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PubMed1989

[Changes in the pH of leukocyte granules during the storage of preserved donor blood].

Data are presented on pH values in lymphocyte and neutrophil granules, as well as parameters of fluctuating pH changes in individual granules at early terms of storage (up to 72 h), evaluated by cytophotometry with the use of the vital stain neutral red in microspectrophotometer (Univar Reichert, Austria). An average value of pH in donor neutrophil granules was 6.57-0.12 (range 6.21-7.02). At day 3 of storage it decreased at average by 0.18 pH units. Rhythmical changes of pH in individual granules with pH 0.2-0.44 were found on all days of storage. The period of the rhythm lengthened from 3 min on day 1 to 4 min on day 3. This shift as well as acidification of granules during storage reflects their functional deterioration. The storage did not affect pH values in lymphocyte lysosomes which at average were 6.4, 6.5 and 6.35 on days 1, 2 and 3, respectively.

Adult↗
PubMed1994

Scanning microphotometry image analysis of Ha-ras-transformed human breast epithelial cells.

Nuclei of the human breast epithelial cells, MCF-10A, transfected with the c-Ha-ras oncogene, the ras proto-oncogene and the plasmid Homer 6 only, were studied by image analysis after Feulgen staining. This material had been previously used to demonstrate that the experimental insertion of the activated c-Ha-ras oncogene into the DNA of the MCF-10A cells induces their tumoural properties. The ras-transformed nuclei of the MCF-10A cells exhibited differences in chromatin supraorganization in comparison with the nuclei of human breast carcinoma MCF-7 cells, or c-Ha-ras-transformed NIH/3T3 cells and, to a much lesser extent, with those of other MCF-10A transfectants and the non-transfected MCF-10A cells. All MCF-10A transfectants exhibited unravelling of both condensed and non-condensed chromatin, which, however, was less drastic in the ras-transformed MCF-10A cells. It is hypothesized that simultaneous to a general chromatin loosening as a response to foreign transfected DNA, a reverse mechanism may be elicited by ras transformation in the chromatin of the MCF-10A cells. The result in terms of elicited chromatin condensation was not as strong as that promoted in ras-transformed NIH/3T3 cells. Considering that early steps of tumour progression in vitro have previously been assumed to be involved in the ras-transformed MCF-10A cells, the differences in chromatin supraorganization of the ras-transformed MCF-10A cells as compared with MCF-7 cells are probably due to their different tumoural stages plus the putative effect of the transfected DNA vector on the transfected MCF-10A cells.

Breast Neoplasms↗
PubMed1995

[The development of a tissue calcium-detecting apparatus].

Calcium phosphate is deposited in tissues with progression of aging and arteriosclerosis, causing impairments of organ functions. Localization and quantification of tissue calcium phosphate are important for elucidation of the cause of this calcium phosphate deposition. Tissue calcium phosphate has been detected by Von Kossa-Nishiyama staining using the heavy metal substitution technique. By this method, however, the reaction was weak because of the use of sunbeam as the light source, and consistent detection was difficult. Especially, fine calcium phosphate granules that appear in early stages of the pathological changes could not be detected. Therefore, we evaluated a method for preparation of tissue specimens, light source, filter, and staining conditions, and developed a more reliable method for detection and assay of tissue calcium phosphate. The human aortic tissue was fixed with formalin, embedded with paraffin, and cut to a thickness of 4 microns using a glass knife. Solax-XCA100B, an artificial sunbeam lamp with a spectrum nearly identical with that of the sun, was used as the light source. A B filter, which selectively filters out the wavelengths 290nm or less and 800-1,000nm, was used. The appearance of abnormal granules due to excessive reaction could be prevented by eliminating ultraviolet rays with wavelengths of 290nm or less, and burning of the tissue and the appearance of silver particles could be prevented by excluding infrared rays with wavelengths of 800nm or above. Distilled water was continuously dripped into the reaction solution to prevent an increase in the concentration due to evaporation. The reaction showed appreciable progression until after 40 minutes and stopped completely after 80 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)

Aorta↗
PubMed1994

[Influence of foods on the posterior longitudinal ligament of the cervical spine and serum sex hormones].

To investigate the involvement of sex hormones as a potential cause of ossification in the posterior longitudinal ligament (OPLL) of the cervical spine, we experimentally examined the influence of ingested foods on the serum levels of sex hormones and changes in the spinal ligament. Castrated rabbits were administered a sex hormone and raised with special feed. Assay of the serum sex hormone levels, analysis of the carbon and nitrogen isotope ratios in body hairs, and histological studies were carried out. In the group administered soybeans and highly concentrated saline solution, the serum estrogen level was high, the serum testosterone level was low and the delta 15N value of the body hairs was also low. Histologically, a group of newly formed chondroblasts was seen in the posterior longitudinal ligament in the angular region of a narrowed intervertebral disc, and many fibroblasts were detected by assay using a microspectrophotometer (MSP) in this group. These findings suggested that a diet high in vegetable protein, consisting mainly of beans, and foods high in salt may lead to an unbalanced state for sex hormones and cause histological changes in the spinal ligament. This, as well as other general ossifying factors of ligament, may play a role in the etiology of ossification.

Animals↗
PubMed1994

In situ chemical analyses from frozen tissue sections by Fourier transform infrared microspectroscopy. Examination of white matter exposed to extravasated blood in the rat brain.

Fourier transform infrared (FT-IR) microspectroscopy is a powerful technique for obtaining chemical information from tissue sections. Infrared spectra can be collected from individual spots or from several spots along a grid pattern. In the latter example, absorbing bands are interpolated between spectra to generate maps that represent the relative concentrations of chemical groups. In this study, blood was injected into the subcortical white matter of the rat, and the spatial distribution of chemical changes was identified at 2 and 6 days after injection. The absorbances for CH2, H-C = C, C = O, P = O, and HO-C-H were lower at the lesion site compared with the neighboring tissue, which suggests a mechanism of lipid peroxidation. The absorbances for N-H stretch and amide II were greater at the lesion site compared with the surrounding tissue, and the amide II peak was shifted toward lower wave numbers in the affected area. These changes are probably due to the infiltration of blood products, the aggregation of myelin proteins, and/or gliosis in the lesioned tissue. The rate of absorbance change per distance displayed a trend to be more abrupt at 2 days compared with 6 days after injection. We suggest that FT-IR microspectroscopy can be applied to a variety of disease states.

Animals↗
PubMed1994

Chemical and physiological effects of Candida albicans toxin on tissues.

The prevalence of immunologically suppressed patients, including those infected with the AIDS virus, with cancer, and those having had transplant surgery to name a few, has provided an avenue for the rapid proliferation among these patients of the virulent yeast Candida albicans. Previous studies have determined that a potent toxin is produced by C. albicans which may cause extensive tissue damage. The extent of the tissue damage has never been determined, neither has the mechanism been explained. The present work shows that intraperitoneal inoculation of C. albicans produces numerous tumour-like lesions and abscesses on the major organs of experimental laboratory rats. The results demonstrate that damage is caused by the initial release of lysosomal enzymes by the affected tissues.

Acid Phosphatase↗
PubMed1994

[Heterogeneity in gastric carcinoma with special reference to DNA content and mitotic activity: histopathologic differentiation].

DNA ploidy was microspectrophotometrically investigated in 46 patients with gastric carcinoma. Measurements of DNA content and mitotic index (MI) were examined in the mucosal, submucosal, muscularis propria, and serosal layers of tumors, respectively. The frequency of cells with values exceeding hexaploid chromosome (6c) and mitotic counting analysis revealed a significantly higher value in the serosa than in the mucosa. This tendency was not evident in differentiated type adenocarcinoma but was noted in those with the undifferentiated type. There were 37 tumors (80.0%) with the same DNA distribution pattern in every layer of the stomach (homogeneous DNA ploidy). Heterogeneity of DNA ploidy was observed in nine tumors (20.0%). Carcinoma with heterogeneous DNA ploidy manifested a significantly higher incidence of metastasis to lymph nodes than did those with the homogeneous type. Characteristically, there was venous permeation preponderance in the differentiated type and peritoneal dissemination preponderance in the undifferentiated type. This evidence of DNA heterogeneity in gastric carcinoma tissue suggests a possible correlation with metastatic behavior.

Adenocarcinoma↗
PubMed1993

[Prognostic value of nuclear DNA of 106 colorectal tumors, determined with microspectrocytophotometry].

OBJECTIVE: The aim of this study was to look for correlations between nuclear DNA content of colo-rectal tumors, and such clinical parameters as age, sex, location, CEA, histological grade and Duke's stages. EXPERIMENTAL DESIGN: A prospective study is carried out on surgical patients, subjected to standard criteria of radicality. Nuclear DNA content was quantified in tumoral cells by microcytophotometric techniques. PATIENTS: 106 patients with colo-rectal cancer. Patients with colonic perforation, other concomitant neoplasia, non-curative surgery or receiving adjuvant therapies were excluded from the study. Five patients died during the postoperative period and one was lost. RESULTS: Histological grade: 28% G1, 35% G2 and 37% G3. Dukes': 8% A, 40% B, 32% C and 20% D. DNA quantification has rendered 45% as euploid and 55% as aneuploid. There was no statistical correlation between ploidy and location, age, sex or CEA. However, there is a clear preponderance of euploid tumors in G1 (23 vs. 5), while the aneuploid tumors double the euploid ones (24 and 25 vs. 12 and 12) in G2 and G3. A similar result was found comparing ploidy and Dukes: euploid tumors reach 77% both in stages A and B, while they drop to 24% and 14% in stages C and D. It has also been found that euploid tumors show a longer period of survival free of recurrence. CONCLUSIONS: Evidence has been found supporting a prognostic value for tumoral DNA quantification in colo-rectal cancer. A longer follow-up is required to study absolute survival of the patients.

Adult↗
PubMedDate not supplied

Morphologic diagnosis of Hürthle cell tumors of the thyroid gland.

Despite difficulties in differential diagnosis, aspiration cytology offers unique advantages in the preoperative diagnosis of Hürthle cell (HC) tumors of the thyroid gland over immunocytochemistry and micromorphometry, which have not achieved the anticipated success in the diagnosis of HC tumors. Aspiration cytology is comparable, in terms of speed and accuracy, to microspectrometry of DNA content in HC tumors, a technique that is not convenient for routine use. Detailed morphologic analysis of 19 characteristics of the smears and of the cells revealed five parameters adding to the sensitivity, specificity and reliability of routine diagnosis: HC collection in the form of nests, marked or at least moderate anisocytosis and anisokaryosis, multinucleation and emphasized nucleoli. All five criteria are found in > 70% of HC tumors, and the rate of false positives is < 50%. The simultaneous presence of three or four of these five parameters occurred in 70-90% of HC tumors and would lead to a false-positive diagnosis of HC tumors in only 30% of cases. Routine cytologic diagnosis of HC tumors of the thyroid gland may be improved by adoption of these criteria.

Adenoma↗
PubMed1995

Some reflections on the history, and presence of quantitative pathology.

The quantitative approach has always been present in pathology, but this approach is now practiced more often and in more numerous fields than before. Today pathology is applying DNA cytometry, morphometry, stereology, and quantitative immunohistochemistry, all less useful or useless without the quantitative approach. The present fast development of computerized instrumentation has much helped in the spread and application of quantitative principles.

History, 19th Century↗
PubMed1996

[Evaluation of multidrug resistance phenotype on medullary specimens from patients with acute leukemia by determination of nuclear efflux of tetrahydropyranyl-doxorubicin. Approach by confocal laser microspectrofluorometry].

Confocal microspectrofluorometry allows the analysis of fluorescent molecules such as anthracylines in isolated living cells. An optical microscope fitted with a phase-contrast 100 X water-immersion objective enables simultaneous observation of the sample, focusing of the laser beam on the selected cell fraction (nucleus) and collection of the fluorescence emitted from the sample. The resulting intranuclear spectra are interpreted according to a quantitative model of the fluorescence spectra of both free and DNA-bound anthracycline. The intranuclear drug concentration can thus be determined. This technique has been applied to blast cells collected in patients with acute leukemia. Leukemic cells are aspirated from bone marrow, separated by Ficoll sedimentation and resuspended in RPMI-1640 containing 10% fetal calf serum and 200 nM tetrahydropyranyl-doxorubicin (THP-DOX). After one hour, 20 cells are analyzed and the mean nuclear drug content is determined (C1). Cells are then resuspended in the same medium but without anthracycline for 3 hours and the mean intranuclear drug concentration is then also determined (C3). From C1 and C3 the retention rate (RR) is calculated. Firstly, the accuracy of the method was checked. In 4 AML patients, two different samples aspirated on the same day were divided into two portions. Thus, two measurements were made on each one (4 values per patient). Coefficients of variation were satisfactory (4, 6, 12, and 12%). Secondly, blast cells collected in patients with AML and ALL at diagnosis or in relapse were studied. P-glycoprotein (P-gp) and CD34 expression was also studied using respectively immunohistochemistry land flow cytometry. Results obtained from the first 21 patients showed that there was no correlation between RR and either P-gp or CD34 expression. This could result from the efflux of THP-DOX by other mechanisms and/or low sensitivity of the staining technique.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMedDate not supplied

[The detection of the Glm(1) antigen by the indirect immunofluorescence reaction in a quantitative modification].

Quantitative modification of indirect immunofluorescence test permits the detection of Glm(1) antigen in spots on material evidence in much lower concentrations than are detectable by the routine method. Use of the Stat software helps objectively assess the test results after mathematical processing thereof and reliably judge about the presence of the Glm(1) antigen in the tested material.

ABO Blood-Group System↗
PubMed1998

[Potential of the image analyzer "videotest" for performing microphotometric study in cytology].

Precision features of image analyser "Videotest" and possibilities of its applications for cytophotometric investigations have been analysed. It has been shown that a photometric feature is linear at least to optical density 1.1. Cytophotometry of the Feulgen stained rat hepatocyte nuclei has been performed. Coefficients of variation of DNA for 2c, 4c and 8c nuclei are 7.72, 7.65 and 6.50%, respectively, and ratios between average values of these nuclei are 1.00:2.06:4.12. Possible causes of errors of digital television equipment for cytophotometry are discussed.

Animals↗