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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↗
PubMed1998

Colloidal iron staining in renal epithelial neoplasms, including chromophobe renal cell carcinoma: emphasis on technique and patterns of staining.

Positive staining with Hale's colloidal iron stain, or modifications thereof, is considered a diagnostic feature for chromophobe renal cell carcinoma and has been used as a discriminatory feature to differentiate it from other renal tumors. We studied colloidal iron staining in 62 cases encompassing a wide histologic spectrum of renal neoplasms (14 chromophobe renal cell carcinomas, 19 renal oncocytomas, 11 each of granular variants and conventional clear cell renal cell carcinomas, and 7 eosinophilic variants of papillary renal cell carcinoma) to investigate the specificity of the stain for chromophobe renal cell carcinoma. In cases of chromophobe renal cell carcinoma, sections from two different areas were stained to ascertain whether there was any spatial variation in staining. Influence of staining techniques on the results also was investigated by staining each case of chromophobe renal cell carcinoma using two different methods: the traditional Hale's and a modified Mowry's technique, which treats sections with 3% acetic acid before adding the colloidal iron. Our results show that positive staining with colloidal iron stain is not limited to chromophobe renal cell carcinoma, however, a diffuse and strong, reticular staining pattern was observed only in cases of chromophobe renal cell carcinoma (14 of 14). The staining patterns were less consistent in all other renal neoplasms and differed from the reaction observed in chromophobe renal cell carcinoma. Most renal oncocytomas (16 of 19) had focal and weak, fine dustlike positivity, and all clear cell carcinomas showed focal, coarse, dropletlike positive staining (22 of 22), in addition to a focal, coarse, bubbly pattern in 5 of 11 clear cell subtypes. Although all seven cases of the eosinophilic variant of papillary renal cell carcinoma showed strong, coarse, dropletlike staining, most of the positivity was coincident with the Perl's (prussian blue) reaction, indicating that the staining was due to hemosiderin, which is frequently present in this histologic subtype of renal cell carcinoma. Staining intensity did not vary considerably among different areas of chromophobe renal cell carcinoma, but the modified Mowry's method yielded brighter and sharper reticular staining, as compared with the Hale's method. Our results show that in the appropriate morphologic context diffuse and strong reticular positivity using the modified Mowry's colloidal iron stain method is highly characteristic for chromophobe renal cell carcinoma. Treatment of sections with 3% acetic acid before adding the colloidal iron gives technically superior staining results.

Adenocarcinoma, Clear Cell↗
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↗
PubMed1996

Quantitative photometrical assessment of iron deposits in synovial membranes in different joint diseases.

We investigated 86 synovial membranes from patients suffering either from rheumatoid arthritis (RA) or osteoarthritis (OA). Iron deposits in the synovial membrane were stained by the Prussian blue reaction, and the amount of stained iron was quantitatively assessed by microscope photometry. We found a statistically significant increase in iron deposits in the synovial membrane of RA patients when compared to OA patients. The amount of iron deposits correlated with the histological subtype of synovitis, those presenting with more exudative and proliferative features showing greater amounts of iron deposits. We also observed an inverse correlation between the haemoglobin concentration and erythrocytes in the serum and the amount of iron in the synovial membrane. From our data we concluded that iron deposits in the synovial membrane can contribute by several mechanisms, including activation of oxygen radicals, to the chronic inflammatory reaction in RA synovitis.

Aged↗
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↗
PubMed1986

Median eminence equivalence of the neurohypophysis of the hagfish, Eptatretus burgeri.

Three substances of different molecular size were injected into the ventricular space (III) of the neurohypophysis of anesthetized hagfish, Eptatretus burgeri, to learn whether such substances could diffuse across the connective tissue barrier between the neuro- and adenohypophysis. The substances tested were trypan blue, a vital dye that is a fine particulate colloid; horseradish peroxidase (HRP), an enzyme protein whose distribution in tissue can be revealed through its chromogenic action on a substrate; and ferric ion, whose distribution can be revealed through the Prussian blue reaction. All three substances were transferred across the neurohypophyseal wall to the extracerebral area within a few minutes, the trypan blue and HRP apparently through action by tanycytes. The trypan blue, within a 15-min period, moved no further. The HRP protein diffused through connective tissue as far as the dorsal (2 min) and ventral (5 min) borders of the adenohypophysis, but did not enter the secretory cells themselves. The ferric ion reached the adenohypophysis quickly (2 min) and entered the secretory cells in higher concentration than it was in the surrounding connective tissue. These data indicate that the hagfish ventral neurohypophysis is functionally capable of supplying neurosecretory regulatory factors to the adenohypophysis. It is thus a "diffusional median eminence." We can propose that the cyclostome median eminence, lacking a portal relation to the adenohypophysis, represents the phylogenetically primitive brain--pituitary relationship.

Animals↗
PubMed2003

Coeruleotrigeminal inhibition of nociceptive processing in the rat trigeminal subnucleus caudalis.

It has been accepted that the descending system from the nucleus locus coeruleus (LC)/nucleus subcoeruleus (SC) plays a significant role in spinal nociceptive processing. The present study was designed to examine modulation of nociceptive processing in the caudal part of the trigeminal sensory nuclear complex, the trigeminal subnucleus caudalis which is generally considered to be involved in the relay of oral-facial nociceptive information. Experiments were performed on anesthetized Sprague-Dawley rats. The site of LC/SC stimulation was confirmed by histology using potassium ferrocyanide to produce a Prussian blue reaction product marking the iron deposited from the stimulating electrode tip. Only data from rats which had electrode placements in the LC/SC were used. Electrical stimulation was delivered at a stimulus intensity below 100 microA in the present study. Stimulation at sites inside the LC/SC produced a reduction of both spontaneous activity and responses of subnucleus caudalis neurons to somatic input, especially nociceptive input. Increasing stimulation frequency in the LC/SC resulted in an increase in inhibitory effects on nociceptive responses of subnucleus caudalis neurons. At three of nine sites outside the LC/SC, electrical stimulation was effective on descending inhibition. A significant difference in the inhibitory effects was observed when the inhibitory effects were compared between sites of stimulation inside the LC/SC and three effective sites of stimulation outside the LC/SC. These findings suggest that nociceptive processing in the subnucleus caudalis is under the control of the descending modulation system from the LC/SC. To understand the effects of repetitive stimulation with high frequency on fine unmyelinated LC/SC fibers, the existence of recurrent collateral excitation in the LC/SC may be considered.

Action Potentials↗
PubMedDate not supplied

Macroscopic and microscopic aspects of collared peccary and white-lipped peccary placenta.

This study examines middle and late gestational placentae from 13 Tayassu tajacu (collared peccary) and 3 Tayassu pecari (white-lipped peccary), which are Artiodactyla belonging to the Family Tayassuidae. The chorionic sac of Tayassu species is diffuse and chorioallantoic. These epitheliochorial placentae show no trophoblast invasion into the uterine epithelium and there is interdigitation between fetal and maternal microvilli. Two distinct regions of the fetomaternal interface can be identified: the interareolar and the areolar regions. The uterine epithelium has eosinophilic cytoplasm with dispersed, basophilic and electron-dense granules. Trophoblast cells are irregularly cuboidal on top of the fetal ridges and columnar on troughs, where cells have cytoplasmic vesicles and large basal vacuoles, surrounded by whorls of smooth membranes. Capillaries indent the trophoblast cells forming a placental barrier 3 microm or less thick. The columnar uterine glandular epithelium has a subpopulation of granules staining with Perl's Prussian blue reaction, suggesting iron secretion. In areolar areas, the trophoblast cells show apical microvilli, a basophilic cytoplasm with electron-dense intracellular vacuoles and cisternae. The placenta can therefore be classified as non-deciduate. The ultrastructural aspects of this study reveal features that have not previously been described and extend our knowledge of functions relating to materno-fetal transport in these species.

Animals↗
PubMed2001

Trigeminally innervated iron-containing structures in the beak of homing pigeons, and other birds.

The ophthalmic nerve in the upper beak was labelled with cholera toxin B-chain, and iron was identified using the Prussian Blue reaction. Iron deposits were found in the caudal part of the beak, and some were concentrated in cells that clustered in encapsulated structures densely innervated by ophthalmic nerve fibres. Such structures could form the anatomical basis of a type of mechanoreceptor that transmits magnetic sense information to the brain.

Animals↗
PubMed1994

Increased tissue endothelin immunoreactivity in atherosclerotic lesions associated with acute coronary syndromes.

Acute coronary syndromes are accompanied by exaggerated vasoconstriction. Since thrombin induces production of the potent vasoconstrictor endothelin-1, we used immunohistochemistry on coronary atherosclerotic specimens from 30 consecutive patients. Endothelin-1-like immunoreactivity was present in 5 of 9 (55%) lesions from patients with stable angina, in 6 of 7 (86%) from crescendo angina, and in all 14 from angina at rest. Endothelin-1 immunoreactivity was most common in areas with a positive Prussian-blue reaction indicative of previous intraplaque haemorrhage. Tissue endothelin-1-like immunoreactivity might contribute to the exaggerated coronary vasoconstriction.

Acute Disease↗
PubMed1998

In vivo microelectrode track reconstruction using magnetic resonance imaging.

To obtain more precise anatomical information about cortical sites of microelectrode recording and microstimulation experiments in alert animals, we have developed a non-invasive, magnetic resonance imaging (MRI) technique for reconstructing microelectrode tracks. We made microelectrode penetrations in the brains of anesthetized rats and marked sites along them by depositing metal, presumably iron, with anodic monophasic or biphasic current from the tip of a stainless steel microelectrode. The metal deposits were clearly visible in the living animal as approximately 200 microm wide hypointense punctate marks using gradient echo sequences in a 4.7T MRI scanner. We confirmed the MRI findings by comparing them directly to the postmortem histology in which the iron in the deposits could be rendered visible with a Prussian blue reaction. MRI-visible marks could be created using currents as low as 1 microA (anodic) for 5 s, and they remained stable in the brains of living rats for up to nine months. We were able to make marks using either direct current or biphasic current pulses. Biphasic pulses caused less tissue damage and were similar to those used by many laboratories for functional microstimulation studies in the brains of alert monkeys.

Animals↗