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PubMed1975

[Histomorphological and spectroscopic studies on the skeleton following alloplastic joint substitution (author's transl)].

We report on our morphological and spectrometric findings with complete acetabuloplastics. The histomorphological investigation revealed the formation of a narrow cicatricial coat round the prothesis. The collagenous tissue borders on a zone of granulation tissue of varying width. Granular foreign matter can be found in cicatrical tissue especially in the extracellular space and in granulation tissue in macrophages. Only a few partigles turned out to be iron in the Prussian blue reaction. Spectral analysis proved the hip and thigh-bones to contain more than 0.001 p. c. of cobalt and chrome. Toxicological problems inherent in the use of the cobalt alloy "Vitallium" are discussed. The dangers of metal poisoning are dealt with in addition to those of the development of osteomyelitis and the formation of a sarcoma. At last we discuss our macroscopic findings concerning the fit of the prosthesis, the metal demonstration by electron micrsocopy, and the scanning electron microscopic investigation of the metal surfaces.

Chromium↗
PubMed1975

Permeability of the post-ischemic rat aorta.

The time course of the vascular permeability disorder following upon acute hypoxia and the fate of plasma substances entering the vessel wall in consequence of increased permeability were studied on the rat abdominal aorta rendered hypoxic for one hour by double ligature and recirculated for periods ranging from one hour to 30 days. The distribution and quantity of mural plasma inhibition were determined histochemically by means of a colloidal iron tracer and Prussian blue reaction and by photometric analysis, respectively. Plasma imbibition reached its maximum after recirculation for 24 to 48 hours and fell to an almost normal level after 10 days. Administration of the colloidal iron tracer on the second day of recirculation, when the permeability disorder was at its peak, showed plasma imbibition in every layer of the vessel wall. At seven days it was restricted to the outer third of the media and the adventitia. The endothelium is acting as the main barrier to mural plasma imbibition and in the case of a permeability disorder only the elastic lamellae constitute a temporary mechanical obstacle to the ingression of plasma.

Animals↗
PubMed1990

Evidence of transferrin binding sites on the surface of Leishmania promastigotes.

A glycoprotein of 78,000 molecular mass (78 kDa), associated with the membrane of Leishmania infantum promastigotes, was identified and immunopurified by monoclonal antibody (mAb) LD9 produced against isolated membrane preparations. mAb LD9 was subsequently found to bind to human transferrin, also of 78 kDa. Binding of LD9 to transferrin was completely abolished when the mAb was preabsorbed by Leishmania membranes, thereby indicating that the 78-kDa Leishmania membrane-associated glycoprotein and transferrin have common antigenic epitope(s). The 78-kDa Leishmania membrane-associated protein was released in soluble nonaggregated form by mild treatment with acetic acid saline. Anti-transferrin polyclonal antibodies, recognized both the membrane-associated and the soluble form of the 78-kDa glycoprotein. The 78-kDa soluble form was characterized further as an iron-containing protein. The above data combined with iron uptake by promastigotes as demonstrated by the Prussian blue reaction indicate that the 78-kDa Leishmania membrane-associated glycoprotein is transferrin. The binding of 125I-human transferrin to Leishmania-purified membrane preparations was then investigated. The results indicate the presence of a high affinity saturable binding site (Kd = 2.2 10(-8) M) that is specific for transferrin. We suggest that the 78-kDa glycoprotein recognized by mAb LD9 is transferrin that binds to the surface of Leishmania promastigotes via a transferrin receptor.

Animals↗
PubMed1990

[Disturbances in iron utilization in acute leukemia and preleukemia].

With Prussian blue reaction nonhaemoglobin iron in the erythroblasts is demonstrable. Three pathological sideroblast types are recorded separately: abnormal intermediate type I and II sideroblasts and ring sideroblasts, representing increasing levels of sideroachrestic disturbance. This permits the classification of sideroachrestic disturbances into four degrees of seriousness. The frequency of a sideroachrestic disturbance in 47 untreated patients with acute myeloid leukaemia was 87%. Among 11 patients with preleukaemic condition, 8 had a disturbance of iron utilisation. In both preleukaemia and leukaemia mainly intermediate sideroblasts were present. All patients with preleukaemia developed leukaemia within 1-20 months. In the course of preleukaemic condition a slight increase of iron misutilisation was obvious when terminating in overt leukaemia. This could be of prognostic importance. After treatment, pathological sideroblasts disappeared only in 2 out of 15 patients with complete remission. There was no correlation between effect of therapy and course of iron misutilisation.

Acute Disease↗
PubMed1977

[Iron containing nuclear inclusions in human pigmentary cirrhosis (author's transl)].

In human pigmentary cirrhosis nuclear (pseudo-)inclusions of cytoplasmic material, containing less or more degenerated and therefore faintly stained hemosiderin granules, are to be observed. But sometimes there are also finely fibrillar or granular proteinaceous materials, stainable by the Prussian-blue reaction, lying between the chromatin-strands or occupying the whole nucleus and displacing the chromatin to the nuclear envelope (margination of chromatin). Such uncoloured substances may condense into homogeneous masses and nearly hexagonal (0r related) crystals with a diameter up to 14 micron and a yellow-brownish colour, giving a strongly positive PERL's reaction. In contrast to the preceding stages intranuclear crystals of this kind have been observed in one case only. After destruction of the nuclear envelope and the marginated chromatin the crystals are lying free in the cytoplasm and later on, the cytoplasm being destroyed too, they may be ingested by von Kupffer cells. All the iron containing crystals, to be found in the cytoplasm, derive from former intranuclear inclusions. The intranuclear deposits of iron containing protein are interpreted as ferritin-aggregates. It is supposed that ferritin molecules, built up in the cytoplasm, do enter the nucleus via the pores of the nuclear envelope. Such an event not only signalizes a cytopathologic reaction but in turn may give rise to such additional cytopathologic lesions as cell shrinking and cell death.

Aged↗
PubMed1978

Pituitary siderosis. A histologic, immunocytologic, and ultrastructural study.

Pituitaries of 6 patients with various iron overload states have been studied by morphologic techniques, including immunostaining and electron microscopy. The immunoperoxidase technique combined with the prussian blue reaction revealed iron deposition in all five adenohypophysial cell types, indicating the iron uptake per se does not entirely block hormone storage. Iron distribution was uneven; more iron was demonstrated in PAS-positive cells than in orangeophils, and a preferential localization was disclosed in the gonadotrophs. In 2 cases of hemochromatosis, reduction of pituitary gonadotrophs was implicated in the genesis of hypogonadism. By electron microscopy, iron particles were noted in the cytoplasm of various adenohypophysiocytes, partly incorporated into lysosomes. Some adenohypophysiocytes with iron accumulation showed degranulation by light and electron microscopy and decreased hormone storage by the immunoperoxidase technique. Although these changes may be causally related to iron deposition, more work is required to prove that iron has a direct toxic effect on adenohypophysial cells.

Adult↗
PubMed1976

Alterations of the myoid pericanalicular layer in liver. A light microscopical pilot study of human autopsy material.

Previous histochemical studies demonstrated a myoid layer along biliary pathways. This report describes alterations of the pericanalicular layer in various lesions. Carnoy- or methacarn-fixed sections of livers from 200 autopsies were treated with the tannic acid-phosphomolybdic acid-dye method for myosins. In other series, methods for myosins were combined with the picro-Sirius Red F3BA stain or the Prussian Blue reaction. In normal liver, bile canaliculi were lined by a narrow myoid layer. In other cases, eg, hepatic cirrhosis, the pericanalicular layer showed slight to moderate thickening. In cases with obstruction or atresia of bile ducts, the dilated canaliculi were surrounded by a prominent myoid layer. These observations support theories that the pericanalicular layer is contractile and promotes bile flow under positive pressure.

Bile Ducts, Intrahepatic↗
PubMed1977

Marrow iron.

Marrow aspirates stained with the Prussian blue reaction give a variety of patterns. Characteristic pictures are shown for megaloblastic anemia, infection, and subjects after injection of iron dextran. The appearance of iron in histiocytes in marrows stained with Wright's stain is differentiated from noniron deposits. A graph shows the relative distribution of iron in RBCs and storage in different clinical situations.

Anemia↗
PubMed1996

Pigment in prostatic epithelium and adenocarcinoma: a potential source of diagnostic confusion with seminal vesicular epithelium.

Granular cytoplasmic pigment is frequently observed in prostatic epithelium, and may cause confusion with ejaculatory ductal and seminal vesicular epithelium. We evaluated the distribution and histochemical and immunohistochemical phenotype of pigment in nine serially sectioned, whole-mounted prostatectomy specimens with carcinoma. Golden yellow-brown pigment was found in prostatic glandular epithelium in eight cases (89%) of nine, varying from inconspicuous and focal to extensive and diffuse. It was usually located in the basal portion of the secretory (lumenal) cell layer, but was also observed throughout the cytoplasm, and rarely seen in basal cells. Stromal pigment was observed in seven cases (78%) of nine and was patchy and inconspicuous. Epithelial pigment was observed throughout the prostate (transition zone, 67%; central zone, 56%; peripheral zone, 89%; and periurethral glands, 56%). Focal pigment was observed in high-grade prostatic intraepithelial neoplasia in seven cases (78%) of nine and one case (11%) of nine cases of adenocarcinoma. The epithelial pigment was histochemically similar to lipofuscin pigment (Fontana-Masson positive, bleached by permanganate reaction, Prussian blue negative, and Ziehl-Neelsen positive); S100 protein immunohistochemical stains were negative. Our results indicate that pigment is frequently present in the prostatic epithelium of all zones, at least focally, and has histochemical similarities to pigment in the seminal vesicular epithelium. Awareness of prostatic epithelial pigment is important, and the differential diagnosis of pigmented epithelium in scant specimens such as those obtained from needle biopsies includes normal and neoplastic prostatic epithelium, seminal vesicle epithelium, and ejaculatory duct epithelium.

Adenocarcinoma↗
PubMed1993

A sensitive post-DAB enhancement technique for demonstration of iron in the central nervous system.

A technique is described for enhancing the reaction product of the staining reaction for iron in paraffin-embedded tissue from central nervous system (CNS). After amplification of the Prussian Blue staining reaction with 3,3'-diaminobenzidine (DAB), the reaction product was further intensified using a stepwise treatment with silver methenamine, gold chloride and uranyl nitrate (post-DAB treatment). Following the Prussian Blue-DAB staining reaction, iron was seen only in glial cells and choroid plexus epithelial cells, whereas the post-DAB treatment revealed that neurons and endothelial cells of the brain capillaries were also positively stained. The post-DAB treatment resulted additionally in an increased intensity of the reaction product within choroid plexus epithelial cells compared to that obtained in sections subjected only to the Prussian Blue-DAB reaction. The reliability of the method was evaluated using liver sections as positive controls. Furthermore the higher sensitivity of the method was assessed using nitrocellulose filters containing serially diluted iron-saturated transferrin. The post-DAB method is simple and can easily be applied to formalin- or glutaraldehyde fixed, paraffin-embedded nervous and non-nervous tissue.

3,3'-Diaminobenzidine↗
PubMed1995

Identification of Fourier transform infrared signals from the non-heme iron in photosystem II.

Fourier transfer infrared (FTIR) signals originating from a high-potential electron acceptor in PS II were studied by flash-induced FTIR difference spectroscopy. Redox titration of these signals using ferri-ferrocyanide mixtures showed midpoint potentials (Em) of 489 +/- 12 and 426 +/- 9 mV at pH 5.5 and 6.5, respectively, revealing a pH dependence of about -60 mV/pH unit. These Em values and pH dependence were in good agreement with those of the non-heme iron, so-called Q400, located between QA and QB. This indicates that the observed FTIR signals are due to changes in ligands of the non-heme iron and surrounding protein moieties induced on photoreduction from Fe3+ to Fe2+.

Electron Transport↗
PubMed1978

[Ultrastructural localization of 3-beta-hydroxysteroid dehydrogenase activity in the glandular tissue cells of the testis of Pleurodeles waltlii Michach. (Amphibia, Urodela)].

With the potassium ferricyanide method, the localization of the sites of 3 beta-hydroxysteroid dehydrogenase activity has been investigated in the glandular tissue cells, previously fixed, of the Pleurodeles testis. An abundant copper ferrocyanide precipitate, the final reaction product, is observed in the vicinity of the external faces of the membranes of the smooth endoplasmic reticulum, or in contact with them. A very weak reaction occurs in mitochondrial cristae.

3-Hydroxysteroid Dehydrogenases↗
PubMed2003

[Demonstration of trivalent iron in decalcified bone marrow specimens. Comparative study of standard Perls stain on sections and the Perls pre-reaction on fragments].

UNLABELLED: In the literature the usefulness of the Perls reaction for Iron (Fe+++) stores estimation on bone marrow is largely accepted. However, due to decalcification, there is a considerable disagreement about the accuracy of this staining on osteomedullary tissue. In fact loss of Fe+++ has been claimed by some Authors, and denied by others. In order to check this controversial point in this paper Perls reaction, as a methodological variant, was directly anticipated on fragments (pre-Perls). In this way numerous osteomedullary sternal specimens from autopsies (non hematological cases) were observed in comparison with normal Perls reaction on sections (standard-Perls). In standard-Perls series (A) decalcification was achieved by HCl (alone or in combination with EDTA) or by formic acid. On pre-Perls treated fragments decalcification was an intrinsic effect of the hydrochloride component during Prussian Blue formulation. Nine different procedures of fixation were tested respectively for standard-Perls and pre-Perls. Pre-Perls was performed on pre-fixed (B series) as well as post-fixed material (C series). For the microscopical evaluation was applied the classical grading system proposed by Lundin for histology. Maximum grade was only observed in B and C, was the same in both these series and varied from 2 to 4 depending on autopsic case. Results strongly indicate the superiority of pre-Perls versus standard-Perls, with a differential grading ranging from 2 to 4 grades, in seven procedures of B (two procedures were equivalent in A and B) and in all the procedures of C. The comparison between B and C series showed inferiority of B in four procedures. For control (D series) standard-Perls was systematically performed on sections from pre-Perls: no differences between pre-Perls and pre-Perls + standard-Perls were observed indicating a complete permeation of fragments by Perls reagents. In general terms morphology in pre-Perls B was better than in pre-Perls C and substantially identical to standard procedures of A. Other data are analytically reported. CONCLUSIONS: Pre-Perls staining (with special reference to five procedures of B) seems to be recommendable for decalcified bone marrow biopsies, in conjunction with standard processing, when an high sensitivity in detection of Fe+++ deposits is required or in absence of aspirate. Pathologists could subsequently decide if this methodological variant is suitable or not for an effective alternative choice in their routine according to morphological and immunohistochemical adequacy.

Bone Marrow↗
PubMed1982

Iron deposits surrounding multiple sclerosis plaques.

Autopsy samples from cerebral areas of five brains from patients with multiple sclerosis (MS) and from six control brains were stained with Perls acid ferrocyanide to detect nonheme iron present as hemosiderin. Positive iron reactions were observed only in MS sections surrounding demyelinated plaques. Myelinated white matter near the lesion contained numerous iron-laden ovoid bodied and axons that stained positively for iron. Positive reactions were also found within blood vessels of gray matter near the lesion. A possible source of the iron was extravasated blood.

Adult↗
PubMed2006

Phenotypic study of human gingival fibroblasts labeled with superparamagnetic anionic nanoparticles.

BACKGROUND: A specific labeler of the human gingival fibroblast (HGF) does not exist. Anionic maghemite nanoparticles allow labeling of a wide cell variety and their recognition in cellular, organotypical, and animal models. METHODS: We studied internalization effects of nanoparticles on an HGF phenotype in vitro, evaluating transcription and secretion of connective tissue remodeling molecules, i.e., matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and cytokines controlling their activation/inhibition, i.e., transforming growth factor-beta (TGF-beta1), tumor necrosis factor-alpha (TNF-alpha), and interleukins 1beta and 4 (IL-1beta and IL-4). After proliferation kinetics, cellular uptake was studied by Perls coloration and magnetophoresis on labeled culture. Dot blotting, Western blotting, and zymography were used to detect MMP-1, -2, and -3 and TIMP-1 and -2 secretions in culture supernatants, and reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression of these molecules. Enzyme-linked immunosorbent assay (ELISA) tests were used to determine TGF-beta1, TNF-alpha, IL-1beta, and IL-4 levels. RESULTS: Our data indicated high (15.3+/-5.8 pg/cell) but heterogeneous distribution of nanoparticles in HGF. Twenty-four hours after labeling, MMP-1, -2, and -3 and TIMP-2 secretion increased (P<0.001) with RT-PCR confirmation at 12 hours, whereas TIMP-1 did not. IL-1beta increased at day 1 (D1) (P<0.001) and IL-4 at D3 (P<0.01), but not TGF-beta1 or TNF-alpha. CONCLUSIONS: After labeling with these maghemite nanoparticles, HGF increased secretion of IL-1beta at D1, probably inducing the increase of MMP-1, -2, and -3 and TIMP-2. The increase of IL-4 secretion began with the decreased synthesis of MMPs and TIMPs at D3. Despite this transitory inflammatory reaction at 3 days following internalization, maghemite nanoparticles did not affect HGF phenotype, thereby authorizing their use as labelers.

Adult↗
PubMed1984

Iron deposition in the integument of lampreys.

The integument of larval, parasitic adult, and upstream-migrant lampreys (Petromyzon marinus) was examined for iron deposition using light microscopic histochemistry and routine and histochemical procedures in the electron microscope. Ferritin particles, representing ferric iron, are present throughout most of the cytoplasmic matrix and within dense granules and vacuoles of epidermal mucous cells, but are not located in skein or granular cells. These particles are abundant in mucous cells of the dorsal surface but not the ventral surface and are more concentrated in adult lampreys compared to larva. Histochemistry revealed only sparse amounts of ferrous iron. Iron is not present in the dermis but is found in adipocytes of a subcutaneous layer. The deposition of integumentary iron is discussed with reference to body pigmentation and excretion of this metal.

Adipose Tissue↗
PubMed2003

Ultrastructural analysis of a putative magnetoreceptor in the beak of homing pigeons.

With the use of different light and electron microscopic methods, we investigated the subcellular organization of afferent trigeminal terminals in the upper beak of the homing pigeon, Columba livia, which are about 5 microm in diameter and contain superparamagnetic magnetite (SPM) crystals. The SPM nanocrystals are assembled in clusters (diameter, approximately 1-2 microm). About 10 to 15 of these clusters occur inside one nerve terminal, arranged along the cell membrane. Each SPM cluster is embedded in a solid fibrous cup, open towards the cell surface, to which the cluster adheres by delicate fiber strands. In addition to the SPM clusters, a second inorganic iron compound has been identified: noncrystalline platelets of iron phosphate (about 500 nm wide and long and maximally 100 nm thick) that occur along a fibrous core of the terminal. The anatomic features suggested that these nerve endings could detect small intensity changes of the geomagnetic field. Such stimuli can induce deformations of the SPM clusters, which could be transduced into primary receptor potentials by mechanosensitive membrane receptor channels. The subepidermal fat cells surrounding the nerve endings prevent the inside from external mechanical stimuli. These structural findings corresponded to conclusions inferred from rock magnetic measurements, theoretical calculations, model experiments, and behavioral data, which also matched previous electrophysiologic recordings from migratory birds.

Animals↗
PubMed1985

Sequential histochemical staining for resident and recruited macrophages.

A new histochemical technique is described that permits differentiation of resident from recruited macrophages by staining of paraffin sections of tissues from rats and mice. Resident macrophages are identified by their ability to phagocytose and retain intravenously injected colloidal Prussian blue. New macrophages that emigrate into tissue are identified by phagocytosis of a second colloid, iron dextran. Paraffin sections of formalin-fixed tissues are sequentially stained for the presence of the two colloids with different chromogens, the endogenous pseudo-peroxidase activity of colloidal Prussian blue used to catalyze the polymerization of diaminobenzidine and after conversion of iron dextran to Prussian blue, the second colloid used to catalyze the polymerization of tetramethylbenzidine. The staining results in resident macrophages staining brown while newly recruited macrophages stain blue. The studies have shown that colloidal Prussian blue is stable in vivo and neither loses its catalytic activity nor undergoes extensive redistribution. They also show that the technique can be used to measure Kupffer cell recruitment stimulated by complete Freund's adjuvant in rats and tumor-associated macrophage recruitment in subcutaneous and spontaneous liver metastases in mice.

Animals↗