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PubMed1988

Microcomputer software and interface for control of a microscope scanning stage.

Motorized scanning stages are valuable in microscopy systems that employ digital image analysis and for the development of semiautomatic computer-assisted microscope systems; the development of standard software "tools" to control such stages will facilitate their integration into a variety of computer-based systems. A set of Microsoft BASIC and Turbo PASCAL programs that interfaces a microprocessor-controlled stepper motor microscope stage (MDACE 1000) to an IBM PC or PC-AT or compatible microcomputer via a serial interface (RS-232) is described. These programs can be integrated into other software written in either BASIC or PASCAL, or used via a menu program that directs the routines to control scanning patterns and to locate the microscope stage to a selected area of the slide. Coordinates of significant events on a slide can be stored on a disk file to allow future examination. The software and interface also provide control of a filter wheel in the microscope for use in multicolor fluorescence assays.

Computers↗
PubMed1987

Appearance of polychromic endothelial pigment.

Multicolored corneal endothelial pigment was noted in patients on routine specular microscopy, confirming the presence of chromatic pigment deposits at the corneal endothelium. These deposits were polychromic, exhibiting blue, green, red, pink, orange, gold, and yellow colors. These pigment deposits were of different sizes and shapes and often seemed to surround darkened cells or clusters of cells. The variations in color may be the result of differing concentrations of melanin in iris or other pigmented cells that adhere to the posterior surface of the corneal endothelium. To our knowledge, this is the first report of the observation of polychromic endothelial pigment. We encourage the use of color film for specular microscopy to help detect and identify this pigment further.

Aged↗
PubMed1985

[Malarial problems and the trans-Saharan highway].

In Maghreb, malaria eradication is near. In particular, no indigenous strains of P. falciparum have been found for ten years. The few remaining cases of malaria are of P. vivax. In the Sahelian area, on the other hand, malaria eradication failed with P. falciparum and the efficient vector A. gambiae (s.l.). Until now, the Sahara desert separated these two areas. With construction of a trans-saharan highway, the situation is very different. Now, tropical malaria circulates in Maghreb and Sahara oases in the same way as vectors. However, it seems that local vectors in Maghreb (A. labranchiae) and in Sahara (A. multicolor et A. sergenti) are not efficient vectors for tropical strain of P. falciparum. These observations must be confirmed because there are probably different anopheles populations with different vectorial capacity in the same anopheles species and different plasmodial strains with different infective capacity in the same plasmodial species. Meanwhile a dangerous potential situation exists, the temporary or permanent colonization of some stations in Sahara, in Maghreb (?) by A. gambiae the efficient vector of tropical strains of P. falciparum. From now on, important efforts of Prevention and Control for vectors and malaria must be developed with large intensity in Saharan oases and in Maghreb.

Africa, Northern↗
PubMed1979

[Histologic study of the teleost liver. II. The blood vessel system].

In the liver of the teleosts investigated in the present study the sinusoidal region shows a space of DISSE, which contains numerous microvilli originating from the hepatocytes. In some species, especially in Tetraodon leiurus, there are also bundles of collagenous fibrils. In the DISSE's space of larger sinusoids (transition sinusoids) sections of filament-rich cells are found. These are sometimes interconnected by desmosomes and can be interpreted as processes of smooth muscle cells from the region of the venae hepaticae. The endothelium of smaller sinusoids is fenestrated and shows micropinocytotic activity. The endothelia of the transition sinusoids and of the venae hepaticae are endowed with structures, which can be interpreted as macrovesicles. In the sinusoidal region true KUPFFER-cells and ITO-cells could not be observed. Nevertheless, the close location of granulocytes to the sinusoidal endothelium suggests that phagocytotic processes cannot be excluded for the sinusoidal region. Exceptionally, in Hemihaplochromis multicolor there were also signs of possible phagocytosis by sinusoidal endothelial cells. The chemomorphology of the sinusoidal region, above all the evidence of alkaline phosphatase, shows great differences between species. The examination of the larger blood-vessels in the liver of Haplochromis burtoni reveals venae portae with very thin walls and venae hepaticae with thick walls, which contain smooth muscle cells. Granulocytes and melanocytes are abundant in the wall of the venae hepaticae. This phenomenon indicates that the defence-functions, which in the liver of higher vertebrates are carried out by the sinusoids (KUPFFER-cells), are undertaken by the region of the venae hepaticae in the liver of Haplochromis burtoni, which is free of KUPFFER-cells. On their extrahepatic course the venae portae are surrounded by a sleeve of exocrine pancreatic tissue, which accompanies the vessels deep into the liver. The pancreatic cells bordering the thin-walled venae portae have sparse microvilli indicating a transfer of substances between venous blood and pancreas similar to the sinusoidal region of the liver. Furthermore, the investigation resulted in a clue to the innervation of the exocrine pancreas.

Animals↗
PubMed1984

Human lymphocyte subpopulations identified by using three-color immunofluorescence and flow cytometry analysis: correlation of Leu-2, Leu-3, Leu-7, Leu-8, and Leu-11 cell surface antigen expression.

Three-color immunofluorescence has been used to determine the co-expression of cell surface antigens on human peripheral blood lymphocytes. Monoclonal antibodies or avidin were coupled to either FITC (green), phycoerythrin (orange), or Texas Red (red) fluorochromes. These three fluorochromes could be independently measured by using a dual laser FACS IV system equipped with an argon ion laser (488 nm) and a dye laser (600 nm). Human peripheral blood lymphocytes were stained with the following combinations of reagents: (1) FITC anti-Leu-11a + PE anti-Leu-2a + TR avidin/biotin anti-Leu-7; (2) FITC anti-Leu-11a + PE anti-Leu-3a + TR avidin/biotin anti-Leu-7; (3) FITC anti-Leu-8 + PE anti-Leu-2a + TR avidin/biotin anti-Leu-7; and (4) FITC anti-Leu-11a + PE anti-Leu-2 + TR avidin/biotin anti-Leu-8. The light scatter, green fluorescence, orange fluorescence, and red fluorescence signals for each sample were stored by a Consort 40 PDP/11 computer in list mode files. Sequential reanalysis of the data directly demonstrated the existence of several unrecognized subpopulations of lymphocytes. Previously, we reported that the anti-Leu-7 and anti-Leu-11 antibodies can be used to identify discrete subsets of human NK cells with distinct functional capacities. In this report, we show that these subsets can be further subdivided on the basis of Leu-8 and Leu-2 expression. Thus, these studies illustrate how multicolor and multiparameter flow cytometry can further our understanding of cellular heterogeneity within this group of lymphocytes.

Adult↗
PubMed1983

Relationship between cell size and weight of the human liver. An automated morphometric study.

Using a machine-vision system for the analysis of multicolor multicellular images, an algorithm, consisting of cellular logic operators, was developed for estimating the average cross-sectional area of cells in the liver. A total of 28 three-color histologic images from 22 variable-weight autopsied livers were digitized and processed. The data collected showed a positive correlation, albeit weak, between the weight of the liver and the cell size. This correlation was not visually evident, which demonstrates the utility of machine vision in histopathology.

Adult↗
PubMed1978

Olfactory neuroepithelioma in a domestic carp (Cyprinus carpio).

A well-differentiated olfactory neuroepithelioma was found protruding from the left olfactory fossa of a 6-year-old domestic multicolored carp, Cyprinus carpio. Following diagnostic biopsy the neoplasm continued to grow. At necropsy, the tumor was found to extend beyond the olfactory chamber into the intracranial portion of the olfactory bulb, but no distant metastases could be detected. Histologically, the neoplasm had a remarkably organoid appearance, being composed of imperfect reproductions of the normal olfactory organ including olfactory membrane, axonal bundles, and glial tissue. Electron microscopy demonstrated the presence of numerous ciliated olfactory sensory cells as well as sustentacular, mucous, and basal cells within the neoplastic simulations of olfactory membrane. These observations, together with those from two previous case reports of olfactory neuroepitheliomas in teleosts, suggest that this type of tumor tends to be more highly differentiated than are neoplasms of similar origin in mammals.

Animals↗
PubMed1980

Long-term efficacy and safety of benoxaprofen: comparison with aspirin and ibuprofen in patients with active rheumatoid arthritis.

Benoxaprofen, 400 to 600 mg daily, was compared to aspirin, 4,000 to 6,000 mg daily, or ibuprofen, 1,600 to 2,400 mg daily, in 2 multicolor clinical trials. The study design was double-blind and provided 28 wk of active drug therapy. Statistical analysis of the results showed benoxaprofen to be at least as effective as the 2 control drugs. More patients taking ibuprofen or aspirin discontinued therapy than patients taking benoxaprofen. Side effects occurred more frequently and lasted longer in patients who took aspirin during the study. Clinical laboratory examinations supported the long-term safety of benoxaprofen.

Adult↗
PubMed1982

Fiber optics in medicine.

The most significant and prevalent applications of fiber optics in medicine are in the imaging and illumination components of endoscopes. Flexible and rigid multifibers composed of step-index fibers and graded-index imaging rods are extensively used for visualization of internal organs and tissue which are accessible through natural openings or transcutaneously. This article reviews the evolution of the fiber-optic endoscope over the past 20 years. The fabrication techniques of optical fibers for imaging and illumination are considered in juxtaposition to their current applications in communications with emphasis on the different technologies involved. The design of distinct endoscopes is described with a detailed review of their use in various medical specialities. Low-loss optical fibers are employed to transmit laser energy for surgery and photocoagulation. Multicolor laser light is transmitted through a single thin optical fiber to provide adequate illumination for viewing and color photography. Past attempts to develop plastic imaging multifibers and their future potential viewing and color photography. Past attempts to develop plastic imaging multifibers and their future potential are discussed. Other uses of fiber optics in medicine include remote spectrophotometry, pressure and position sensing, or scintillation counting. These and other applications, such as intravascular pressure transducers and in vivo oximeters, are described.

Bronchoscopes↗
PubMedDate not supplied

[Periodical structure of the constant electrical field in a higher plant].

The constant electrical field of a higher plant Tradescancia multicolor was studied. It has been shown that the constant electrical field is a quasiperiodical structure correlating with a physiological structure of the plant. The node and internode differ from one another by the largest difference of the electrical potential, the plant node rather than the internode having a more positive difference with respect to the plant root.

Electric Conductivity↗
PubMedDate not supplied

[Electrical nature of the spread of the variable potential of Tradescantia].

Propagation of action potential in Tradescantia multicolor stem induced by the damage of one of the leaves with a burn was studied by extracellular recording. The dependence of signal propagation velocity on the stem electric resistance was obtained. The effect of plant physiological state on signal propagation velocity was shown. The data obtained are explained by the presence of vector properties in the conducting systems stipulated by the gradients of its electrical properties.

Electric Conductivity↗
PubMed1980

Differentiation of cells in abnormal human liver tissue by computer image processing: a preliminary investigation to diagnostic microscopy.

The initial development of algorithms for the analysis of multicolor multicellular images of liver tissue sections is described. Using a three-color image recorded by an automated light microscope, a sequence of cellular logic operators was applied to the task of differentiating between hepatocyte and leukocyte nuclei in a section taking advantage of both spatial and chromatic features, namely, that hepatocyte nuclei usually appear larger than those of leukocytes and, in red illumination, are likely to be more transparent. Optimally, the developed algorithms properly differentiated 90% of the hepatocyte nuclie and 85% of the leukocyte nuclie; this performance level appears to equal that of a microscopist. The microscopist, however, rarely analyses all cells; thus, computer differentiation of cells eventually may provide an important new dimension to clinical histopathology.

Cell Nucleus↗
PubMed1994

A method to stain nuclei of Drosophila for confocal microscopy.

We report a method of staining nurse cell and follicle cell nuclei in Drosophila ovaries and nuclei in Drosophila embryos with the fluorescent dye propidium iodide. This technique was used to replace more commonly used 4', 6-diamidino-2-phenylindole (DAPI) and Hoechst staining as a method of visualizing nuclear material in Drosophila. Propidium iodide has its maximum excitation at about 530 nm and maximum fluorescence at 615 nm, and therefore it can be used as a fluorescent marker with confocal microscopes that do not have a UV excitation source. Another advantage of the described method is the convenience of simultaneous use of fluorescein as a second fluorophore in multicolor fluorescence. We show that the nuclear material in Drosophila ovaries and early embryos can be visualized with propidium iodide using both confocal and conventional fluorescence microscopy. We also test the combination of two fluorophores-propidium iodide for nuclear staining and fluorescein-labeled phalloidin for membrane-bound actin--in the same tissue.

Animals↗
PubMed1994

HLA-DR-, CD33+, CD56+, CD16- myeloid/natural killer cell acute leukemia: a previously unrecognized form of acute leukemia potentially misdiagnosed as French-American-British acute myeloid leukemia-M3.

We have identified and characterized a previously unrecognized form of acute leukemia that shares features of both myeloid and natural killer (NK) cells. From a consecutive series of 350 cases of adult de novo acute myeloid leukemia (AML), we identified 20 cases (6%) with a unique immunophenotype: CD33+, CD56+, CD11a+, CD13lo, CD15lo, CD34+/-, HLA-DR-, CD16-. Multicolor flow cytometric assays confirmed the coexpression of myeloid (CD33, CD13, CD15) and NK cell-associated (CD56) antigens in each case, whereas reverse transcription polymerase chain reaction (RT-PCR) assays confirmed the identity of CD56 (neural cell adhesion molecule) in leukemic blasts. Although two cases expressed CD4, no case expressed CD2, CD3, or CD8 and no case showed clonal rearrangement of genes encoding the T-cell receptor (TCR beta, gamma, delta). Leukemic blasts in the majority of cases shared unique morphologic features (deeply invaginated nuclear membranes, scant cytoplasm with fine azurophilic granularity, and finely granular Sudan black B and myeloperoxidase cytochemical reactivity) that were remarkably similar to those of acute promyelocytic leukemia (APL); particularly the microgranular variant (FAB AML-M3v). However, all 20 cases lacked the t(15;17) and 17 cases tested lacked the promyelocytic/retinoic acid receptor alpha (RAR alpha) fusion transcript in RT-PCR assays; 12 cases had 46,XX or 46,XY karyotypes, whereas 2 cases had abnormalities of chromosome 17q: 1 with del(17)(q25) and the other with t(11;17)(q23;q21) and the promyelocytic leukemia zinc finger/RAR alpha fusion transcript. All cases tested (6/20), including the case with t(11;17), failed to differentiate in vitro in response to all-trans retinoic acid (ATRA), suggesting that these cases may account for some APLs that have not shown a clinical response to ATRA. Four of 6 cases tested showed functional NK cell-mediated cytotoxicity, suggesting a relationship between these unique CD33+, CD56+, CD16- acute leukemias and normal CD56+, CD16- NK precursor cells. Using a combination of panning and multiparameter flow cytometric sorting, we identified a normal CD56+, CD33+, CD16- counterpart cell at a frequency of 1% to 2% in the peripheral blood of healthy individuals. Our studies suggest that this form of acute leukemia may arise from transformation of a precursor cell common to both the myeloid and NK cell lineages; thus we propose the designation myeloid/NK acute leukemia. Recognition of this new leukemic entity will be important in distinguishing these ATRA-nonresponsive cases from ATRA-responsive true APL.

Acute Disease↗
PubMed1994

Harvesting, characterization, and culture of CD34+ cells from human bone marrow, peripheral blood, and cord blood.

Stem and progenitor cells from a variety of sources including bone marrow, cord blood, and peripheral blood have been used for transplantation. This study compares CD34 cells from all three sources. Flow cytometry analysis of CD34 cells in multiple samples of normal peripheral blood and patient peripheral blood mobilized with chemotherapy (cyclophosphamide/VP16), chemotherapy plus granulocyte colony stimulating factor (G-CSF), and G-CSF alone were compared to bone marrow and cord blood. Although the relative distribution of CD34 percentages in each preparation of cells varied widely, on average the percentage of CD34 cells in these different preparations was 0.15%, 0.6%, 2%, 0.45%, 1.68%, and 0.83% respectively. CD34 subset analysis was performed on these cell preparations using multicolor flow cytometry and antibodies to CD33, CD13, CD45RA, CD19, CD71, and CD38. The major differences observed were that bone marrow CD34 cells contain high percentages of CD19+ cells not found in significant quantity in the other cell preparations and cord blood CD34 cells contained a higher percentage of CD38-cells than the other cell preparations. A magnetic bead system was used with anti-CD34 antibody to purify CD34 cells from mobilized peripheral blood apheresis products, cord blood, and bone marrow. Efficient selection with high purities of CD34 cells was achieved with each of the cell preparations. Comparison of colony-forming activity of each of the cell preparations showed cord blood and mobilized peripheral blood to have slightly higher cloning efficiencies than bone marrow with higher numbers of erythroid blast-forming units (BFU-E) also observed in cord blood CD34 cells. Culture of isolated CD34 cells in liquid culture with interleukin-3, stem cell factor, G-CSF, and granulocyte-macrophage GM-CSF showed over a 100-fold expansion in cell numbers after 25 days, with the peak expansion of colony-forming cells occurring between days 11 and 16. Analysis of day-10 cells from these cultures showed them to be predominantly promyelocytes, myelocytes, and metamyelocytes, with cord blood CD34 cultures showing more promyelocytes than peripheral blood or bone marrow and bone marrow showing more metamyelocytes. Comparison of the proliferation of CD34 cells from these different cell preparations showed that cord blood CD34 cells cultured for 10 days averaged an 85-fold increase in cell numbers followed by mobilized peripheral blood CD34 cells, with an average 56-fold increase, and bone marrow CD34 cells, with an average 49-fold increase.

Adult↗
PubMed1995

Semiautomated quantitative detection of loss of heterozygosity in the tumor suppressor gene p53.

The most frequently altered gene in diverse tumor types is the tumor suppressor gene p53. Typically, normal function is inactivated by point mutation of one allele and deletion of the other. Therefore, loss of heterozygosity (LOH) of intragenic polymorphic markers is a strong indication for p53 involvement in a cancerous lesion. This study shows that a highly polymorphic short tandem repeat (STR) within intron 1 of p53 is an excellent marker for quantitative evaluation of LOH in tumor samples, whose multicolor, fluorescently tagged PCR products are analyzed and quantitated on an automated DNA sequencer. The range of error was analyzed in detail. Discrete allelic profiles were obtained following amplification of DNA from microdissected cell samples of patients with urogenital tumors. By calculating qLOH, the relative allele ratio of a tumor compared with healthy tissue, a quantitative expression for the LOH is obtained. PCR-based tumor DNA typing using fluorescent STR primers and automated analysis provides an enhanced level of accuracy and sensitivity required for routine analysis in clinical practice, where large numbers of tumor samples have to be processed.

Alleles↗
PubMed1995

Fluorescence in situ hybridization: powerful molecular tool for cancer prognosis.

We review several aspects of fluorescence in situ hybridization (FISH) technology that demonstrate its breadth and power in detecting and monitoring genetic abnormalities associated with cancers. The clinical utility of FISH in disease management is demonstrated in several examples, including trisomy 8 detection with high specificity and sensitivity in patients with myeloid leukemias; trisomy 12 detection with higher efficiency than conventional cytogenetics in patients with chronic lymphocytic leukemia; assessment of engraftment success, chimerism, and relapse in opposite sex bone marrow transplantation; and correlation of trisomy 7 with survival time in patients with prostate tumors. Advances in FISH technology include multicolor analyses, which permit the simultaneous detection of several genetic abnormalities by using cohybridization of probes labeled with several fluorescent labels or label combinations, and comparative genomic hybridization, a relatively new method whereby a single hybridization can reveal aberrations across the entire genome.

Chromosome Aberrations↗
PubMed1995

Recent advances in fluorescence in situ hybridization (FISH) in hematology.

Fluorescence in situ hybridization (FISH) techniques has been considerably developed over the past years. They have brought new important data when applied to hematological malignancies: studies of aneuploidies on interphase nuclei at diagnosis and in the follow-up of the patients, combination with immunophenotype studies allowing to determine the lineage of the cells studied, simultaneous use of several molecular probes by multicolor hybridization, detection of chromosome rearrangements difficult to ascertain with usual techniques, delineation of the breakpoints in chromosome abnormalities. Examples of these applications are presented and discussed.

Chromosome Aberrations↗