Cytogenetic culture failure and its causes in hematological disorders; a single centre perspective.

OBJECTIVE: To highlight the reasons of culture failure in bone marrow aspirate samples sent for Cytogenetic analysis and to identify the associated parameters causing this impact. METHODOLOGY: This is a retrospective cross-sectional study conducted in the Clinical and Molecular Cytogenetics Laboratory of NIBD Hospital, Karachi, Pakistan. The rates of culture failure are assessed from the year 2017-2020 along with their reasons. Bone Marrow aspirate samples of patients with hematological malignancies were cultured for chromosomal analysis, both at the time of diagnosis or relapse. Statistical analysis was performed using SPSS version 25. RESULTS: A total of 1061 bone marrow aspirate samples were assessed for cytogenetic culture failures from the duration of 2017 to 2020. Ratio of males was predominantly higher i.e. 62.7% than female 37.3% with Mean ± SD age was 36.78 ± 18.94. Frequency of culture failure in the year 2020 was relatively high 20% as compared to the preceding years i.e. 8% in 2017, 6% in 2018, 7% in 2019. However, the patients were diagnosed with the following hematological malignancies; ALL 23%, CML 17.1%, AML 16.5% and AA 12.5%. Among the reasons of culture failure, cytogenetic analysis of patients with on-going chemo resulted in significant culture failures with p-value < 0.001 and the hematological malignancy, Acute Promyelocytic Leukemia, significantly impacted the growth of bone marrow aspirate cultures, with p-value < 0.001. CONCLUSION: Significant findings were associated with causative factors of culture failure including on-going treatment and sample issues of clotted bone marrow as well as with the clinical diagnosis. These evaluations facilitated in overcoming the rise in culture failures. As per our knowledge, no such data, discussing the effects of various parameters such as sample quality, diagnosis, effects of treatment etc., has been documented previously.

A Familial Case of Robertsonian Translocation 13;14: Case Report.

Robertsonian translocations are the most common form of chromosomal abnormalities that specifically involve the acrocentric chromosomes. Robertsonian translocation between chromosomes 13,14 and 14,21 are the most frequently reported. Infertility is common in genetically balanced carriers of these translocations, and their conceptions are more likely to have imbalances. Here we have reported a case of an 18-year-old female presenting with a complaint of primary amenorrhea. Cytogenetic analysis revealed a familial case of maternally inherited Robertsonian translocation (rob(13;14)(q10;q10)) affecting all the siblings. Genetic counseling and genetic testing are recommended especially in familial cases as the carriers are normal but can lead to several genetic disorders in their future generation.

Exogenous salicylic acid-induced drought stress tolerance in wheat (Triticum aestivum L.) grown under hydroponic culture.

Wheat is an important cereal crop, which is adversely affected by water deficit stress. The effect of induced stress can be reduced by the application of salicylic acid (SA). With the objective to combat drought stress in wheat, an experiment was conducted in greenhouse under hydroponic conditions. The treatments consisted of (a) no drought (DD0 = 0 MPa), mild drought (DD1 = -0.40 MPa) and severe drought (DD2 = -0.60 MPa) by applying PEG-8000, (b) two contrasting wheat varieties Barani-17 (drought tolerant) and Anaj-17 (drought-sensitive), and (c) foliar treatments of salicylic acid (0, 50 mM, 75 mM, and 100 mM). Evaluation of wheat plants regarding biochemical, physiological, and morphological attributes were rendered after harvesting of plants. Statistically, maximum shoot and root fresh and dry weights (18.77, 11.15 and 1.99, 1.81 g, respectively) were recorded in cultivar Barani-17 under no drought condition with the application of SA (100 mM). While, minimum shoot and root fresh and dry weights (6.65, 3.14 and 0.73, 0.61 g, respectively) were recorded in cultivar Anaj-2017 under mild drought stress without SA application. The maximum shoot length (68.0 cm) was observed in cultivar Barani-2017 under no drought condition with the application of SA (100 mM). While, maximum root length (59.67 cm) was recorded in cultivar Anaj-17 under moderate drought stress without application of SA. Further, minimum shoot length (28.67 cm) was recorded in Anaj-17 under moderate drought stress without SA application. Minimum root length (38.67 cm) was recorded in cultivar Barani-17 under no drought condition without SA application. Furthermore, maximum physio-biochemical traits, including membrane stability index (MSI), chlorophyl content, photosynthetic rates, stomatal conductance, antioxidant enzymatic activities and relative water content (RWC) were found highest in cultivar Barani-17 under no drought stress and SA application at 100 mM. However, minimum values of these traits were recorded in cultivar Anaj-17 under severe drought stress without SA application. Our results also demonstrated that under severe drought, application of SA at 100 mM significantly increased leaf nitrogen (N), phosphrus (P) and potassium (K) contents and cultivar Barani-17 demonstrated significantly higher values than Anaj-17. The obtained results also indicated that the cultivation of wheat under drought stress conditions noticeably declines the morphological, physiological, and biochemical attributes of the plants. However, the exogenous application of SA had a positive impact on wheat crop for enhancing its productivity.

Complex Karyotype in Hematological Diseases: A 6-Year Single Centre Study from Pakistan.

BACKGROUND: Most of the hematological disorders are heterogenous with regard to morphology, immunophenotype, and genetic rearrangements. Multiple recurrent chromosomal aberrations have been identified by conventional cytogenetic analysis, which is now widely recognized as one of the most important diagnostic and prognostic determinants in these patients. Though rarer, complex karyotype has been associated with worst prognosis. MATERIALS AND METHODS: A total of 1185 bone marrow or peripheral blood cytogenetics samples were taken with different hematological diseases. They included both benign and malignant disease entities. In each case, cells were cultured and conventional cytogenetic analysis was performed. RESULTS: Among 1185 subjects, 41 (3.4%) patients possessed complex cytogenetic abnormalities. Out of these 41, 33 (80%) were males. The mean age was 37 years (median age 39 years). Myelodysplastic syndromes had the most numbers of complex karyotypes (8%), followed by acute myeloid leukemia (7%) and acute lymphoblastic leukemia (4%). Also we found few patients with acute promyelocytic leukemia, aplastic anemia , chronic myeloid leukemia, and diffuse large B cell Lymphoma possessing complex karyotype. Frequencies of different cytogenetic abnormalities were assessed with respect to disease as well as independently. Trisomy 21 was the most common chromosomal abnormality found in 28% of patients. CONCLUSION: Complex karyotype was most frequently associated with myelodysplastic syndromes and acute myeloid leukemia. Trisomy 21 and deletion 5q were the commonest cytogenetic abnormalities found. We also assessed complex karyotype in benign diseases and detected one patient of aplastic anemia with complex karyotype. This is the first study highlighting the presence of complex karyotypes in hematological disorders in our region.