[Gastroenterology and hematology].
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Diagnosis, Differential↗
Biomedical subjects
S Witte
Publications and source records attributed to S Witte.
The radial protein concentration profile in the interstitial space of the rat ileal mesentery.
The radial distribution of the protein concentration in the interstitium between arteriolar and venular vessels of the ileal mesentery of the rat was examined. Protein mass was determined by means of uv ultramicrospectrophotometry (UMS) and the relative volume distribution by means of fluorescence microscopy (FM) using the Na fluorescein and FITC-dextran (10,000 mol wt). UMS revealed gradients for protein mass from the vessels out into the interstitial space. FM showed a uniform distribution of fluorescence in the interstitium between the vessels. A gradient for protein mass without a gradient for volume distribution signifies the presence of a concentration gradient for protein in the interstitial space. The protein concentration across the arteriolar wall drops from 5.4 +/- 0.24 (SD) to 2.6 +/- 0.65% and across the venular wall from 5.4 +/- 0.24 to 3.3 +/- 0.43%. From the perivascular site the protein concentration declines exponentially reaching a minimum average interstitial concentration of 1.6 +/- 0.56%. Minimal protein concentration occurred at a point 37 +/- 6.4% of the 295 +/- 37 micron distance from the arteriolar to the venular vessels. In view of this distribution, it is unlikely that lymph or direct samples of interstitial fluid are representative of the perivascular protein concentration.
Animals↗
Measurement of the optical thickness of transparent tissue layers.
The method of Duc de Chaulnes was employed to determine the mechanical and optical thickness, as well as the refractive index, of transparent tissue layers in living specimens. To this end the reproducible accuracy of the method and its dependence on the adjustment in focusing and on the numerical aperture of the objective was evaluated on test specimens using the procedures of transmitted light, phase-contrast (PC), and differential interference contrast (DIC) microscopy. The best working conditions were then applied to the actual measurements.
Animals↗
[Incidence and significance of the gastric mucosal constellation in pernicious anemia].
1977 cases with gastric complains but without localized findings like ulcer or cancer were investigated by gastroscopy with biopsies of corpus and antrum ventriculi. We found in 5.5% an isolated mucosal atrophy of the gastric corpus (mean age 67.2 years compared to 59.4 years in cases with normal mucosa). 0.7% showed an atrophic gastritis of the antral mucosa, 0.9% an atrophic gastritis of the body and the antrum. Most of the cases with an isolated atrophy of the gastric body gave findings of hematologic disturbances. In all cases with isolated atrophy of the gastric body, including localized findings in the stomach, we found in 10.8% neoplastic processes, in another 3.9% hyperplasiogenic polyps. It seems to us of clinical importance to evaluate the gastric histology of the antral and body mucosa routinely because of the high incidence of pernicious anemia-like gastric atrophy and its relation to gastric neoplasias.
Adult↗
Analysis of the wall permeability of blood vessels in the rat mesentery.
The transport of fluorescent tracer molecules of various molecular weights (MW 340-300,000) within the blood vessels and through the vessel walls into the perivascular tissue of the rat mesentery was studied microscopically. Using a highly sensitive TV-tube the fluorescence intensities were transformed into video signals and recorded for subsequent analysis. The results show that small molecules, such as the water-soluble dye fluorescein-sodium ( F1Na ) (MW 340) can pass through the wall of all blood vessels with minimal delay, whereas the passage of large molecules, such as the serum proteins (MW greater than 60,000), depends very strongly on the diameter of the vessels and on the medium in which the mesentery is embedded during the observation. In this respect, no difference was registered between vessels of the arterial and of the venous part of the microcirculation. The large serum protein molecules moved through the blood vessel walls at specific leakage points. The small dye molecules, however, traversed the wall uniformly along the entire length of the blood vessel. The small-molecule behavior could be described by a passive diffusion model with a cylindrical diffusion source.
Animals↗
The role of blood coagulation in capillary permeability Vitalmicroscopic contributions.
It is our concept that the blood coagulation and fibrinolytic systems contribute to capillary permeability. By means of fluorescent intravital microscopy we measured in situ the permeability of proteins and fluorescent tracers through the walls of various kinds of microvessels (exposed rat mesentery). Fluorescent dyes applied intravenously pass across the walls of all kinds of microcirculatory vessels into the perivascular tissue in animals having a state of hypocoagulemia in a shorter time than in normals. Fluorescent-tagged fibrinogen accumulates at the vascular wall, especially at the inner lining of venules, thus marking free receptor sites for fibrinogen at these places. Blocking the fibrinolytic activity of the blood augments the fibrinogen deposition. Pretreatment by heparin does not prevent it, which speaks against the fibrinogen-fibrin coagulation during the accumulation phenomenon. In contrast to this behavior of fibrinogen, albumin, gammaglobulin, antithrombin III, plasminogen never accumulate at the vessel wall but pass through the vascular wall, mostly at venules. The speed and amount of the transvascular passage of these proteins depends on their molecular weight. Fibronectin shares the same receptor sites at the inner lining of microcirculatory vessels as fibrinogen. It seems possible, therefore, that both proteins, fibrinogen and fibronectin, interact with each other at the endo-endothelial cell border. A low molecular weight polypeptide fraction of factor VIII accumulates at the vascular wall similar to fibrinogen. At the same time it decreases the capillary permeability for serum proteins.
Animals↗
[Cytologic diagnosis of the upper gastrointestinal tract].
Three examples are given of new developments in applying cytological procedures in the diagnosis of gastro-intestinal tract disease. At first chinese results in early diagnosis of esophageal carcinoma are described. Then cytological diagnosis of pathological changes in the region of the Vater's ampulla are discussed. Finally trends of quantitative cytology are elaborated upon. We had the opportunity to examine cytological preparations from China, which had been collected in different regions with particularly high or low incidence of esophageal carcinoma. Preparations from regions with high incidence of carcinoma showed a sequence of lesions of esophageal cells starting from chronic esophagitis going over squamous cell hyperplasia and dysplasia of moderate and severe degree leading up to typical cancer cells. Cytological examination of cells collected in the region of Vater's ampulla by brushing technique may yield results demonstrating the presence of benign or malignant tumors. Cytological examinations of pancreatic juice collected after secretin stimulation may give hints in regard to the presence of pancreatitis or pancreatic carcinoma, and examination of bile collected endoscopically from the choledochus may allow diagnosis of cholangitis or primary bile duct tumors. Our own experiments in using quantitative gastroenterological cytodiagnostic procedures are described. They are based on single cell and continuous flow cytofluorometry of DNA in material collected during endoscopy by brushing technics from the stomach. The same material was examined with monochromatic UV microscopy, which allows electronic analysis with high resolving in power of absorption patterns of undyed cell nuclei.
Ampulla of Vater↗
Surface phenomena of coagulation factors in vivo, observed by fluorescence intravital microscopy.
The microcirculatory blood vessels of the exposed rat mesentery have been observed by incident fluorescence microscopy using various optical systems as well as image-intensifier devices and fast-scanning fluorometry techniques. As fluorescence-labeled proteins, albumin, gamma-globulin, fibrinogen, fibronectin, factor VIII-derived polypeptides, and antithrombin III have been injected intravenously. In some experiments two different-labeled proteins were administered consecutively. Whereas albumin and gamma-globulin did not accumulate at the inner surface of the microcirculatory blood vessels , fibrinogen and fibronectin showed a significant affinity to the vessels inner lining, mostly in venular vessels. Both substances accumulated at the same places of the microcirculatory endothelium. Blocking the fibrinolytic system of the animal increased the deposition of both substances. Factor VIII-derived polypeptides accumulated at the vessel inner surface in a different way. Antithrombin III did not show any affinities to the vascular wall. A receptor at the endothelial cell surface for both fibrinogen and fibronectin is postulated.
Animals↗
The measurement of density and its significance in blood rheology.
When viscoelasticity is measured with an oscillating capillary rheometer, the density of the fluid under study is an influencing factor (inertia). In the particular case of blood rheology hematocrit is also of interest. It can be measured more exactly on the basis of the density of blood and blood plasma than by conventional methods using a hematocrit centrifuge or Coulter counter, as these give too high readings when there is an increased tendency towards erythrocyte aggregation and reduced erythrocyte flexibility. Due to the double significance of blood density for blood viscoelasticity, an oscillating capillary rheometer and a new type of density-measuring device were attached to a computer in such a way that simultaneous evaluation is possible. The density-measuring device is a U-tube that is set into oscillation electronically. The duration of the oscillation is an indication of the density of the fluid in the U-tube.
Blood Physiological Phenomena↗
The endoendothelial lining as studied by a fluorescent labeling technique in situ.
The microcirculatory blood vessels of the exposed rat mesentery were observed by intravital microscopy using incident fluorescence excitation and transmitting bright field illumination. Fibrinogen conjugated with the fluorescent dye Fluorescein-Isothiocyanate and injected intravenously was found to accumulate at the vessel wall mostly at the inner lining of venules. The accumulation did not occur in an uniform manner, but appeared as circumscribed streaks, nets and bands in some areas whereas other parts of the microcirculation remained free of any fibrinogen wall adherence. Heparin pretreatment did not influence the phenomena. Blocking the fibrinolytic activity of the blood increased the fibrinogen deposition. Fibronectin labeled with a second fluorescent dye, Rhodamin B-Isothiocyanate, could be differentiated from fibrinogen in the same animal. Both proteins accumulated at the same places of the vessels inner lining. Since albumin and gamma globulin did not show any affinities to the vessel wall, we postulate receptors for fibrinogen and fibronectin at the endoendothelial surface of the microcirculatory endothelial cells.
Animals↗
The endoendothelial lining as studied by a fluorescent labeling technique in situ.
The microcirculatory blood vessels of the exposed rat mesentery were observed by intravital microscopy using incident fluorescence excitation and transmitting bright field illumination. Fibrinogen conjugated with the fluorescent dye Fluorescein-Isothiocyanate and injected intravenously was found to accumulate at the vessel wall mostly at the inner lining of venules. The accumulation did not occur in an uniform manner, but appeared as circumscribed streaks, nets and bands in some areas whereas other parts of the microcirculation remained free of any fibrinogen wall adherence. Heparin pretreatment did not influence the phenomena. Blocking the fibrinolytic activity of the blood increased the fibrinogen deposition. Fibronectin labeled with a second fluorescent dye, Rhodamin B-Isothiocyanate, could be differentiated from fibrinogen in the same animal. Both proteins accumulated at the same places of the vessels inner lining. Since albumin and gamma globulin did not show any affinities to the vessel wall, we postulate receptors for fibrinogen and fibronectin at the endoendothelial surface of the microcirculatory endothelial cells.
Adsorption↗
[Intra- and extravascular distribution of coagulation proteins. Interaction with the vascular wall].
Blood proteins specific for the clotting process show the same distribution behavior into the extra vascular space like other plasma proteins. They participate in the macromolecular transport through the vessel wall into the interstitial space and the lymph fluid. The reported results were obtained by vital microscopic investigations of mesenteric vessels of rats using different quantitative techniques with labeled proteins. Fluorescein labeled fibrinogen, antithrombin III, and plasminogen in this technique behave differently according to their molecular weight differences. In humans the extravascular distribution of clotting factors was studied using the model of cantharides skin blister. Some clotting factors were found to be diminished to a different degree pointing to clotting phenomena during transport. - In contrast to most other plasma proteins fluorescein labeled fibrinogen is found to have an affinity to blood vessels, as can be shown using the vital microscopic technique and rat mesenteric vessels. With this, one can detect areas at the interendothelial cell borders of small veins where the fibrinogen turnover primarily takes place. Similar affinities have been detected with fibronectin and a polypeptide from F VIII digest. The results are interpreted as a morphological hint at the functional connection between blood coagulation and the permeability of vessels for macromolecules.
Animals↗
Comparison of ftorafur with 5-fluorouracil in combination chemotherapy of advanced gastrointestinal carcinoma.
The aim of the study was to compare the combination 5-FU-carmustine with ftorafur-carmustine in the treatment of advanced gastrointestinal cancer. To this end, a prospective, multicenter, randomized trial was initiated. Part I of this trial showed that similar response rates can be obtained with 5-FU-carmustine and ftorafur-carmustine in 109 patients (32.7% versus 26.3%). However, median survival was better in patients treated with 5-FU-carmustine (307 days versus 163 days). Part II of the trial revealed that neither a higher dosage of ftorafur (2 g/m2/day X 5 days) nor the addition of vincristine to both regimens changed the previously obtained results significantly. Again, median survival was found to be better in patients treated with 5-FU combination chemotherapy (304 days versus 144 days). Both the 5-FU and the ftorafur combination were tolerated reasonably well. The results suggest that combination chemotherapy including 5-FU is superior to ftorafur at the applied dosages in terms of survival.
Adult↗
The affinity of fluorescent labeled fibrinogen to the vessel wall as seen by vital microscopy.
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Animals↗
[Transcapillary exchange of micro- and macro-molecules (author's transl)].
The permeability of the terminal vascular bed for micro- and macromolecules influences the hemorheological behaviour of the blood in various ways: 1. Water and water-soluble micromolecules leave the circulating blood at the arteriolar part of the microcirculation, thereby increasing the colloid osmotic pressure as well as the hematocrit downstream towards the capillaris and venules. 2. Water enters the blood from the tissue preferentially at the venules. This process may be disturbed easily by variations of the hydrostatic pressures with serious consequences for the streaming blood and for the tissue. 3. The permeability of macromolecules takes place mostly as the venous part of the capillaries. It depends on the molecular size and will be governed by the vessel wall itself. The coagulation system influences these processes. Together with the reabsorption of water from the tissue the protein movements through the capillary wall stabilize the hemorheological factors within the venules. The venules are the most vulnerable part of the circulation. 4. The movements of plasma proteins within the interstitial tissue are caused by concentration gradients. Finally the lymph collects the extravascular proteins. Within both compartments, th extravascular space and the lymph, various microrheological phenomena are to be expected.
Blood Coagulation↗
The diagnostic significance of flow cytometric nuclear DNA measurement in gastroscopic diagnosis of the stomach.
Cell material obtained from 161 patients by gastroscopic aided brushing was compared cytologically and DNA flow cytometrically and evaluated by mathematical discriminant analysis. Whereas all but one of the cytologically confirmed malignant cases were also positive (malignant) in flow cytometry, the DNA determination failed in 6 of the 19 cytologically suspicious cases, having been classified as negative (benign) by cytometry. Out of all cytologically unequivocal malignant and suspected cases, 18% (7 cases) were false-negative in flow cytometry. The false-positive rate was 39% (47 cases). 5% of the material was classified as inadequate for diagnosis (8 cases) by flow cytometry. The false-negative rate of flow analysis in cytologically suspicious cases is apparently caused by a less pronounced increase of DNA content in the cell nuclei of these cases. Reactive gastric mucosa alterations such a inflammations, erosions or ulcers exhibited increased cell nuclear DNA. According to these results, DNA determination can be used as an automated diagnostic procedure for tumour detection only in combination with other measuring criteria.
Cytological Techniques↗
[Automated blood cell differentiation by the Hematrak].
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Blood Cell Count↗
Microphotometric techniques in intravital microcirculatory studies.
Intravital microscopic techniques were developed in order to measure plasma proteins: (1) circulating in the streaming blood of microcirculatory vessels of the exposed rat mesentery, (2) permeating the vessel wall, and (3) moving through the perivascular interstitial tissue. By means of the ultramicrospectrophotometry, the absorbance of proteins containing aromatic amino acids at 280 nm is measured. Absolute amounts of proteins in the range of picograms are measured in a circular measuring field with a diameter of 17 micrometers. Problems of calibration and of unspecific light scattering are discussed. The scanning microfluorometry makes use of the fluorescent labelling of the plasma proteins. Interstitial tissue with a capillary vessel containing labelled proteins is measured at high speed. A motor-driven object stage synchronized with a measuring system uses scanning lines to traverse the tissue. The data are then processed by a computer. In this way we get continuous quantitative information about the dynamics of protein permeability, starting from time zero. The measuring of the fluorescence intensity and the speed of the fluorometric system are further improved by a computerized 10 kHz scanning device.
Animals↗