Cytochemical Changes Research Articles

Histochemical study of colonic cancer in experimental colitis of rats.

A reliable test for premalignant lesions in the development of colonic cancer in chronic ulcerative colitis has been needed. Thus, we designed this cytochemical study, using a model of experimental colitis and colonic tumors induced in Wistar male rats by the feeding of dextran sulfate sodium. The colitis had histologic similarities to ulcerative colitis in man. The percent frequency of polypoid lesions (dysplasia or dysplasia with carcinoma in situ) in the cecum and ascending colon was about 25% at three months and 90% at six months of dextran sulfate feeding. The cytochemical findings by high-iron diamine-Alcian blue staining and Ulex europeus agglutinin binding were chronologically paralleled by histological changes in the colonic mucosa, and the binding pattern of peanut agglutinin was not different between normal and dextran-treated animals. Moreover, some cytochemical changes that occurred during the inflammatory responses were not present in dysplastic or malignant lesions. Thus, the histochemical tests were not useful for detecting of premalignant lesions earlier than by conventional histology. Nevertheless, the dextran sulfate model of colitis in the rat appears suitable for study of cancer development in ulcerative colitis.

Progression of hearing loss in experimental pneumococcal meningitis: correlation with cerebrospinal fluid cytochemistry.

The development of hearing loss and concomitant cerebrospinal fluid (CSF) cytochemical changes in a model of pneumococcal meningitis were examined. Rabbits were injected intracisternally with 10(5) pneumococci. Auditory evoked potentials to clicks and to 10- and 1-kHz tone bursts were recorded hourly; CSF was analyzed every 4 h. Sensorineural hearing loss developed in all animals beginning 12 h after infection and progressed to severe deafness. The onset of hearing loss was preceded by a CSF leukocytosis of > 2000 cells/microL and elevation of CSF protein and lactate concentrations to > or = 1 mg/mL. Temporal bone histopathology showed pneumococci and leukocytes extending from the CSF to the perilymph via the cochlear aqueduct. Hearing loss can develop early in the course of meningitis and is preceded by the abrupt onset of inflammatory changes in CSF. Progression of hearing loss is rapid and proceeds from cochlear base to apex in parallel with the degree of inflammation.

Physical and cytochemical properties of neutrophils activated in situ in the lung during ZAP infusion in sheep.

Increased retention of activated neutrophils in the lungs contributes to endothelial cell injury. However, characterization of the morphological changes that occur in neutrophils during activation in the pulmonary microcirculation has not been fully determined in vivo. Therefore, the present study was designed to determine structural and cytochemical properties of neutrophils in situ in pulmonary arterioles and alveolar capillaries during the infusion of zymosan-activated plasma (ZAP) or plasma (control) in anesthetized sheep. Quantitative morphological methods showed that ZAP infusion caused significant retention of neutrophils in alveolar capillaries [2.19 +/- 0.40 (SD) x 10(8) neutrophils/ml of capillary blood volume] and pulmonary arterioles (1.02 +/- 0.46 x 10(8) neutrophils/ml of arterial blood volume) compared with plasma infusion (1.03 +/- 0.15 and 0.30 +/- 0.10 x 10(8) neutrophils/ml, respectively; P < 0.05). Harmonic mean diameter of ZAP-activated neutrophils in situ (7.19 +/- 0.44 microns) was significantly greater than the diameter of neutrophils in plasma-treated sheep (6.29 +/- 0.17 microns; P < 0.05). Neutrophil cross-sectional area (54 +/- 3 microns2) and volume (248 +/- 27 microns3) in situ in alveolar capillaries were also significantly greater in ZAP-treated sheep than in control sheep (41 +/- 4 microns2 and 184 +/- 9 microns3, respectively; P < 0.05). Similarly, microvascular neutrophils in ZAP-treated sheep were vacuolated and elongated, filamentous actin was redistributed peripherally, and the cells were degranulated. We conclude that during ZAP infusion, neutrophils become enlarged and degranulated in pulmonary microvessels, especially in alveolar capillaries. The structural and cytochemical changes that occur are consistent with the hypothesis that neutrophil activation is accompanied by alterations in neutrophil physical properties, alterations that may facilitate retention and contribute to endothelial cell injury.

Changes of cytochemical properties in the Golgi apparatus during in vivo differentiation of the ameloblast in developing rat molar tooth germs.

The cytochemical changes of the Golgi stacks occurring concomitantly with cell differentiation were examined in ameloblasts of developing rat molar tooth germs using osmium impregnation and cytochemistry with nicotinamide adenine dinucleotide phosphatase (NADPase), thiamine pyrophosphatase (TPPase), and acid phosphatase (Acpase). NADPase, TPPase, and Acpase activities were already present in the Golgi stacks of the inner enamel epithelial cells, the undifferentiated form of the ameloblast: NADPase activity existed in the medial Golgi cisternae, TPPase activity in the trans Golgi cisternae, and Acpase activity in almost all cisternae and strongly in the trans-most cisterna of the Golgi stack. At this stage, however, osmium deposits after impregnation were not observed in the cisterna of Golgi stacks but were present in some small vesicles. These vesicles were located throughout the cytoplasm. Osmiophilic cisternae in the Golgi stacks were apparent for the first time at the stage when the Golgi apparatus developed and migrated to the region distal to the nucleus with the progression of cell differentiation. These findings indicate that the cis subcompartment of the Golgi apparatus was incomplete in the inner enamel epithelial cells with regard to appearance of its cytochemical property, as compared with the medial and trans subcompartments. It is suggested that the cis compartment of the Golgi stack may be completed only in the last stage of the compartmentalized Golgi organization during differentiation of the ameloblast.

Histomorphological and cytochemical changes induced in the liver of goldfish Carassius carassius var. auratus by short-term exposure to lead.

The cellular and molecular effects of experimental lead intoxication (5 mg l-1/24 h and 48 h) were studied in hepatocytes of Carassius carassius var. auratus. Significant lead storage was not detected by atomic absorbance spectroscopy after treatment. Cell morphology appeared only poorly impaired. A decrease of glycogen content was observed. Glycolysis, Krebs cycle and pentose shunt were altered in their enzymatic activities. Lysosomal activities were strongly increased. All enzymatic activities tested were altered 24 h after lead treatment.

Ultrastructural and cytochemical changes in the rat liver during adapation to hypoxia

Ultrastructural and cytochemical changes in the rat liver during adapation to hypoxia

Protection of human, rat, and guinea-pig atrial muscle by mioflazine, lidoflazine, and verapamil against the destructive effects of high concentrations of Ca2+.

In right atrial trabeculae from humans and in left atria from rat and guinea-pig hearts, the protective effects of mioflazine, lidoflazine, and verapamil against the accumulation of cellular calcium were investigated. Two consecutive, cumulative increases in the extracellular calcium concentration, [Ca2+]o (1-25 mmol/l), were induced, in between which the muscles were exposed for at least 30 minutes to solvent or drug. When using solvent in the 30-minute interval, the force of contraction was much lower during the second Ca2+ challenge, while the aftercontractions and the increase in passive tension at high [Ca2+]o tended to be larger. These signs of functional impairment were prevented by exposure to mioflazine or lidoflazine (3 mumol/l each) but not to verapamil (3 mumol/l). Muscles were fixed with glutaraldehyde at the end of the second Ca2+ challenge for morphological and cytochemical examination. After solvent treatment, more than half of the cells were severely damaged, showing cellular edema, contraction-band necrosis, mitochondrial swelling, and nuclear pyknosis; the sarcolemma was devoid of calcium deposits, damaged mitochondria contained either large deposits of calcium or flocculent densities, and in some cells, the cytoplasm was filled with calcium deposits. Following exposure to mioflazine and lidoflazine, but not to verapamil, the number of intact cells after the second Ca2+ challenge was not different from time-matched controls (80-90%). Furthermore, the shifts in cellular calcium distribution were prevented with mioflazine and lidoflazine, whereas verapamil was less effective. There were no species differences with respect to either morphological or contractile changes. In conclusion, exposure of atria to high [Ca2+]o induced similar ultrastructural and cytochemical changes as seen after ischemia-reperfusion induced damage. Indeed, under the mentioned conditions the sarcolemma lost its capacity to exclude Ca2+ after a challenge with high [Ca2+]o and allowed excessive Ca2+ entry. The pathway for this extra Ca2+ remains to be elucidated. L-type calcium channels are probably not involved, since verapamil cannot prevent the Ca2+ overload.

Cytometric and cytochemical study of peritoneal and alveolar macrophages from Yersinia pseudotuberculosis infected ground squirrels (Citellus citellus).

The enzymic activity (succinate dehydrogenase, acid and alkaline phosphatase) of alveolar and peritoneal macrophages as well as leucocytic reaction of ground squirrels infected with Yersinia pseudotuberculosis (I and III serovar) have been investigated in dynamics from the 1st up to the 30th day. The animals infected with III serovar survived only to the 7th day, while those infected with I serovar survived up to the 30th day after inoculation. A massive influx of leucocytes having peak values (100-fold increase) on the 3rd day after infection has been found in the peritoneal cavity of the animals infected with I serovar. Moderate leucocytosis in the blood, and insignificant fluctuations in alveolar macrophage number have been established too. An earlier and higher activation of succinate dehydrogenase in alveolar and peritoneal macrophages from animals infected with III serovar in comparison with those infected with I serovar was observed. No differences in alkaline phosphatase activity of alveolar and peritoneal macrophages have been found between the animals infected with I and III serovar. A correlation has been found between the number of leucocytes and changes in the enzymatic activity of the macrophages. A metabolic transformation was demonstrated typical for different macrophages (peritoneal and alveolar), in the course of this experimental intraperitoneal infection. Obviously, more virulent serovar III of Y. pseudotuberculosis fails to attract leucocytes to the peritoneal cavity sufficiently quickly, so it overcome the local protective mechanisms with consequent systemic cytochemical changes. On the other hand the virulent serovar I attracts leucocytes to the peritoneum and is presumably destroyed by them.

Leaf Abscission in Soybean: Cytochemical and Ultrastructural Changes following Benzylaminopurine Treatment

6-benzylaminopurine (BAP) delays leaf abscission of soybean Glycine max (L.) Merr. Abscission of the distal pulvinus of primary leaves was induced in 12-d-old seedlings or explants by removal of the leaf blade. BAP applied to the cut end of the pulvinus following leaf blade removal delayed abscission. Discoloration of the pulvinus occurred before abscission commenced and the number of grana in chloroplasts within cortical parenchyma cells of the pulvinus decreased over time following leaf blade removal

A cytochemical study of cerebrovascular lesions in mice infected with Plasmodium berghei

Mice with a Plasmodium berghei infection exhibit morphological and cytochemical changes in the blood-brain barrier. Changes in activity and localization of alkaline phosphatase and adenosine triphosphatase, enzymes with important functions in the maintenance of the blood-brain barrier, were observed. Changes in activity and localization of those enzymes in and near the endothelial cells of the microvasculature, concomitant with an increase in pinocytotic activity, and formation of irregular cytoplasmic extensions in these cells, as well as loosening of the basal lamina are indicative of a functional deterioration of the blood-brain barrier in the course of infection.

Partial Cell Wall Lysis and the Resumption of Meristematic Activity in Fraxinus Excelsior Cambium

Cytochemical changes in cambia! cell walls were studied during the transition from rest to mitotic activity in spring. A partial autolysis occurred in the radial walls especially at cell junctions. The lysis was closely associated with a localised decrease in the level of calcium ions bound to the cell walls.

The cytochemical changes and the adherence of neutrophil during cross-linked agarose encapsulated activated charcoal hemoperfusion.

After one hour cross-linked agarose encapsulated activated charcoal hemoperfusion, there was slight increase of total white cells and neutrophil count, but the neutrophil adherence was 16.87 +/- 5.26% and marked lower than that before hemoperfusion (59.91 +/- 5.26%, p less than 0.05). Using glassball hemoperfusion as control, the neutrophil adherence also decreased but not so obvious as the former. Adherence is a cell property that may affect granulocyte margination, diapedesis, chemotaxis and phagocytic function, so the above investigations showed that both the hemoperfusion process and the charcoal itself may reduce the host defence mechanism. There were no change of the peroxidase and alkaline phosphatase activities of neutrophils during one hour cross-linked agarose encapsulated activated charcoal hemoperfusion.

Structure and Ultrastructure of Stomata

Summary Stomata appeared with the first vascular plants in the lower Devonian. The properties of stomatal cell walls play a role in the mechanics and the physiology of stomatal movements. During opening movements, the plasmalemma is convoluted and probably related to ion influx. Microtubules have been shown to be involved in wall building and probably in stomatal functioning. Guard cell mitochondrial membranes undergo configurational changes during stomatal movements and those of open stomata have a larger vacuolar volume with one or few large vacuoles, whereas in closed stomata, there are numerous small vacuoles. In some species, ribosomes are free in the cytoplasm of guard cells of open stomata, whereas in those of closed stomata, they are associated in polysomes. Also, in guard cells of open stomata, nuclei can possess a reticular condensed chromatin and in those of closed stomata, a homogeneous chromatin. These ultrastructural features are associated with the higher synthetic protein and RNA activities of the guard cells when stomata are closed. Ultrastructural and cytochemical changes associated with stomatal movements should be researched in well controlled environmental conditions.

Effects of Secretory Stimulation on the Golgi Apparatus and Gerl of Rat Tracheal Gland Cells

The morphological and cytochemical changes of the Golgi apparatus and GERL of rat tracheal serous cells were studied after in vivo secretory stimulation with pilocarpine. Discharge of PAS-positive secretory granules had started within 30 min and complete discharge was finished within 2 h after stimulation. Reaccumulation of secretory granules became evident at 6 h and the volume of granules was similar to that of unstimulated control cells at 8 h. At 6 h after pilocarpine administration, acid phosphatase (AcPase) activity was markedly increased in GERL and immature secretory granules as compared to uninjected controls and this phenomenon continued 12 h after stimulation. GERL appeared to increase in extent and numerous continuities with immature granules were observed. Thiamine pyrophosphatase (TPPase) activity, normally present in trans Golgi saccules was not altered during the secretory cycle. At 2 h after stimulation, tubular structures of GERL, which also react to AcPase, were seldom observed. At 6 h, these structures were more numerous than those of control cells. These results demonstrate that, in rat tracheal serous cells, pilocarpine induces alteration of only GERL, and reconstitution followed by resynthesis of secretory granules is performed without participation of the Golgi stack.

Multiparametric Evaluation of Retinoic Acid-Induced Terminal Differentiation of Blastoid Cells from Acute Non-Lymphocytic Leukemia Patients in Vitro

Recent findings that retinoic acid (RA) induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line HL-60 in vitro and blast cell maturation in patients suffering from acute non-lymphocytic leukemia (ANLL) prompted an investigation on the ability of this agent to induce terminal maturation in blast cells from ANLL patients in vitro. We tested the ability of RA at 3 x 10(-6) M, 3 x 10(-7) M and 3 x 10(8-) M concentrations to induce differentiation in blastoid cells from 16 patients with ANLL using cytochemical and cytologic parameters, in addition to cytofluorometric methods. Leukemic cells in primary culture from all the patients underwent cytochemical and biochemical changes after treatment with RA. However, the extent of differentiation-positive cell clones (D+ clones) varied from patient to patient. Morphologic maturation was observed in a significant number of bone marrow samples. Leukocyte alkaline phosphatase and NBT reduction ability of cells, which are biochemical markers of granulocytic differentiation, were also significantly increased with a simultaneous decrease in DNA and RNA synthesis (which was estimated using a Phywe ICP-11 impulse flow cytometer).

Cytochemical changes in a human arterial proteoglycan related to atherosclerosis

The cuprolinic blue (CB) staining method has been used to visualize and characterize proteoglycans (PG) in the extracellular matrix (ECM) of normal and atherosclerotic human arteries. Arterial tissues of 13 individuals (1–83 years of age) were obtained by autopsy. For electron microscopic visualization of PGs staining with CB was performed in the presence of a critical electrolyte concentration of 0.3 M MgC1 2. Under these conditions CB selectively interacts with the polysulfated glycosaminoglycan (GAG) side chains of the molecules. Removal of PG side chains by GAG-degrading enzymes prior to CB staining selectively prevented the formation of chondroitin sulfate (CS)-rich and dermatan sulfate (DS)-rich PG-CB precipitates. The DS-rich type of PG is mainly associated with collagen fibrils, the CS-rich type of PG is preferentially localized in nonfibrous areas of the ECM (soluble matrix). When normal arterial tissues are compared with those affected by atherosclerosis quantitative and qualitative changes of PG-CB precipitates are detected. In fibrous plaques a strong accumulation of a large CS-rich type of precipitate close by smooth muscle cells (SMC) and foam cells is observed. In addition, these precipitates are significantly longer in fibrous plaques than in adjacent normal media (116 nm vs. 100 nm; P < 0.001). This alteration is independent of the age of the donor. Small DS-rich PG-CB precipitates associated with collagen fibrils show strong variations in their length, but not a significant tendency towards elongated precipitates in atherosclerosis. The present results demonstrate that ultracytochemical and morphometric analysis are useful in providing information on the diverse types, locations, interactions, and possibly of molecular changes of PGs in normal and atherosclerotic human arteries.

Chronic progressive external ophthalmoplegia. I. A quantitative histochemical study of skeletal muscles.

This study quantitates the major morphological and cytochemical changes in limb muscle biopsies from 37 patients with the syndrome of chronic progressive external ophthalmoplegia (CPEO). The aim was to assess the value of limb muscle biopsy in the diagnosis of this syndrome; to define the myopathological changes and to determine whether there were any specific clinico-pathological correlations. Patients were divided into three clinical groups--11 patients with CPEO with facial and/or limb muscle weakness; 10 with CPEO with facial and/or limb muscle weakness and a positive family history; 16 with CPEO with one or more of the following: pigmentary retinopathy, cerebellar ataxia, pyramidal signs and peripheral neuropathy. The following parameters were measured: the proportions of histochemical fibre types, the muscle fibre areas and the percentage of muscle fibres showing increased oxidative enzyme activity. Pooled results for each of the clinical categories were compared. Statistical analysis of fibre areas and the percentage of fibres with increased oxidative enzyme activity, showed that group 2 differed from the others (p less than 0.05). Patients in group 2 showed the highest incidence of type 1 fibre hypertrophy, type 2A atrophy and the lowest incidence of fibres with increased oxidative activity. Fibre type disproportions occurred in all three groups but the differences were not significant.

Cytochemical changes in hepatocytes of rats with endotoxemia or sepsis: localization of fibronectin, calcium, and enzymes.

Bacterial lipopolysaccharide (LPS) is known to be implicated in the pathogenesis of endotoxemia and septic shock. The liver is the first vital organ to exhibit pathological alterations in shock. The present studies include immunoelectron microscopic localization of tissue fibronectin and cytochemical localization of calcium and enzymes in hepatocytes of animals with LPS-induced endotoxemia or cecal ligation-induced septic shock. The results showed increased staining of fibronectin in the basal (perisinusoidal) surfaces and in the cisternae of rough endoplasmic reticulum and the Golgi complex of hepatocytes in rats with endotoxemia or septic shock. Intracellular calcium content was significantly increased in the LPS-treated or septic rats. Calcium pyroantimonate precipitate was deposited predominantly on the outer surfaces of the RER of hepatocytes. In addition, diminution or depletion of glycogen, reduction of catalase-containing peroxisomes, increase of G-6-Pase activity, and depletion of cytochrome c oxidase in many mitochondria were also observed in hepatocytes of experimental animals. The overall results suggest that LPS stimulates: (a) hepatic synthesis and secretion of fibronectin; (b) uptake of calcium by hepatocytes; and (c) G-6-Pase activity. LPS treatment also leads to reduced numbers of peroxisomes and depletion of cytochrome c oxidase.

Cytochemical studies on the effect of intraperitoneal and oral administration of a traditional chinese medicine(sho-saiko-to) on the D-galactosamine-induced hepatic injury of rats.

The effect of “Sho-saiko-to” (TJ-9), a traditional Chinese medicine, on the D-galactosamine (D-Gal)-induced hepatic injury of rats was estimated from morphological and cytochemical aspects, comparing the methods of administration of TJ-9.A single administration of D-Gal to rats resulted in a hepatic injury with some morphological and histo-cytochemical changes. Both light and electron microscopy showed a disorder in the hepatic lobular structure and configuration of hepatic cords, retention of lipid droplets in hepatocytes, infiltration of inflammatory cells, a decrease in the number of sinusoidal microvilli, dilatation of bile canaliculi, etc. Cytochemical observation showed a remarkable decline of activities of 5′-nucleotidase (5′Nase), a plasma membranous enzyme, and glucose-6-phosphatase (G6Pase), an endoplasmic membranous enzyme, in the liver cells injured by D-Gal. However, in the rats receiving prophylaxis or therapeutic treatment with TJ-9 by intraperitoneal or oral administration, neither severe morphological changes nor a remarkable decline of the enzyme activities was observed. These results suggest that the prophylaxis with TJ-9 prevents the occurrence of hepatic injury, and therapeutic treatment improves the pathological changes induced by D-Gal. Comparing the methods of TJ-9 administration to the rats, intraperitoneal administration was more effective than oral on the hepatic injury.Enzyme cytochemical changes in the liver could be regarded as a useful hallmark for estimating the effect of a drug and the degree of hepatic dysfunction, as well as structural changes in the liver tissue. The present study provided morphological and cytochemical evidence supporting the fact that TJ-9 is efficacious in the treatment of human hepatitis.

Cytochemical study of the Golgi apparatus and related organelles of the secretory ameloblasts of rat molar tooth germs cultured with and without colchicine

The Golgi apparatus and Golgi-associated endoplasmic reticulum lysosome (GERL) were examined in the ameloblasts with a cytochemical marker, osmium impregnation, and two enzyme markers, thiamine pyrophosphatase (TPPase) and acid phosphatase (ACPase). In control cultured germs, osmium deposit appeared in one to two immature side cisternae of Golgi stacks; TPPase activity was restricted in a few mature side cisternae and condensing vacuoles. ACPase activity existed in the GERL and, sometimes, in the mature side-cisternae and condensing vacuoles. These findings show that Golgi stacks of ameloblasts consist of several distinct compartments. In colchicine-treated tooth germs, there were morphological and cytochemical changes in both Golgi stacks and GERL. The Golgi apparatus was fragmented and its stacks were scattered throughout the supranuclear region. In some stacks, the number of osmium-positive cisternae was greater than normal; in others they were absent. TPPase and ACPase activity was absent or diminished. These findings suggest the importance of microtubules in the organization of Golgi complex and GERL in the secretory ameloblast.