Fifteen cases of t(1;19)(q23;p13.3) identified in an Australian series of 122 children and 80 adults with acute lymphoblastic leukemia

The t(1;19)(q23;p13) has been reported in up to 6% of cytogenetically abnormal cases of acute lymphoblastic leukaemia (ALL), associated with a pre-B-ALL phenotype. In the 5-year period 1995-1999, we detected t(1;19) in 13 children and 2 adults with newly diagnosed ALL. This represented 10% of pediatric and 2.5% of adult diagnostic ALL samples successfully cultured in one center during this time. There were 9 males and 6 females. The mean age at diagnosis for the 13 children was 6.5 years (range 1.5 to 14 years) and the 2 adults were aged 42 and 45 years. The unbalanced t(1;19) occurred in 7 of 13 children (54%), contrary to the reported excess of unbalanced translocations at 75%; both adults had the unbalanced translocation. At diagnosis, the t(1;19) was the sole abnormality in 4 patients (26%), and in the remainder (74%) was part of a complex karyotype, which included i(7q) (2 patients), hyperdiploidy (2 patients) and del(6q) (2 patients). Correlation of karyotype with white cell, blast and platelet counts, cell surface markers, initial response to chemotherapy and short-term outcome showed no difference between the balanced and unbalanced forms of the translocation in children or whether t(1;19) was present as the sole abnormality or part of a complex karyotype.     

Blood pressure reactions to acute psychological stress and future blood pressure status: a 10-year follow-up of men in the Whitehall II study

OBJECTIVE: The aim of this study was to examine whether blood pressure reactions to mental stress predicted future blood pressure and hypertension. METHODS: Blood pressure was recorded at an initial medical screening examination after which blood pressure reactions to a mental stress task were determined. A follow-up screening assessment of blood pressure and antihypertensive medication status was undertaken 10 years later. Data were available for 796 male public servants, between 35 and 55 years of age upon entry to the study. RESULTS: Systolic blood pressure reactions to mental stress were positively correlated with follow-up screening systolic blood pressure and to a lesser extent, follow-up diastolic pressure. In multivariate tests, by far the strongest predictors of follow-up blood pressures were initial screening blood pressures. In the case of follow-up systolic blood pressure, systolic reactions to stress emerged as an additional predictor of follow-up systolic blood pressure. With regard to follow-up diastolic blood pressure, reactivity did not enter the analogous equations. The same outcomes emerged when the analyses were adjusted for medication status. When hypertension at 10-year follow-up was the focus, both systolic and diastolic reactions to stress were predictive. However, with correction for age and initial screening blood pressure, these associations were no longer statistically significant. CONCLUSIONS: The results of this study provide modest support for the hypothesis that heightened blood pressure reactions to mental stress contribute to the development of high blood pressure. At the same time, they question the clinical utility of stress testing as a prognostic device.     

A rapid and reproducible method for the analysis of immune complexes using affinity chromatography and Western blotting

A new procedure which couples different analytical techniques in a format permitting the rapid analysis of immune complex components is described. Complexes obtained from sera by polyethylene glycol (PEG) precipitation were resuspended and then added, using a batch method, to antibody coupled to Sepharose beads. Antibody directed against either human C1q or human C3c were used in the present study. Bound immune complexes were washed and then eluted from the Sepharose by sodium dodecyl sulphate (SDS) treatment and simultaneously reduced with dithiothreitol. Individual components were separated by SDS gradient polyacrylamide gel electrophoresis and then transferred to nitrocellulose by Western blotting. Individual strips of nitrocellulose were investigated using specific antisera and a radiolabelled probe. Immune complexes (IC) isolated from the sera of 7 rheumatoid arthritis (RA) patients were analysed using this method and the results obtained for both affinity adsorbents compared.     

A microcomputer-based data acquisition system for continuous recording of feeding and drinking by rats

A monitoring system to continuously record the daily pattern of drinking and eating of rats is described. This system, based on a North Star microcomputer, can record the amount of food ingested with a temporal resolution of +/- 1.0 second and quantitative accuracy within +/- 5%. Drinking behavior is detected using a drinkometer which also has a temporal resolution of +/- 1.0 second. Data are analyzed by computer to determine absolute amounts of consumption and patterns of intake. The patterns of feeding and drinking recorded by this system are similar to those observed using other monitoring devices.     

Rat self administration of ethanol: Enhancement by darkness and exogenous melatonin

Using an FT 60 schedule, rats on 100% free feeding tested in the dark phase of a 12:12 light-dark cycle were trained to self-administer ethanol intravenously. The effect was dose-dependent with 20% ethanol being the preferred dose as measured by the number of infusions. Daily administration of 1.5 mg/kg melatonin significantly increased ethanol self-injection in the dark but not in the light. The time of day of testing and/or drug administration may be an important variable in studies on self-administration of drugs. Testing in the dark may eliminate the need for reducing body weight when inducing self-administration of ethanol.     

A novel ELISpot method for adherent cells

The purpose of this study was to develop an enzyme-linked immunospot assay (ELISpot assay) that can be used with human adherent cells. While standard enzyme-linked immunosorbent assays (ELISAs) are available and widely used and ELISpot assays are used for nonadherent lymphocytes, no ELISpot assay has been developed for adherent cells. We used primary human fibroblasts from four different tissues (myometrium, lung, gingiva, and orbit), either unstimulated or interleukin (IL)-1beta-activated, to evaluate an ELISpot assay. Antibody pairs for IL-6 and IL-8 were used and results were compared to a standard ELISA. We found that we could reliably detect IL-6 and IL-8 spots with as few as 10 fibroblasts. Optimal cell numbers were 50 cells per well incubated for 8 h, although spots appeared as early as 2 h after incubation. Spots were absent when cells, primary, or secondary anti-cytokine antibodies were omitted from the protocol. Spot number and size can be ascertained using current automated ELISpot reader technology. The frequency of IL-6 and IL-8-producing human fibroblasts could also be determined. For example, 60% of the lung fibroblasts express IL-6, but IL-8 can be detected from only 40% of the cells. Approximately 80% of the human orbital fibroblasts make IL-6, whereas approximately 50% generate IL-8 following IL-1beta stimulation. These new findings show that fibroblasts from different human tissues display different frequencies of cytokine production and this further supports the concept of fibroblast diversity. The sensitivity of this new ELISpot assay is adequate for cytokine detection in just a few cells, unlike the standard ELISA. It should permit ascertaining the frequency of fibroblasts and other adherent cells that produce cytokines and, if desired, can be used in tandem with a standard ELISA to determine total cytokine produced. Moreover, the assay is suitable for normal human adherent cells that are often short-lived and difficult to cultivate.     

Abnormal expression of the cell cycle regulators P16 and CDK4 in Alzheimer's disease.

In this study, we demonstrate that two important regulators of the cell cycle, cyclin-dependent kinase-4 and its inhibitor p16, are increased in the brains of cases of Alzheimer's disease patients compared with age-matched controls. Both proteins are increased in the pyramidal neurons of the hippocampus, including those neurons containing neurofibrillary tangles and granulovacuolar degeneration. As p16 is not normally found in terminally differentiated neurons, it seems paradoxical that it is increased in Alzheimer's disease unless it is responding to increases in cyclin-dependent kinase-4 or other cell cycle regulators. Induction of the latter, a protein that signals re-entry and progression through the cell cycle, may itself be the consequence of alpha response to a growth stimulus. Re-entry into the cell cycle is likely deleterious in terminally differentiated neurons and may contribute to the biochemical abnormalities, such as oxidative stress and hyperphosphorylated tau protein, as well as the neuronal degeneration characteristic of the pathology of Alzheimer's disease.     

Signaling mechanisms regulating self-renewal and differentiation of pluripotent embryonic stem cells

An ability to propagate pluripotent embryonic cells in culture is the foundation both for defined germline modification in experimental rodents and for future possibilities for broad-based cellular transplantation therapies in humans. Yet, the molecular basis of the self-renewing pluripotent phenotype remains ill-defined. The relationship between factors that influence embryonic stem cell propagation in vitro and mechanisms of stem cell regulation operative in the embryo is also uncertain. In this article we discuss the role of intracellular signalling pathways in the maintenance of pluripotency and induction of differentiation in embryonic stem cell cultures and the mammalian embryo.