European recommendations and quality assurance for cytogenomic analysis of haematological neoplasms.

Cytogenomic investigations of haematological neoplasms, including chromosome banding analysis, fluorescence in situ hybridisation (FISH) and microarray analyses have become increasingly important in the clinical management of patients with haematological neoplasms. The widespread implementation of these techniques in genetic diagnostics has highlighted the need for guidance on the essential criteria to follow when providing cytogenomic testing, regardless of choice of methodology. These recommendations provide an updated, practical and easily available document that will assist laboratories in the choice of testing and methodology enabling them to operate within acceptable standards and maintain a quality service.

Invasive Pasteurella multocida Infections - Report of Five Cases at a Minnesota Hospital, 2014.

During October 2014, the Minnesota Department of Health was notified of five Hospital A patients with Pasteurella multocida bacteraemia; three had died. Human soft tissue infection with P. multocida typically results from cat or dog bites or scratches. Invasive infection, defined as a P. multocida isolate from a usually sterile site, is rare. We evaluated P. multocida isolations at Hospital A, compared with other Minnesota hospitals to understand invasive infection trends. A case was defined as clinically confirmed P. multocida in a Minnesota resident during 2012-2014. All hospital laboratories were queried; Fisher's exact test was used for comparison. Medical charts were reviewed for 2014 Hospital A patients with P. multocida infections. The Minnesota clinical laboratories survey response rate was 79% (63/80). At Hospital A, proportion of P. multocida isolates from usually sterile sites increased from 0% (0/2) during 2012 to 11% (1/9) during 2013, and to 86% (5/6) during 2014. The proportion of patients with P. multocida isolated from sterile sites was 35% (6/17) at Hospital A compared with 10% (58/583) statewide during 2012-2014 combined (P < 0.05). Among 2014 Hospital A patients with invasive P. multocida infection, all five were men; median age was 70 (range: 44-78) years. Four were temporally clustered within a 33-day period; three of those had bacteraemia on admission, making hospital acquisition possible in only one. Among five bacteraemia patients, four had cirrhosis and/or skin ulcerations, and three died. The proportion of invasive P. multocida cases was substantially higher at Hospital A during 2014. No epidemiologic links between patients were found. Three had known pet exposure. Collaborative educational efforts of chronically ill pet owners by physicians and veterinarians can acknowledge the health benefits of pet ownership, while minimizing risk for serious invasive zoonotic infections, including those caused by P. multocida.

Specialist integrated haematological malignancy diagnostic services: an Activity Based Cost (ABC) analysis of a networked laboratory service model.

AIMS: Specialist Integrated Haematological Malignancy Diagnostic Services (SIHMDS) were introduced as a standard of care within the UK National Health Service to reduce diagnostic error and improve clinical outcomes. Two broad models of service delivery have become established: 'co-located' services operating from a single-site and 'networked' services, with geographically separated laboratories linked by common management and information systems. Detailed systematic cost analysis has never been published on any established SIHMDS model. METHODS: We used Activity Based Costing (ABC) to construct a cost model for our regional 'networked' SIHMDS covering a two-million population based on activity in 2011. RESULTS: Overall estimated annual running costs were £1 056 260 per annum (£733 400 excluding consultant costs), with individual running costs for diagnosis, staging, disease monitoring and end of treatment assessment components of £723 138, £55 302, £184 152 and £94 134 per annum, respectively. The cost distribution by department was 28.5% for haematology, 29.5% for histopathology and 42% for genetics laboratories. Costs of the diagnostic pathways varied considerably; pathways for myelodysplastic syndromes and lymphoma were the most expensive and the pathways for essential thrombocythaemia and polycythaemia vera being the least. CONCLUSIONS: ABC analysis enables estimation of running costs of a SIHMDS model comprised of 'networked' laboratories. Similar cost analyses for other SIHMDS models covering varying populations are warranted to optimise quality and cost-effectiveness in delivery of modern haemato-oncology diagnostic services in the UK as well as internationally.

Specific cytogenetic abnormalities are associated with a significantly inferior outcome in children and adolescents with mature B-cell non-Hodgkin's lymphoma: results of the FAB/LMB 96 international study.

Clinical studies showed that advanced stage, high LDH, poor response to reduction therapy and combined bone marrow and central nervous system disease are significantly associated with a decreased event-free survival (EFS) in pediatric mature B-cell non-Hodgkin's lymphoma (B-NHL) treated on FAB/LMB96. Although rearranged MYC/8q24 (R8q24) is characteristic of Burkitt lymphoma (BL), little information is available on other cytogenetic abnormalities and their prognostic importance. We performed an international review of 238 abnormal karyotypes in childhood mature B-NHL treated on FAB/LMB96: 76% BL, 8% Burkitt-like lymphoma, 13% diffuse large B-cell lymphoma (DLBCL). The main BL R8q24-associated chromosomal aberrations were +1q (29%), +7q and del(13q) (14% each). The DLBCL appeared heterogeneous and more complex. Incidence of R8q24 (34%) was higher than reported in adult DLBCL. The prognostic value of cytogenetic abnormalities on EFS was studied by Cox model controlling for the known risk factors: R8q24, +7q and del(13q) were independently associated with a significant inferior EFS (hazard ratio: 6.1 (P=0.030), 2.5 (P=0.015) and 4.0 (P=0.0003), respectively). The adverse prognosis of R8q24 was observed only in DLBCL, whereas del(13q) and +7q had a similar effect in DLBCL and BL. These results emphasize the significant biological heterogeneity and the development of cytogenetic risk-adapted therapy in childhood mature B-NHL.

Genome complexity in acute lymphoblastic leukemia is revealed by array-based comparative genomic hybridization.

Chromosomal abnormalities are important for the classification and risk stratification of patients with acute lymphoblastic leukemia (ALL). However, approximately 30% of childhood and 50% of adult patients lack abnormalities with clinical relevance. Here, we describe the use of array-based comparative genomic hybridization (aCGH) to identify copy number alterations (CNA) in 58 ALL patients. CNA were identified in 83% of cases, and most frequently involved chromosomes 21 (n=42), 9 (n=21), 6 (n=16), 12 (n=11), 15 (n=11), 8 (n=10) and 17 (n=10). Deletions of 6q (del(6q)) were heterogeneous in size, in agreement with previous data, demonstrating the sensitivity of aCGH to measure CNA. Although 9p deletions showed considerable variability in both the extent and location, all encompassed the CDKN2A locus. Six patients showed del(12p), with a common region encompassing the ETV6 gene. Complex CNA were observed involving chromosomes 6 (n=2), 15 (n=2) and 21 (n=11) with multiple regions of loss and gain along each chromosome. Chromosome 21 CNA shared a common region of gain, with associated subtelomeric deletions. Other recurrent findings included dim(13q), dim(16q) and enh(17q). This is the first report of genome-wide detection of CNA in ALL patients using aCGH, and it has demonstrated a higher level of karyotype complexity than anticipated from conventional cytogenetic analysis.

Hybrids of pluripotent and nullipotent human embryonal carcinoma cells: partial retention of a pluripotent phenotype.

To investigate whether the failure of human EC cells that do not differentiate is due to the loss of key differentiation-permissive functions or the acquisition of specific inhibitory functions, we tested the ability to differentiate of 2 hybrids produced between a relatively nullipotent human EC cell line, 2102Ep, and a pluripotent human EC cell line NTERA2. Both hybrids, which exhibited an EC phenotype, were able to differentiate readily in response to retinoic acid. Furthermore, 1 hybrid produced a well-differentiated xenograft tumor, which contained, like the NTERA2 tumors, glandular structures, loose mesenchymal tissues and nodules of cartilage, after injection into a SCID mouse. Thus, the failure of 2102Ep EC cells to differentiate is recessive and due to the loss of a key gene function or functions. Nevertheless, the hybrids differed from the pluripotent NTERA2 line by failing to differentiate in neurons, indicating that 2102Ep cells also had acquired a specific, dominantly-acting, inhibitory mutation specific to the neural lineage. Furthermore, the expression of collagen II by one hybrid before and after induction with retinoic suggested a propensity for spontaneous differentiation not evident in the parental NTERA2 cells. Thus, the mechanisms that restrict the differentiation capacity of the nullipotent 2102Ep line are complex and include both recessive and dominant acting factors.

Blastomycosis. Report of a case with noninvasive, rapid diagnosis of dermal lesions by the Papanicolaou technique.

BACKGROUND: Blastomycosis is a rare fungal infection that occurs most often in young to middle-aged men. A common route of infection is through continued contact with soil, occurring in such occupations as gardening and construction work. The skin and lungs are primarily affected. When the respiratory system is involved, blastomycosis may be misdiagnosed as pulmonary miliary tuberculosis. CASE: A 54-year-old, African American male presented with dry cough and multiple verrucous skin lesions affecting the face, shoulder and legs. Chest roentgenography indicated bilateral lung involvement. The patient died following a short, difficult hospital course. Autopsy revealed verrucous skin lesions with dry, thick, raised margins and central necrosis and umbilication, and severe, diffuse pulmonary involvement clinically and radiologically, resembling miliary tuberculosis. A smear from a leg ulcer stained by the Papanicolaou technique revealed fungi within the giant cells. CONCLUSION: Clinical and radiologic findings alone do not establish the diagnosis of blastomycosis. Respiratory involvement may lead to a misdiagnosis of pulmonary miliary tuberculosis. Sputum and bronchial washing examinations by a laboratory technique are in order. This case indicates that cytologic examination of the exudate from an ulcer may result in a strongly suggestive diagnosis.

Abortion training in family practice residency programs.

BACKGROUND: Despite a relatively fixed demand for services, access to abortion services has become increasingly limited, in part due to fewer training opportunities for OB-GYN residents. The recommended core educational guidelines for family practice residents include voluntary interruption of pregnancy up to 10 weeks gestation, but the availability of and interest in such training have not been reported. METHODS: A survey questionnaire assessing availability of abortion training, percentage of residents participating, program demographics, and resident interest in training was sent to all US family practice residency program directors and randomly selected third-year residents during the 1993-1994 academic year: A total of 301 of 399 (75%) program directors and 253 of 399 (63%) third-year residents completed the questionnaire. RESULTS: Twelve percent of programs nationwide offered abortion training during 1993-1994. Western programs were more likely to offer training (18 of 50 or 36%) than Northeastern (7 of 55 or 12%), Midwestern (6 of 98 or 6%), or Southern programs (4 of 98 or 4%). When available, 45% of trainees chose to participate. Training was primarily in suction curettage up to 12 weeks gestational age. The median duration of training was 4 weeks. Training was provided in both freestanding clinics and hospitals. Religious hospitals were less likely to offer training. CONCLUSIONS: About one of eight family practice residency programs nationwide offer training in abortion. When offered, nearly half of family practice residents choose to participate in this training. Interested family practice residents could be trained to fill an increasing need for physicians willing and able to perform first-trimester abortions.

PIP: In the fall of 1993, 399 family practice residency program directors were asked to complete a questionnaire so researchers could determine the current status of abortion training in family practice residency programs in the US. 301 program directors completed the questionnaire. Only 35 (12%) residency programs provided abortion training to residents. Program directors estimated that 45% of residents from these programs participated in abortion training. Abortion training lasted a median of 4 weeks. Residents performed a median of 10 abortions. They performed them up to 12 weeks gestation (range, 10-20). 30 programs taught suction curettage. Family practice residency programs in the west were more likely to provide this training than the other regions (36% vs. 4% for the south, 6% for the midwest, and 12% for the northeast; p 0.0001). Abortion training takes place primarily in clinics and hospitals (46%, 41%, respectively; 13% for both). Residency programs affiliated with a religious hospital were less likely to offer abortion training than nonaffiliated programs (3.9% vs. 15.7%; p = 0.0335). Religion was the main reason residents refused abortion training. At programs with no abortion training, 46% of senior residents did not know that family practice residents elsewhere received training in first trimester abortions. Only 24% of senior residents who did not know that family practice residents elsewhere received abortion training were interested in receiving abortion training. These findings show that few family practice residents are being trained in performing abortions, particularly in the midwest and the south, which are also the regions with the lowest access to abortion services. North Dakota has only 1 clinic that provides abortions and its physicians come from other states.

Effects of physical stress on complete blood count and venous blood gas profile of individuals with sickle cell trait.

The association between sickle cell trait (SCT) and adverse effects of exercise has been controversial. While individuals with SCT are at higher risk of sudden death, the mechanism for this outcome remains to be elucidated. In order to shed light on this controversy, we have monitored venous blood count and blood gas parameter values in normal and SCT subjects during treadmill exercise. White and red blood cell counts and hemoglobin changed significantly over time in both the SCT and normal groups, with peak exercise values different from pre-exercise or post-exercise values. Red blood cell counts showed significant group-time interaction; increase in count during exercise was accentuated in SCT subjects. All blood gas parameters showed significant changes over time in both groups. O2 content was significantly higher in SCT than AA at all time intervals. O2 saturation, pO2 and CO binding to hemoglobin showed significant group-time interaction. Furthermore, O2 saturation for the combined groups was significantly greater at peak exercise and at rest than before exercise. It is possible that treadmill exercise causes microvascular shunting in SCT subjects, leading to a decrease in the peripheral utilization of oxygen.

Matchmaking in psychotherapy: patient-therapist dimensions and their impact on outcome.

Patient-therapist matching was investigated using recent theoretical and methodological developments to overcome some of the limitations that have hindered similar past efforts. Two hypotheses were tested: (a) Therapeutic outcome is affected by the interaction between patients' and therapists' self-concepts, and (b) increasing anticomplementarity between therapist self-concept and therapists' perceptions of patients' behavior is negatively associated with patient improvement. Data from the first 2 cohorts of the Vanderbilt II Psychotherapy Project were analyzed. They included the therapies of 16 experienced therapists, who saw a total of 48 patients (38 women and 10 men) in time-limited psychotherapy. Patients' ages ranged from 24 to 64 years (M age = 40). Outcome was measured from the perspective of the patient, therapist, and evaluating clinician. Each hypothesis was supported, but not across all 3 outcome perspectives.

Sustained normalization of cardiac function by chelation therapy in thalassaemia major.

The effect of iron chelation using subcutaneous desferrioxamine on the iron-overloaded heart was studied prospectively over 4 years in 23 asymptomatic patients (mean age 13.2 +/- 5.3 years) with thalassaemia major and transfusion-dependent anaemia. The haemoglobin was maintained greater than 10 g/dl by transfusion and chelation therapy to achieve a negative iron balance. Chelation was closely supervised to ensure compliance. Despite an increase in calculated iron load due to transfusion from 34 +/- 27 g to 63 +/- 28 g, there was a sequential fall in serum ferritin levels from 3148 +/- 1956 ng/ml to 2228 +/- 1825 ng/ml (P less than 0.05). Abnormalities of left ventricular (LV) function, assessed by radionuclide angiography, were present at rest or during exercise in 18 of 23 patients (78%) prior to therapy. Normal LV function was restored in 11 of these 18 patients, five continuing to have abnormal function and two dying. There was a significant increase in resting LVEF from 50 +/- 8% to 57 +/- 6% (P less than 0.01). Peak exercise LVEF rose from 51 +/- 11% to 65 +/- 7% (P less than 0.001). We conclude that the common subclinical abnormalities of LV function induced by iron overload in unchelated patients with thalassaemia major can be reversed with long-term subcutaneous infusions of desferrioxamine.

Histoplasmosis: clinical syndromes and management.

The fungus Histoplasma capsulatum produces a spectrum of disease forms ranging from a benign self-limited illness to progressive disseminated disease with a 50 percent mortality rate. The drug of choice, amphotericin B, must be given intravenously over a prolonged course and carries a high incidence of toxicity. Thus, optimal managment of serious forms of histoplasmosis requires considerable clinical judgment.