilisation. Surprisingly, this relatively common hazard has received little public recognition. We therefore studied the adverse effects of petroleum products among occupationally exposed liquefied petroleum gas and histochemical workers in Enugu Urban, Nigeria. METHODS: Eight industries/departments were surveyed. Sixty workers were tested for some biochemical and haematological parameters (phosphatases, transaminases, complete blood count, erythrocyte sedimentation rate), were analysed, using standard methods. Also, 30 age and sex-matched apparently healthy subjects served as the control group. RESULTS: When the test and control results were compared, there were statistically significant differences (p < 0.05) in: (1) haematocrit, reticulocytes, erythrocyte sedimentation rate, platelet, lymphocyte, monocyte, alkaline phosphatase, aspartate transaminase, alanine transaminase among liquefied petroleum gas workers and (2) haemoglobin, haematocrit, erythrocyte sedimentation rate, platelets, neutrophil, lymphocyte, monocytes, acid phosphatases and alkaline phosphatase among histochemical workers. Overall, there was statistically significant decrease (p < 0.05) in most of the parameters among the test subjects when compared with the control group. Blood films of the workers revealed mild to moderate hypochromia, anisocytosis and poikilocytosis while the controls were normocytic and normochromic. CONCLUSION: We conclude that exposure to petroleum products adversely affects biochemical and haematological parameters. Hence, improved working conditions are recommended.

Biochemical and haematological assessment of workers exposed to some petroleum products in Enugu Urban, Enugu State, Nigeria.

BACKGROUND: Exposure to petroleum product(s) occurs during extraction, refining, transportation and utilisation. Surprisingly, this relatively common hazard has received little public recognition. We therefore studied the adverse effects of petroleum products among occupationally exposed liquefied petroleum gas and histochemical workers in Enugu Urban, Nigeria. METHODS: Eight industries/departments were surveyed. Sixty workers were tested for some biochemical and haematological parameters (phosphatases, transaminases, complete blood count, erythrocyte sedimentation rate), were analysed, using standard methods. Also, 30 age and sex-matched apparently healthy subjects served as the control group. RESULTS: When the test and control results were compared, there were statistically significant differences (p < 0.05) in: (1) haematocrit, reticulocytes, erythrocyte sedimentation rate, platelet, lymphocyte, monocyte, alkaline phosphatase, aspartate transaminase, alanine transaminase among liquefied petroleum gas workers and (2) haemoglobin, haematocrit, erythrocyte sedimentation rate, platelets, neutrophil, lymphocyte, monocytes, acid phosphatases and alkaline phosphatase among histochemical workers. Overall, there was statistically significant decrease (p < 0.05) in most of the parameters among the test subjects when compared with the control group. Blood films of the workers revealed mild to moderate hypochromia, anisocytosis and poikilocytosis while the controls were normocytic and normochromic. CONCLUSION: We conclude that exposure to petroleum products adversely affects biochemical and haematological parameters. Hence, improved working conditions are recommended.

Association between ABO, Rhesus blood group systems and haemoglobin genotype among confirmed HIV/AIDS-TB co-infected patients in Enugu Urban, Nigeria.

BACKGROUND: The distribution of ABO, Rhesus blood group and haemoglobin (Hb) genotypes was investigated among 320 confirmed human immunodeficiency virus I & II (HIV)/acquired immunodeficiency syndrome (AIDS) with tuberculosis (TB) co-infected patients. One Hundred and Sixty (160) healthy HIV I & II negative age and sex-matched population were used as controls for this study. STUDY DESIGN: Patients and controls were from the same environment and their blood groups and Hb genotypes were determined by the standard tube and electrophoresis methods respectively. RESULTS: The statistical analysis of the results revealed statistically significant association (P<0.01) between Hb-genotype among female patients and female controls chi-squared = 6.099, P<0.01, and also between patient and control groups of both sexes chi-squared = 7.4561, p<0.01 only. CONCLUSION: We conclude that HIV I & II/AIDs with TB co-infection among Nigerians so far studied appear to show no association in the distribution of ABO, Rhesus blood group and Hb-genotype prevalence. Hence they cannot be said to have either protective nor predisposing characteristics.

Evidence that circulating lymphocytes act as vehicles or viraemia in measles.

Previously described giant multinucleated cells were observed in phytohaemagglutinin (PHA) cultures of leucocytes from blood samples of 17 (36%) out of 47 children with measles or undiagnosed fever in Ibadan. The giant cells were more readily seen in PHA cultures of purified lymphocyte preparations than in total leucocyte suspensions. It was concluded that the observation indicates in vivo infection of some blood lymphocytes of patients with measles and related infections by syncitium producing virus to manifest the giant cells seen in culture. The practical application of lymphocyte cultures to the laboratory diagnosis of measles is stressed and the concept of circulating lymphocytes contributing to the systemic spread of measles infection is highlighted.

Blood lymphocytes and measles viraemia.

Previously described giant multinucleated cells were observed in phytohaemagglutinin (PHA) cultures of washed leucocytes from blood samples of seventeen (36%) out of forty-seven children with measles or undiagnosed fever in Ibadan. The giant cells were more readily seen in PHA cultures of purified lymphocyte preparations than in total leucocyte suspensions. It was concluded that the giant cells seen in cultures indicate in-vivo infection of some blood lymphocytes of patients with measles and related infections by syncytium-producing virus. The practical application of lymphocyte cultures to the laboratory diagnosis of measles is stressed, and the concept of circulating lymphocytes acting as vehicles for the systemic spread of measles infection is highlighted.

Chronic lymphocytic leukaemia with raised serum IgM levels.

Eight patients diagnosed as having chronic lymphocytic leukaemia (CLL) from clinical and haematological data were also observed to possess immunological characteristics similar to tropical splenomegaly syndrome (TSS). We suggest that the raised IgM level in these cases may be the effect of chronic exposure to malaria in patients who later developed CLL. Consequently, raised serum IgM, which is a feature of TSS, may also be found in some cases of CLL in Nigeria.

Immunological identification of peripheral-blood lymphocytes from patients with lymphoproliferative disorders in Nigeria. I. Acute lymphocytic leukaemia (ALL).

The peripheral-blood lymphocytes of eight consecutively diagnosed cases of acute lymphocytic leukaemia (ALL) in Nigerians seen over a period of 2 years were classified as T, B and Null cells, using mainly sheep-rosetting (for T cells) and EAC-rosetting (for B cells) techniques. The patients' lymphocytes response to stimulation with phytohaemagglutinin (PHA) and basic haematological parameters were also determined. There were three male cases (37.5%), aged 10, 14 and 21 years who had T-cell ALL and one boy (12.5%) aged 10 years with B-cell ALL. The remaining four patients (50%) were two girls aged 6 and 6 1/2 years and two males aged 10 and 21 years who had lymphocytic leukaemia of the Null-cell class. These results from a tropical environment agree with previous findings among Caucasians with respect to the age and sex incidence and predominance of ALL with Null cells.

Immunological identification of peripheral-blood lymphocytes from patients with lymphoproliferative disorders in Nigeria. II. Chronic lymphocytic leukaemia (CLL).

The surface markers for blood T and B lymphocytes in sixteen consecutively diagnosed patients with CLL were sought for. There were twelve females and four males, and their ages ranged from 30 to 70 years. Although the absolute numbers of T and B lymphocytes were increased, the percentage scores as well as mitogenic responses of these leukaemic cells were depressed, and all the patients except one who has T-cell CLL were considered to have Null-cell CLL. This finding suggests that CLL may take an unusual form in a tropical environment, and this may account for the long remissions which often occur.

Polycythaemia rubra vera associated with unbalanced expression of the X chromosome and monoclonality of T lymphocytes.

Polycythaemia rubra vera (PRV) was diagnosed in a 69-year-old Nigerian woman whose haemolysate revealed an electrophoretically slow-moving homogeneous band of the enzyme glucose 6-phosphate dehydrogenase (G6PD). Further biochemical characterization identified the enzyme as a new sporadic G6PD variant. Electrophoresis of the haemolysate of 4 children of the proposita identified in a daughter a large quantity of the slow-moving variant enzyme, and a much lower quantity of the normal type enzyme, thus, signifying an extremely unbalanced mosaic phenotype situation. All other family subjects studied, including 2 sons, had the normal enzyme type B, thus confirming the heterozygosity of the proposita at the G6PD locus. Homogenates of platelets, granulocytes and E-rosette-forming lymphocytes of the proposita showed an identical slow-moving band as the haemolysate, thus, suggesting the origin of these cells from a common progenitor cell which may be either the pluripotent stem cell (PSC) or another precursor cell at an earlier level of development than the PSC. While the observation of preferential production of the variant enzyme may be attributed to the presence on the X chromosome of a 'haemopoietic gene', its relevance to the etiology of PRV is unclear.

Childhood acute leukaemia in a tropical population.

The clinical features of acute leukaemia (AL) were documented prospectively among Nigerian children resident in the South-Western rain-forest area of the country, and compared to the features in Caucasians. Twenty-nine of 51 newly diagnosed cases of AL occurred in childhood, including 19 cases of acute lymphoblastic leukaemia (ALL) and 11 of acute myelogenous leukaemia (AML). The incidence of ALL the AML in Ibadan children was the same, estimated as 0.8 X 10(-5). Thus childhood ALL was about one-third as common in Ibadan as in most developed Caucasian countries. ALL and AML occurred most frequently in the age groups 10-14 and 5-9 years respectively. Six cases of AML were associated with chloromas. Only 2 of the ALL patients survived more than one year after standard chemotherapy. The poor result appeared to be attributable to frequent occurrence among the ALL patients of adverse prognostic factors such as hyperleucocytosis, age less than 2 or greater than 7 years, L2 morphology and low PAS reactivity of the lymphoblasts. Unknown environmental factors are believed to be responsible for the unusual features of AL in children in Ibadan.

The use of lymphocyte transformation and IgM estimation as diagnostic aids in leukaemoid reactions.

Twenty-two patients with lymphocytosis and sometimes accompanied by splenomegaly selected from our difficult diagnostic cases over the past two years are presented. The clinical and laboratory features pointed to one of the following: chronic lymphatic leukaemia without lymphadenopathy, lymphosarcoma or other lymphoreticular tumour, tropical splenomegaly syndrome with a lymphatic leukaemoid reaction. The precise diagnosis was usually made by haemotological laboratory tests - viz. (a) Lymphocytes transformation test (LTT) (b) Serum/Plasma IgM estimation. It was found that: (1) There was markedly raised IgM in the responders i.e. patients with Tropical Splenomegaly Syndrome (TSS) whose spleens regressed following treatment with antimalarials, contrasting the normal levels of IgM in the non-responders to antimalarial therapy. (2) The PHA - Lymphocytes Transformation in the TSS was normal while that of Chronic Lymphatic Leukaemia (CLL) was abnormally low. These combined tests (LTT & IgM) are recommended as investigations for leukaemoid reactions involving lymphocytes.

Hereditary spherocytosis in Nigerians.

Five cases of hereditary spherocytosis in Nigerians are discussed. The clinical presentation and haematological abnormalities are similar to those described in other parts of the world. In one patient family studies were negative. The series does not permit a gene frequency estimate for the country but the cases came from widely separated area, suggesting that no ethnic group has more predilection than others. Splenectomy was uniformly successful.

Interaction of maternal and neonatal cells in mixed-lymphocyte cultures.

One-way and two-way mixed-lymphocyte cultures of cells from male newborn infants and their parents were studied. The tests between the lymphocytes of the parental pairs in the one-way system, in which one population stimulates and the other responds, indicated that maternal lymphocytic response was not generally depressed. However, there was some evidence that the response of the maternal cells was depressed when the cells of the newborn infant were stimulatory. When both maternal and neonatal lymphocytes were viable and the origin of the dividing cells was monitored by using sex chromosomes as markers, it was clearly shown that the division of maternal cells was inhibited, most of the metaphases being XY. The mechanism of this inhibition may depend on contact between viable cells or on the liberation of inhibitory factors by the neonatal cells.