Cytogenetic basis of acute myeloid leukemia.

The chromosomes of 12 adult patients with acute leukemia were analyzed by conventional means and by Giemsa and centromeric banding techniques. Acute myeloblastic leukemia was diagnosed in 7, acute myelomonocytic leukemia in 2, and acute undifferentiated leukemia in 3. Bone marrow was aspirated from patients when in relapse or remission, and both euploid and aneuploid cells were examined. All patients showed trisomy no. 9 and many showed additional numerical or structural changes in some or all their cells. These changes included monosomy no. 21 and/or monosomy no. 8. The proportion of trisomy no. 9 cells was 30-50% in patients in full remission and up to 100% in patients in relapse; thus trisomy no. 9 might be an important marker of leukemic cells. A mechanism was proposed to explain the induction and selection of the trisomy no. 9 karotype.

Electric field-mediated gene transfer into K562 cells: optimization of parameters affecting efficiency.

Since hemopoietic cells are refractory to transfection by conventional chemical means, we have developed a reliable and efficient gene transfer system for K562 cells which uses electric field-mediated gene transfer (EFMGT). EFMGT involves the exposure of cells in suspension to an electric field which transiently allows the entry of DNA into the cell and its subsequent integration and expression. Plasmids bearing the neo gene were used to identify and select transfected clonogenic cells manifested by geneticin resistance in semisolid medium. Transfection efficiency is significantly affected by the following variables: voltage, capacitance, time constant, number of pulses, buffer type and temperature, DNA concentration, configuration, and promoter type. Cell cycle status also appears to be critical as shown in studies employing aphidicolin synchronization. Using optimal conditions, we have consistently achieved a transfection efficiency of 0.3-0.4% of clonogenic cells per microgram DNA. Stability of neo gene expression was also demonstrated after 4 months in nonselective culture conditions. This level of efficiency compares favorably with other reports of gene transfer into human hemopoietic progenitor cells.

Clinician referrals for stress echocardiography: are we compliant with the NICE guidelines?

Accurate diagnosis of stable angina is of paramount importance, and where possible, this should be based on clinical history. In cases of uncertainty, the National Institute for Health and Care Excellence (NICE) provides a framework for assisting diagnosis based on pre-test likelihood (PTL) of coronary artery disease. Functional testing such as stress echocardiography (SE) is recommended as a first-line investigation in patients with PTL of 30-60%. This study evaluated hospital clinicians' adherence to this recommendation. A prospective analysis of patients referred for SE at a district general hospital between March and May 2013 was performed. Data were extracted from an electronic database of SE reports and medical notes. A total of 193 patients were assessed. The most common PTL was 61-90%, accounting for 40% of the cohort. Of them, 14% had a PTL of 30-60%. Of these, 15% had positive SE; 57% described non-anginal pain, as defined by NICE, of whom only nine cases had SE positivity. None of these patients required revascularisation. Findings suggest that SE is being used in a much broader selection group than advocated by NICE. This may often be for its exclusion value rather than to stratify risk. Although utility may be justified in high-risk patients to avoid proceeding directly to invasive angiography, SE appears to add little in those with non-anginal pain and with low PTL. Greater focus should be directed towards characterisation of symptoms, which may negate the need for subsequent investigation.