Early responses to chemotherapy detected by pulse cytophotometry.

DNA/cell distributions were recorded by automated cytofluorometry (=pulse cytophotometry) in bone-marrow aspirates of leukaemia and lymphosarcoma patients subjected to chemotherapy. In most cases, early perturbations in DNA/cell histographs were observed, characteristically reflecting the known mode of action of the drugs. These changes in general preceded the clinical observation of drug response. In a series of 23 measurements in 19 patients, a positive correlation between early cytophotometric changes and clinical effects of chemotherapy was observed in 17 patients. Five patients were negative for both cytophotometric and clinical reactions and one patient was probably false-positive. The validity of the assay for early detection of drug resistance in acute leukaemia and related diseases is discussed

A quantitative reverse transcriptase polymerase chain reaction-based assay to detect carcinoma cells in peripheral blood.

The presence of tumour cells in the circulation may predict disease recurrence and metastasis. To improve on existing methods of cytological or immunocytological detection, we have developed a sensitive and quantitative technique for the detection of carcinoma cells in blood, using the reverse transcriptase polymerase chain reaction (RT-PCR) identifying transcripts of the pancarcinoma-associated tumour marker EGP-2 (KSA or 17-1A antigen). The amount of EGP2 mRNA was quantified using an internal recombinant competitor RNA standard with known concentration and which is both reversely transcribed and co-amplified in the same reaction, allowing for a reliable assessment of the initial amount of EGP2 mRNA in the sample. Calibration studies, seeding blood with MCF-7 breast carcinoma cells, showed that the assay can detect ten tumour cells among 1.0 x 10(6) leucocytes. The PCR assay revealed that normal bone marrow expresses low levels of EGP2 mRNA, although immunocytochemistry with the anti-EGP2 MAb MOC31 could not identify any positively stained cell. Analyses using this RT-PCR assay may prove to have applications to the assessment of circulating tumour cells in clinical samples

Bar Diagnostics in Edge-On Spiral Galaxies. III. N-Body Simulations of Disks

Present in over 45% of local spirals, boxy and peanut-shaped bulges are generally interpreted as edge-on bars and may represent a key phase in the evolution of bulges. Aiming to test such claims, the kinematic properties of self-consistent 3D N-body simulations of bar-unstable disks are studied. Using Gauss-Hermite polynomials to describe the stellar kinematics, a number of characteristic bar signatures are identified in edge-on disks: 1) a major-axis light profile with a quasi-exponential central peak and a plateau at moderate radii (Freeman Type II profile); 2) a ``double-hump'' rotation curve; 3) a sometime flat central velocity dispersion peak with a plateau at moderate radii and occasional local central minimum and secondary peak; 4) an h3-V correlation over the projected bar length. All those kinematic features are spatially correlated and can easily be understood from the orbital structure of barred disks. They thus provide a reliable and easy-to-use tool to identify edge-on bars. Interestingly, they are all produced without dissipation and are increasingly realized to be common in spirals, lending support to bar-driven evolution scenarios for bulge formation. So called ``figure-of-eight'' position-velocity diagrams are never observed, as expected for realistic orbital configurations. Although not uniquely related to triaxiality, line-of-sight velocity distributions with a high velocity tail (i.e. an h3-V correlation) appear as particularly promising tracers of bars. The stellar kinematic features identified grow in strength as the bar evolves and vary little for small inclination variations. Many can be used to trace the bar length. Comparisons with observations are encouraging and support the view that boxy and peanut-shaped bulges are simply thick bars viewed edge-on.Comment: 32 pages, 4 figures, AASTeX preprint. Revised following referees' comments. Now accepted for publication in The Astrophysical Journal. We strongly suggest you download the version with full resolution figures at http://www.astro.columbia.edu/~bureau/Publications/Nbody_ApJ04.ps.g

Bar Diagnostics in Edge-On Spiral Galaxies. II. Hydrodynamical Simulations

We develop diagnostics based on gas kinematics to identify the presence of a bar in an edge-on spiral galaxy and determine its orientation. We use position-velocity diagrams (PVDs) obtained by projecting edge-on two-dimensional hydrodynamical simulations of the gas flow in a barred galaxy potential. We show that when a nuclear spiral is formed, the presence of a gap in the PVDs, between the signature of the nuclear spiral and that of the outer parts of the disk, reliably indicates the presence of a bar. This gap is due to the presence of shocks and inflows in the simulations, leading to a depletion of the gas in the outer bar region. If no nuclear spiral signature is present in a PVD, only indirect arguments can be used to argue for the presence of a bar. The shape of the signature of the nuclear spiral, and to a lesser extent that of the outer bar region, allows to determine the orientation of the bar with respect to the line-of-sight. The presence of dust can also help to discriminate between viewing angles on either side of the bar. Simulations covering a large fraction of parameter space constrain the bar properties and mass distribution of observed galaxies. The strongest constraint comes from the presence or absence of the signature of a nuclear spiral in the PVD.Comment: 25 pages (AASTeX, aaspp4.sty), 11 jpg figures. Accepted for publication in The Astrophysical Journal. Online manuscript with PostScript figures available at: http://www.strw.leidenuniv.nl/~bureau/pub_list.htm

Bar Diagnostics in Edge-On Spiral Galaxies. I. The Periodic Orbits Approach

We develop diagnostics to detect the presence and orientation of a bar in an edge-on disk, using its kinematical signature in the position-velocity diagram (PVD) of a spiral galaxy observed edge-on. Using a well-studied barred spiral galaxy mass model, we briefly review the orbital properties of two-dimensional non-axisymmetric disks and identify the main families of periodic orbits. We use those families as building blocks to model real galaxies and calculate the PVDs obtained for various realistic combinations of periodic orbit families and for a number of viewing angles with respect to the bar. We show that the global structure of the PVD is a reliable bar diagnostic in edge-on disks. Specifically, the presence of a gap between the signatures of the families of periodic orbits in the PVD follows directly from the non-homogeneous distribution of the orbits in a barred galaxy. Similarly, material in the two so-called forbidden quadrants of the PVD results from the elongated shape of the orbits. We show how the shape of the signatures of the dominant x1 and x2 families of periodic orbits in the PVD can be used efficiently to determine the viewing angle with respect to the bar and, to a lesser extent, to constrain the mass distribution of an observed galaxy. We also address the limitations of the models when interpreting observational data.Comment: 22 pages, 9 figures (AASTeX, aaspp4.sty). Accepted for publication in The Astrophysical Journa

Identification of Dutch children diagnosed with atopic diseases using prescription data:a validation study

The aim of this study is to validate medication proxies for the identification of children diagnosed with atopic disorders that can be applied in various types of epidemiological research. Records of 7439 children, aged between 0 and 10 years, in the period 2001 until 2010, were retrieved from the Registration Network Groningen database, a general practitioners database in the north-eastern part of the Netherlands. The sensitivity and positive predictive value (PPV) of 22 medication proxies for the identification of children diagnosed with atopic disorders (asthma, atopic dermatitis, and allergic rhinitis) were computed using the registered diagnoses as gold standards. In addition, different capture periods (1 year, half year, and length of study period) for the detection of prescriptions were tested for all the medication proxies. The highest PPV (0.84, 95 % CI 0.81-0.87) in combination with a sufficient sensitivity value (0.54, 95 % CI 0.50-0.57) for the identification of children diagnosed with asthma was yielded for the medication proxy, a parts per thousand yen2 prescriptions for anti-asthma medication within 1 year, including 1 inhaled steroid. PPV and sensitivity were even higher in the age group 6-10 years. The proxies designed for the identification of children diagnosed with atopic dermatitis and allergic rhinitis yielded only high PPVs (a parts per thousand yen0.75) in combination with low sensitivity values (a parts per thousand currency sign0.22). Altering the capture period for the detection of prescriptions to half a year or the length of the study period only affected sensitivity values. Children diagnosed with asthma can be identified reliably with a range of medication proxies. The use of prescription data for the identification of children diagnosed with atopic dermatitis and allergic rhinitis is questionable

Telomere length in breast cancer patients before and after chemotherapy with or without stem cell transplantation

High-dose chemotherapy and peripheral blood stem cell transplantation (PBSCT) may accelerate telomere length loss in haematopoietic stem cells. As data including pre-and post-treatment samples are lacking, we studied leukocyte telomere length and telomerase activity before and after treatment in breast cancer patients randomized to receive 5 adjuvant courses FEC (5-FU, epirubicin and cyclophosphamide) (n= 17), or 4 × FEC followed by high-dose cyclophosphamide, thiotepa, carboplatin and autologous PBSCT (n= 16). Haemoglobin, MCV, leukocyte-and platelet numbers were assessed prior to (t0), 5 months after (t1) and 9 months after chemotherapy (t2); these parameters were decreased at t1 and t2 compared to t0(high-dose: all parameters; standard-dose: leukocytes and platelets), and all parameters were lower after high-dose than standard-dose treatment at t1. Paired individual leukocyte samples of t0 and t1 showed telomere length change (determined by telomere restricted fragment (TRF) assay) ranging from +0.8 to –2.2 kb, with a decreased TRF length in 9 patients of both groups. Telomerase activity (determined by TRAP assay) was below detection limit in leukocyte samples of t0 and t1. Thus, standard-and high-dose chemotherapy negatively affect haematological reconstitution in this setting. In individual patients, telomere length can be remarkably changed following haematological proliferative stress after treatment. © 2001 Cancer Research Campaign www.bjcancer.co

A new in vitro assay for quantitation of chemotherapy-induced mucositis.

Patients receiving high-dose chemotherapy (HD-CT) are at risk of severe mucositis. Most prevention studies evaluate the degree of mucositis on clinical, and therefore subjective, measurements. The aim of this study was to develop an objective in vitro assay of chemotherapy-induced mucositis. Twelve patients with locally advanced breast carcinoma received HD-CT followed by peripheral stem cell reinfusion. Before and twice weekly after HD-CT, the mucosa was evaluated by an oral washing, a buccal smear and the World Health Organization (WHO) toxicity grading; furthermore, blood leucocyte levels were determined. For the oral washings, the percentage of viable epithelial cells was determined by trypan blue dye exclusion and leucocytes were counted by fluorescence microscopy after incubation with acridine orange. Maturity of buccal cells was assessed by staining buccal smears for morphology according to Papanicolaou (Whitacker D and Williams V, 1994). Eight healthy volunteers served as controls. The mean percentage (+/- s.e.m.) of viable oral epithelial cells was stable in controls (44 +/- 2%). In patients, they increased after HD-CT, which was significant after day 7 compared with pretreatment (P < or = 0.05). In addition, a shift from mature to immature epithelial cells in buccal smears was observed. Oral leucocyte levels were closely correlated with the blood leucocyte counts. The WHO score followed the results of these other evaluations with some delay. The viability of buccal cells obtained by oral washings increases after HD-CT. This is possibly because of desquamation of the upper oral mucosa layer, with a shift from mature to more immature cells. These data can be quantitated, and this assay may therefore be useful in studies aimed at prevention of mucositis