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PubMedDate not supplied

Interaction of iron polymers with blood mononuclear cells and its detection with the Prussian Blue reaction.

An inhibitory effect of iron salts on various immune functions in vitro has been reported in several laboratories during the last few years. This study confirms and extends those observations by showing that iron citrate inhibits the mitogen-induced (PHA, Con A and PWM) lymphocyte proliferation. Such inhibition is observed in the presence of ferric citrate with a metal-to-ligand molar ratio of 1:1 but not with ferric citrate with metal-to-ligand molar ratio 1:20 in which the formation of polynuclear iron complexes is prevented. Increasing the concentration of serum in the culture medium diminished the inhibitory effect of 1:1 ferric citrate. Using the Prussian Blue reaction the presence of ferric iron was observed on the cell surface. It is proposed that the deposition of polynuclear iron complexes on the lymphocytes membrane is one of the possible factors determining the iron inhibitory effect.

Cells, Cultured↗
PubMed1988

Stainable bone iron in undecalcified, plastic-embedded sections. Occurrence in man related to the presence of "free" iron?

Iron demonstrable with the Prussian blue reaction at the osteoid/mineralized tissue interphase (osteoid seam) of trabecular bone was observed in only 2.3% of a total of 1536 conventionally fixed and processed, undecalcified, plastic-embedded biopsy specimens taken from the iliac crest of patients for various diagnostic purposes. In marked contrast, clearly stainable bone iron was noticed in all 4 biopsy specimens from the iliac crest and in 11 of 15 vertebral bone fragments obtained at autopsy from individuals with verified primary or secondary hemochromatosis. Findings, including results obtained in vitro, suggest that a positive Prussian blue reaction at the surface of trabecular bone signals the presence of low-molecular-weight ("free") iron, which can bind to the osteoid matrix directly, ie, without the help of osteoblasts. Stainable bone iron may thus be a useful criterion for early detection of hemochromatosis and other types of potentially toxic iron overload.

Adolescent↗
PubMed1988

[The pathological study of enterosiderosis in guinea pigs].

Enterosiderosis in both SPF Hartley guinea pigs and vitamin C-deficient animals of the same strain were studied by light and electron microscopy. Enterosiderosis was detected in all animals in the present study. Macrophages, inclosing yellowish-brown pigments and erythrocytes, appeared in the lamina propria of the intestinal mucosa, mainly in the cecum. These pigments in the macrophages were positive for Prussian blue, PAS and the Nile blue reaction. Residual bodies containing highly electron-dense ferritin-like particles, lipofuscin granules and debris of phagocytized erythrocytes were found by electron microscopy in the macrophages. In vitamin C-deficient guinea pigs, the number of macrophages, including the same above pigments, appeared in the lamina propria of the intestinal mucosa, and there was severe enterosiderosis. In the absorptive cells of the intestinal mucous membrane, granules positive for the Prussian blue reaction appeared only in the duodenum. These findings strongly suggest that the pigments in the macrophages in enterosiderosis of the guinea pigs were mixtures of iron and lipofuscin granules and that the iron is derived from erythrocytes phagocytized by macrophages in the lamina propria, but not from iron absorbed by epithelial cells.

Animals↗
PubMed1978

Ultrastructural localization of nonheme celluar iron with ferrocyanide.

The Prussian blue reaction was evaluated at the ultrastructural level as a cytochemical method to identify ferric and ferrous iron in rat bone marrow and splenic macrophages. Satisfactory tissue preservation and staining were achieved after fixation for 1 hr in 3% glutaraldehyde and exposure for 30 min to Perls's ferrocyanide solution before routine osmication and embedding. The acid ferrocyanide solution formed cuboidal and irregular electron-opaque deposits which localized ferric iron in the macrophage siderosomes and hyaloplasm. When thin sections were directly stained with the acid ferrocyanide, the stain deposits were much less distinct. The size and number of cytes exhibited sparse evenly distributed stain deposits. Several cells displayed abundant precipitates on the inner surface of the plasmalemma. Prussian blue precipitates were occasionally seen in mitochondria and nuclear euchromatin. Although osmium tetroxide post-fixation improved tissue preservation, it did not enhance the density of the ferri-ferrocyanide precipitate. The ferrocyanide solution yielded cuboidal deposits also in clots impregnated with ferritin, and electron diffraction analysis confirmed the symmetrical crystal structure of these stain precipitates. Smaller irregular precipitates were formed in clots impregnated with FeCl3, or Fe2 (SO4)3 solutions, despite the equally interpreted as indicating that the iron hydroxide core or protein structure of ferritin and hemosiderin contributed to the formation of the ultrastructurally evident cuboidal precipitates, but were not necessary for the formation of a colored reaction product. The acid ferrocyanide solution failed to stain clots formed in FeCI2, CuCI2 or CuCI solutions. Staining with a ferricyanide solution identified only sparse foci of ferrous iron in some siderosomes. This study demonstrates that the Prussian blue reaction can be used ultrastructurally to localize iron cations bound to some nonheme iron binding proteins.

Animals↗
PubMed1979

Ferrocyanide staining of transferrin and ferritin-conjugated antibody to transferrin.

To evaluate the ultrastructural distribution of transferrin on the surface of L1210 ascites tumor cells, we used ferrocyanide to stain ferric iron (Prussian blue reaction) in transferrin, as well as in ferritin conjugated to antibody that was immunologically attached to the transferrin. Small deposits averaging 5 nm in diameter identified transferrin iron, whereas large cuboidal deposits averaging 50 nm in diameter stained ferritin conjugated-antibody that was bound to both transferrin and apotransferrin on the cell surface. The ability of transferrin to deliver iron to ascites tumor cells was confirmed by kinetic studies of transferrin labeled with 59Fe and 125I. These preliminary results are consistent with release of transferrin iron at the cell surface and demonstrate additional uses for ferrocyanide in ultrastructural cytochemical techniques.

Animals↗
PubMed1978

Ultrastructural cytochemistry of iron absorption.

Conventional ultrastructural autoradiographic and morphologic studies of the duodenal mucosal cell have generally corroborated physiologic observations of iron absorption, but such methods have limited resolution and fail to distinguish ferric and ferrous iron. This study describes the application of the Prussian blue reaction as an electron microscopic cytochemical stain to the investigation of inorganic iron absorption in iron-deficient, normal, and iron-loaded rats. Ferrous iron is converted to ferric iron at the microvillus membrane. Subsequently intraepithelial ferric iron appears bound to a non-heme acceptor substance in microvilli and later appears as small non-membrane-bound stain deposits which are concentrated in the apical cytoplasm. The appearance of larger stain deposits in the lateral intercellular spaces, in the basal extracellular spaces, and along the intraluminal and extraluminal outer plasmalemma of adjacent endothelial cells of the lamina propria suggests passage of iron from epithelial cells through the lamina propria to blood vessels. The extreme sensitivity of the method compared with simultaneous ultrastructural autoradiographic techniques is demonstrated and suggests usefulness of the method in further studies of iron metabolism.

Animals↗
PubMed1984

[Blood destruction in pulmonary alveoli: signs of vitality and determination of survival time].

Lungs from 26 cases were examined in which blood was present as a result of a gunshot wound, a stab wound, or aspiration. Signs of vitality and of a time-dependent reaction sequence were evaluated to determine survival time. Only those morphologic criteria were considered that could be obtained on paraffin sections. In addition to H & E staining, siderin was identified with the Prussian blue reaction and the activities were determined of tartrate-resistant acid phosphatase, as macrophage marker, and naphthol AS-D chloracetate esterase, as granulocyte marker. The following criteria were evaluated: granulocyte emigration, erythrocyte adherence to the surface of macrophages, macrophage ingestion of erythrocytes, and determination of siderin as indicator of intracellular erythrocyte digestion. Adherence was also observed in those cases that did not survive. The initial sign of vitality was granulocyte emigration, which was observed for the first time after a survival time of 5 min. Erythrophages were found after a survival time of 30 min at least, siderophages after 17 h at the earliest. Literature dealing with vitality and age determination, as well as the pathogenesis of the reaction sequence, is discussed.

Adolescent↗
PubMed1988

The mechanism accounted for the silver staining "fast technique" development and its histochemical meaning: I. The correlation between the silver staining intensity and the tissue bound ferric ion and some other tissue histochemical reactivities.

The mechanism accounted to accomplish the silver staining "fast technique" on tissues sections was studied towards the correlation among histophotometric measures concerning the silver staining intensity and the intensity provided by some histochemical reactions performed on spleen and liver sections from rats and pigs. By treating previously these histological sections with thioglycolate or oxalate solutions in progressive concentrations and afterwards subjecting them to a silver staining "fast technique", it was demonstrated that the silver staining intensity decreases proportionally to the thioglycolate or the oxalate solution concentration. The regression line of the silver staining intensity on the thioglycolate or the oxalate solution concentration was established, as well as its regression coefficient. On the other hand, on histological sections previously subjected to the thioglycolate or the oxalate treatment, some histochemical techniques were performed and the histophotometric measures concerning the intensity of each histochemical technique used were taken and its regression line, as well as its regression coefficient were established. By comparing the silver staining intensity regression line (or its regression coefficient) with the regression lines established for the histochemical techniques used, it was tested the correlation between the silver staining and the reactivity of some reactive groups contained into the tissues. In this manner, the influence of such reactive groups on the silver staining development was tested. The results show that there is no correlation between the silver staining intensity and the reactivity degree of the reducing groups (-SH- and carbonyl group), the 1-2-glycolic group, as well as of some protein reactive groups (phenol, imidazole, carboxylic groups). On the other hand, taking into account the respective regression lines and its regression coefficient, the comparison between the silver staining intensity and the Prussian blue reaction intensity shows a close relationship between them. This finding suggests that for the silver staining "fast technique" the tissue bound ferric ion accomplishes a very important role and can be accounted for its mechanism and its histochemical meaning.

Animals↗
PubMed1982

The structural correlate of saltatory conduction along the Mauthner axon in the tench (Tinca tinca L.): identification of nodal equivalents at the axon collaterals.

The spiny collaterals of the Mauthner axon were reinvestigated in the tench (Tinca tinca L.) with the electron microscope and special staining procedures. These collaterals, as demonstrated by intraaxonal labelling with lucifer yellow, are more or less regularly spaced (100-300 micrometers) and make synaptic contacts with processes of spinal motoneurons and interneurons. The unmyelinated tips of the collaterals are further characterized by the following structural features: (1) an electron-dense undercoating of the axolemma, (2) a positive Prussian blue reaction of the inner surface of the axolemma following ferric ion-ferrocyanide staining (Waxman and Quick, '78a), (3) expanded extracellular spaces which react specifically to inorganic phosphate, metallic ions, and diaminobenzidine. All these properties are known to be shared by the axolemma of central and peripheral nodes of Ranvier. Previous studies from this laboratory have shown that the nerve impulse is propagated along the Mauthner axon in a saltatory mode. Since classical nodal gaps could not be identified within the myelin sheath of this giant fiber, it is concluded on the basis of the present findings that the unmyelinated tips of the spiny collaterals represent nodal equivalents, and thus provide the morphological substrate for the saltatory propagation of the nerve impulse along the Mauthner axon. The typical latency steps, as demonstrated in the latency plot of the longitudinal current signals (Greeff and Yasargil, '80), and the distances between the identified membrane specializations at the axon collaterals are consistent with this conclusion.

Animals↗
PubMed1987

[Hemosiderin findings in the liver, spleen and lung in newborn infants and infants].

The content and distribution of hemosiderin in the liver, spleen and lungs of stillborn children, newborns and infants were determined histologically by Perl's method (Prussian blue reaction). We describe a study of 59 deaths up to the age of 2 years. The hemosiderin content depends on age and decreases with increasing age. No relevant differences were found in cases of sudden infant death syndrome (SIDS). In the liver hemosiderin is localized preponderantly in the periphery of the lobules. The spleen shows a diffuse siderosis of the red pulp. Minimal intrapleural, septal and perivasal depositions of hemosiderin can be found in the lung tissue. The significance of these findings is discussed.

Female↗
PubMed1989

A new method of the immunohistochemical detection of cellular antigens for light and electron microscopy.

A new immunohistochemical method for light and electron microscopy of tissue- and cell-specific antigens by using ferric colloid-labeled antibody is presented. The antibodies labeled with the cationic cacodylate ferric colloid are stable and bind specifically to the target antigens to show clearly the site of antigens in tissue sections and on free cells by Prussian blue reaction for light microscopy and by the specific figure of electron opaque ferric colloid particles for electron microscopy. The staining procedure is very simple and it gives clear picture. So the method will be of beneficial for general laboratory use in immunohistochemical researches.

Animals↗
PubMed1985

Fine-granular cationic iron colloid. Its preparation, physicochemical characteristics and histochemical use for the detection of ionized anionic groups.

In order to obtain distinct and reliable information concerning the localization of ionized anionic groups in tissues, fine-granular cationic ferric hydroxide colloid solution (Fe-Cac-f) was newly devised. This can be obtained by boiling a mixture of ferric chloride and ammonium cacodylate solutions. The colloid particles of Fe-Cac-f are about 1.0 nm in size, i.e., one-fifth of the size of ferric cacodylate colloid (Fe-Cac; Seno et al. 1983a). As with Fe-Cac, Fe-Cac-f particles in the pH range of 1.6-7.6 carry a positive electric charge, but the latter show a better permeation of tissues. Using the Prussian blue reaction, Fe-Cac-f gives a distinct deep-blue color and can be used for the detection of anionic groups of acid mucopolysaccharides and proteins by light microscopy. It is also useful for detecting the exact sites of ionized anionic groups in deep tissue areas using electron microscopy.

Animals↗
PubMed1979

Iron-induced atypical meningioma cells?

Polymorphous tumor cells with bizarre giant nuclei are characteristic of the so-called atypical meningioma. Some of these cells are iron-positive. Meningioma cells apparently are able to store iron excessively. It is discussed whether an extremely high intracellular iron uptake is responsible for atypical meningioma growth. Derangement of mitotic spindles may lead to polyploid chromosome sets and thereby to giant nuclei. This seems likely because induction of tumor growth after iron injection has been described. Spodographic examinations showed that even histologically inconspicuous tumor cells contain fine-grained iron. Application of the Prussian-blue reaction to spodograms reveals distinctly higher iron contents than common paraffin sections had proved.

Cell Nucleus↗
PubMed1992

Macrophages in normal human bone marrow and in chronic myeloproliferative disorders: an immunohistochemical and morphometric study by a new monoclonal antibody (PG-M1) on trephine biopsies.

An immunohistochemical and morphometric study was performed on routinely processed trephine biopsies of the bone marrow in 30 normal individuals and in 90 patients with various subtypes of chronic myeloproliferative disorder. Using a new monoclonal antibody (PG-M1) directed against a formalin-resistant epitope on macrophages and by employment of the Prussian blue reaction, quantitation of this cell population was feasible. Morphometric analysis revealed that the number of iron-laden macrophages represented only a fraction of the total number of histiocytic reticular cells. As could be expected, in polycythaemia rubra vera, no haemosiderin deposits were detectable, but the content of macrophages slightly exceeded that of the normal bone marrow. In chronic myeloid leukaemia 9 of 30 patients showed a significant increase in PG-M1-positive reticular cell elements. These were consistent with pseudo-Gaucher cells, sea-blue histiocytes and intermediate cell types. Primary (idiopathic) myelofibrosis-osteomyelosclerosis was characterized by a significant increase in macrophages (25 of 30 patients). Involvement of macrophages in the complex mechanisms generating bone marrow fibrosis and angiogenesis and in bone remodelling (osteosclerosis) may be responsible for this finding.

Adipose Tissue↗
PubMed1976

A glass-insulated "Elgiloy" microelectrode for recording unit activity in chronic monkey experiments.

A glass-coated microelectrode made of a stiff cobalt--nickel alloy is described. The thickness of the glass coating and the length of the uninsulated tip can be varied as desired. The electrode is stiff enough to insert through the relatively tough dura of the monkey in chronic experiments. Further, the recording site can be marked by electrolytic deposition of iron and the Prussian blue reaction.

Alloys↗