"Immuno-flowFISH" for the Assessment of Cytogenetic Abnormalities in Chronic Lymphocytic Leukemia.

Imaging flow cytometry is emerging as a diagnostic tool for the assessment of leukemia. It has the functionality of standard flow cytometry and generates high-resolution digital images of each cell with quantifiable numerical data. We demonstrate the use of an automated high-throughput method for performing fluorescence in situ hybridization (FISH) on immunophenotyped whole cells in suspension and analyzed by imaging flow cytometry, a technique called "Immuno-flowFISH". The aim of this study was to demonstrate the application of immuno-flowFISH for the detection of chromosomal abnormalities in CLL, specifically trisomy 12 and del(17p). Mononuclear cells were isolated and immunophenotyped with fluorescently conjugated CD3, CD5, and CD19 monoclonal antibodies. Following fixation, cells were permeabilized, dsDNA denatured and hybridized with chromosome 12 or 17 enumeration (CEP 12 and CEP17) and 17p12 locus-specific FISH probes. Cells were analyzed on the Amnis ImageStream®X Mark II to assess the number and percent FISH-positive CLL cells and the ratio of FISH spot counts for CD5/CD19-positive CLL cells to CD3/CD5-positive T cells (FISH "mean spot ratio"). Deletion of 17p was detected in about 8% of cases to date, with del(17p) ranged from 3.5-22.8% and the FISH "mean spot ratio" 0.86-0.96. Immuno-flowFISH also detected a minimal residual disease case with +12 with a limit of detection of 0.13% and a rare case that presented with atypical phenotype and cytogenetics. Immuno-flowFISH could detect del(17p) in phenotypically identified CD5/CD19-positive B-cells. The 100-fold increase in analyzed cells, as well as the addition of cell phenotype increased the sensitivity and specificity over current clinical FISH testing. Furthermore, immuno-flowFISH analysis demonstrated specific utility in unique clinical scenarios such as residual disease and atypical biology cases which may be of significant benefit with regards to prognostication and MRD analysis. The method will assist in therapeutic decision making and disease monitoring for many hematological malignancies. © 2019 International Society for Advancement of Cytometry.

Non-invasive gastric motility monitor: fast electrogastrogram (fEGG).

We propose a new analysis method to extract the motility information from the electrogastrogram signal that has been recorded at a higher sampler rate than the conventional approaches. This technique utilizes a fourth order Butterworth bandpass filter in extracting the 50-80 cycles per minute (cpm) activity that was previously noted to represent the spike activity range of the cutaneous signals of dogs. Receiver operating characteristics (ROC) analyses have been applied to the processed data to compare the detection performance of our fEGG technique to the conventional approaches that use the slow wave as the reference. The areas under the ROC curves comparing the changes from postprandial stage to fed stage for the fEGG study was found to be 0.961 while for the slow wave it was 0.686. We offer our method as a complementary one to the existing methods.

Consciousness monitoring using near-infrared spectroscopy (NIRS) during high +Gz exposures.

The relationship between human consciousness and oxygen saturation (rSO(2)) in cerebral tissue under high +Gz stress was non-invasively monitored by near-infrared multiple wavelength spectroscopy (NIRS). We studied the drop in rSO(2) levels in human subjects during exposure to various head-to-foot acceleration (+Gz) profiles. These profiles included sustained +Gz plateaus and repeated short duration +Gz pulses of varying duration. The end point in this study was +Gz-induced loss of consciousness (G-LOC). The rSO(2) levels under normal (asymptomatic), almost loss of consciousness (A-LOC) and G-LOC conditions were recorded. Correlations among decrease in rSO(2), +Gz pulse duration, +Gz stress level and incapacitation time (ICAP) after G-LOC were also investigated. It was found that once rSO(2) fell to a certain level, G-LOC occurred. This threshold was repeatable and independent of the +Gz level or duration. It was also observed that the total ICAP after G-LOC was dependent on the length of time that rSO(2) remained below the G-LOC threshold level, i.e. the longer the rSO(2) level remained below the G-LOC induction level, the longer the subject remained unconscious. These results may prove to be useful in designing closed loop control systems for personal protective gear for pilots of high performance aircraft.

[Biological studies and research of immunologic deficiencies in blood donors from a prison environment]. / Explorations biologiques et recherches de déficits immunitaires chez les donneurs de sang en milieu carcéral.

The prison population may be considered as a population at risk for AIDS. Biological parameters were studied in order to detect significant anomalies commonly observed in AIDS patients. With respect to are age matched control population of donors, there are no statistically significant differences concerning the nutritional and inflammatory states of the two populations. The investigation of the humoral immunity shows comparable levels of circulating antibodies in the two groups: a high level of anti-cytomegalovirus and anti-herpes antibodies is more frequently found in the penal population. The markers for hepatitis B were also studied. None of the individuals is a carrier of the HBs antigen. The percentage of individuals having biological markers of hepatitis B is higher in the at risk group (45%) than in the control group (10%). The evaluation of the cell-mediated immunity shows that there are no significant differences between the mean values found in the two groups for OKT3, T11, OKT4 and OKT8. There is no inversed OKT4/OKT8 ratio in the at risk group while one donor in the control group shows an inversed OKT4/OKT8 ratio.