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[A case of primary amyloidosis associated with giant cell infiltration within a Bowman's capsule].

A 67-year-old man was hospitalized with a diagnosis of nephrotic syndrome. Physical findings at admission were generalized edema and macroglossia. Urinalysis showed massive proteinuria, + +occult blood, and granular and broad casts. Ig A lambda monoclonal gammopathy was noted in the serum. There was no evidence of myeloma in the bone marrow aspirate, scintigram or X-ray of the bone. A biopsy specimen of the kidney showed massive deposits of structureless material in the glomeruli. Marked cell infiltration was also observed in the interstitium. Multinucleated giant cells were occasionally seen in the Bowman's capsules and the interstitium. There were reactive changes in the Bowman's capsule adjacent to the giant cell. The deposits were proved to be amyloid by positive staining with Congo red and apple-green birefringence by polarized light. In addition, microfibrills seen on electron microscopy displayed deposits. Amyloid depositions were observed in other tissues such as gingiva, skin and tongue. Staining of amyloid with Congo red was resistant to potassium permanganate, and amyloid was positively stained with lambda-light chain of immunoglobulin. These findings indicated that the patient had primary amyloidosis. Infiltration of the multinucleated giant cell has been reported only in patients with familial amyloidosis and secondary amyloidosis associated with rheumatoid arthritis. To our knowledge the present case is a first report of the giant cell infiltration in a Bowman's capsule in primary amyloidosis.

Aged↗

Ultrastructural immunohistochemical localization of polyclonal IgG, C3, and amyloid P component on the congo red-negative amyloid-like fibrils of fibrillary glomerulopathy.

Renal biopsies from seven patients with Congo red-negative amyloid-like fibrillary glomerulopathy (FGP) were examined by protein A gold immuno-electron microscopy. Ultrastructurally, the fibrils in all cases exhibited positive immunostaining for IgG, both Ig light chains, C3, and amyloid P component (AP), but did not show positive immunostaining for glomerular basement membrane (GBM)-associated proteins (collagen type IV and heparan-sulfate proteoglycans) or microfibril-associated proteins (fibronectin and fibrillin). In a triple-label study, AP and IgG were colocalized along the same fibril, whereas the gold probes for the detection of collagen type IV were absent. The results suggest that the fibrils are comprised of polyclonal IgG and C3 that bind AP. AP was immunolocalized sparsely but regularly along the lamina rara interna of normal GBM. AP was absent in the fibrils in a case of diabetic glomerulopathy, was scattered randomly without specificity for the electron-dense deposits in the GBM of membranous glomerulopathy, and lined up regularly along the fibrils in amyloid deposits. FGP is an entity in which the fibrils bind AP but lack the beta-pleated sheet structure necessary for Congo red staining that is typical of amyloid.

Amyloid↗

[Formation of the peritrophic membrane in the gut of Colicoides punctatus (Diptera: Ceratopogonidae)].

Using light and electron microscopy, the structure of the peritrophic membrane (PM) was studied in females of the biting midge Culicoides punctatus (Mg.) during the process of blood meal digestion. The PM formation occurs in the posterior part of midgut and lasts during the most time of the digestive cycle. The PM precursors are probably not associated with any intracellular granules. The PM consists of two main components: light structural component and dark amorphous matrix, both of which are directly released from the entire microvillar surface. The aggregation of secreted components takes place in the gut lumen to form gel-like multilayered PM up to 6 microns thickness with bundles of microfibrills situated in the PM surface facing the lumen. Similarities and differences of the PM formation in most groups of blood-sucking insects are discussed.

Animals↗

A cell line derived from BBN (N-butyl-N-[4-hydroxybutyl]-nitrosamine)-induced rat bladder cancer: establishment and scanning electron microscopic cell surface characteristics.

This research was performed to establish a cell line from experimental bladder tumor and to discuss the biological characteristics of the cell line so established. Tissue cultures of epithelial cells were derived from a rat bladder cancer induced by BBN. The cells showed loss of contact inhibition and the phenomenon of piling up after several subcultures. Colonial cloning was used. The population doubling time of the wild strain and the colonial clones was about 30 h. The chromosomal mode ranged from triploid to tetraploid to tetraploid. Plating efficiency was well below 20%. Intraperitoneal backtransplantation into newborn Wister rats resulted in tumors in all cases. These tumors, in some parts, resembled primary transitional cell carcinoma. The major tumor cell groups, however, showed marked keratinization and the picture of squamous cell carcinoma. The nucleus/cytoplasm ratio and the numbers of nuclei, free ribosomes and intracytoplasmic microfibrils were increased. Dense microvillus arrangements characterized the electron microscopic picture. During the mitotic phase, the cells became large and globular whereas the microvilli were relatively short and were gathered profusely over the whole surface. Cells in the gap 1-synthetic phase developed lamellipodia and pseudpodia-like cytoplasmic processes and were polygonal in shape. Microvilli were present in the central part containing the nucleus, but their numbers were somewhat decreased and their height increased (scanning electron microscopy).

Animals↗

[Cardioprotective effect of energostim in toxic allergic myocarditis].

Energostim is a combined drug comprising a mixture of nicotinamide adenine dinucleotide (0.5 mg), cytochrome C (10 mg), and inosine (80 mg), representing antihypoxant and antioxidant of direct action in one ampule. After pretreatment and subsequent 3-day energostim therapy of animals with 3-day toxico-allergic myocarditis (3d-TAM), the ECG was free of any rhythm disorders and showed evidence of improved conduction, restoration of the normal form of T-wave and the position of ST segment, while the content of myofibrillar fraction of creatine phosphokinase and toxic products of disturbed metabolism (degree of endotoxemia) decreased to the upper normal level. Under the action of energostim, neither pressure nor the maximum rate of pressure buildup in the left ventricle are reduced (as they do upon 3d-TAM); neither systolic and diastolic functions are disturbed, nor their coordination (r = 0.79 between dP/dtmin and dP/dtmax, p < 0.01). The restoration of contractile activity and maximum rate of relaxation of myocardial microfibrils during 3d-TAM is accompanied by an increase in the content of adenyl nucleotides, in the ATP/ADP, ADP/AMP, NAD/NADH, and NADP/NADPH ratios, and in the cytosol phosphorylation potential. The energostim-induced improvement in the energy supply system are accompanied by restoration of the ability of sarcoplasmic reticulum to efflux Ca2+. Thus, it is demonstrated that the effect of energostim is related to its ability to actively participate in intracell metabolic processes in myocardium, abolish necrotic changes and endotoxicosis, and restore homeostasis in the systems responsible for the contraction--relaxation process (thus preventing from the development of dysfunction of the left ventricle and the heart failure).

Animals↗

Clinical significance of elastin turnover--focus on diseases affecting elastic fibres.

Elastic fibres are an important component of connective tissue. They are composed of two elements: protein-elastin and microfibrils. Tissue rich in elastin include: aorta and major vessels, lungs, ligaments, tendons and skin. Elastases are a group of enzymes, which are able to degrade elastin. They are liberated from different cells including granulocytes, monocytes, lymphocytes, skin fibroblasts, cancer cells and others. The product of elastin degradation is known as elastin-derived peptides (EDP) and is a marker of elastolysis. Elastin-derived peptides are thought to act on elastin-laminin receptor (ELR), which is located on membrane of fibroblasts, granulocytes, lymphocytes, monocytes and' cancer cells. An increased elastin turnover can occur in several disorders, including malignancies, but its role is not well known. There are a group of inherited diseases affecting elastic fibres e.g. Marfan syndrome. It seems, that elastin degradation plays an important role in pathogenesis of some cardiovascular and pulmonary diseases. An enhanced elastolytic activity in malignancies is also observed. The data are rather sparse and above all concern lung and breast cancer. A trial estimating the role of elastin turnover in patients with hematologic malignancies, who underwent bone marrow transplantation, is now conducting in our department and preliminary results are promising.

Autoimmune Diseases↗

Genomic structure and organization of the high grade Myopia-2 locus (MYP2) critical region: mutation screening of 9 positional candidate genes.

PURPOSE: Myopia is a common complex eye disorder, with implications for blindness due to increased risk of retinal detachment, chorioretinal degeneration, premature cataracts, and glaucoma. A genomic interval of 2.2 centiMorgans (cM) was defined on chromosome band 18p11.31 using 7 families diagnosed with autosomal dominant high myopia and was designated the MYP2 locus. To characterize this region, we analyzed 9 known candidate genes localized to within the 2.2 cM interval by direct sequencing. METHODS: Using public databases, a physical map of the MYP2 interval was compiled. Gene expression studies in ocular tissues using complementary DNA library screens, microarray experiments, reverse transcription techniques, and expression data identified in external databases aided in prioritizing gene selection for screening. Coding regions, intron-exon boundaries and untranslated exons of all known genes [Clusterin-like 1 (CLUL1), elastin microfibril interfacer 2 (EMILIN2), lipin 2 (LPIN2), myomesin 1 (MYOM1), myosin regulatory light chain 3 (MRCL3), myosin regulatory light chain 2 (MRLC2), transforming growth beta-induced factor (TGIFbeta), large Drosophila homolog associated protein 1 (DLGAP1), and zinc finger protein 161 homolog (ZFP161)] were sequenced using genomic DNA samples from 9 affected and 6 unaffected MYP2 pedigree members, and from 5 external controls (4 unaffected and 1 affected). Gene sequence changes were compared to known variants from public single nucleotide polymorphism (SNP) databases. RESULTS: In total, 103 polymorphisms were found by direct sequencing; 10 were missense, 14 were silent, 26 were not translated, 49 were intronic, 1 insertion, and 3 were homozygous deletions. Twenty-seven polymorphisms were novel. Novel SNPs were submitted to the public database; observed frequencies were submitted for known SNPs. No sequence alterations segregated with the disease phenotype. CONCLUSIONS: Mutation analysis of 9 encoded positional candidate genes on MYP2 loci did not identify sequence alterations associated with the disease phenotype. Further studies of MYP2 candidate genes, including analysis of putative genes predicted in silico, are underway.

Chromosomes, Human, Pair 18↗

Three-dimensional modeling and molecular evolution of Matrix Cell Adhesion Molecule #2: MAT-CAM 2.

Our research group has recently reported the partial amino acid sequence of an aortic adventitial collagen-associated fibrillar protein, which may be the target of an autoimmune response in patients with abdommal aortic aneurysms (AAA). Its apparent molecular weight is approximately 40 kDa, so we have named it aneurysm-associated antigenic protein-40 kDa (AAAP-40). It has similarities to microfibril-associated glycoprotein-36 kDa (MAGP-36), which was reported by Kobayashi et al. to have a tissue distribution limited to the aorta in pig; unlike other microfibrillar proteins which appear to distribute ubiquitously throughout the body with elastin. Accordingly, if AAAP-40 is the human homolog of MAGP-36, it would explain how an autoimmune reaction against this protein might have consequences more or less limited to the aorta and its branches.

Journal Article↗

The spectrum of dermatological symptoms of pachydermoperiostosis (primary hypertrophic osteoarthropathy): a genetic, cytogenetic and ultrastructural study.

Pachydermoperiostosis (PDP) is characterized by finger clubbing, periostosis and peculiar skin involvement (pachydermia, seborrhea and folliculitis). The aim of our work was to determine the occurrence of dermatological symptoms in patients with PDP and their relatives, and to study ultrastructural skin changes in the complete and incomplete forms of the disease. A genetic and cytogenetic study was performed in order to identify the mechanism of transmission, to discover possible links to other genetic and non-genetic diseases and to determine the chromosomal complement and eventual chromosomal anomalies. Pachydermia was the most frequent skin alteration together with seborrhea; folliculitis was present in five patients. In the relatives mild pachydermia was detected in 2 out of 26, while seborrhea was present in 6 subjects. Light microscopic observation showed acanthotic epidermis and endothelial hyperplasia in the dermis with partial occlusion of the lumen, lymphohistiocytic infiltrate, and thickening and packing of collagen fibers. Electron microscopy showed fibroblast activation with increased fibrillogenic activity as shown by hypertrophic Golgi complexes and rough endoplasmatic reticulum with cisternae filled with microfibrils. Endothelial cells partially or completely occluded the capillary lumen and presented an increased amount of Weibel Palade bodies. These data show that skin involvement in PDP is a prominent feature, that sometimes these symptoms may also be present in their relatives, and that endothelial and fibroblast activation is present in the skin. Unfortunately the cytogenetic study did not provide any information about possible karyotype abnormalities.

Adult↗

Human vascular smooth muscle in culture. Growth and ultrastructure.

Mixed primary cultures of endothelial and smooth muscle cells were obtained from human umbilical cord vessels after prolonged collagenase digestion of their luminal surfaces. Morphologically homogeneous populations of vascular smooth muscle were then selectively isolated and subcultured for up to 16 weeks. Ultrastructurally, cultured cells were characterized by the presence of bundles of myofilaments with dense bodies similar to native umbilical vessel smooth muscle. Mature cultures developed a distinctive topographical organization consisting of bands of parallel cells and intertwined, multilayered areas. Elaborate intercellular attachments formed along contiguous cell surfaces. Large amounts of extracellular material accumulated, including amorphous substance, elastic fiber microfibrils, and 250- to 300-A,faintly-banded fibrils. In older cultures, focal proliferation, extracellular material secretion and cellular degeneration produced nodular protrusions somewhat resembling atherosclerotic lesions in vivo. Endothelium and smooth muscle cultured from this readily available human source may provide useful comparative material for pathophysiologic studies of vascular disease.

Culture Techniques↗

[Pseudoexfoliation syndrome--etiopatogenesis and clinical course].

Pseudoexfoliation syndrome is an ocular manifestation of a systemic elastosis. Exact etiology of this condition remains unknown. The basic pathogenetic concept of PEX is a pathological process of the extracellular matrix, characterized by the excessive production of an abnormal extracellular material which aggregates and accumulates and is not degraded in vivo. This material is produced primarily by the non pigmented epithelium of the ciliary body, the posterior iris pigment epithelium, and the preequatorial lens epithelium, while the corneal endothelium, trabecular cells, and vascular endothelia and smooth muscle cells of the iris have also been implicated. PEX material has a complex glycoprotein/proteoglycan, composition containing glycosaminoglycans (heparan sulfate, chondroitin sulfate, dermatan sulfate, hyaluronic acid). The prevailing presence of elastic fiber epitopes, mainly elastic microfibrillar components (elastin, vitronectin, amyloid P, fibrillin-1, LTBP-1), has led to the current theory explaining PEX as a type of elastosis, affecting especially elastic microfibrils. Ocular deposition of pseudoexfoliation material can lead to many complications in intraocular surgery like increased risk of zonular dehiscence, capsular rupture, vitreous loss during cataract extraction. Special attention is required before, during and after surgery.

Disease Progression↗

Properties of chitin synthetase in isolated chitosomes from yeast cells of Mucor rouxii.

Chitin synthetase was isolated and purified 120-fold from the supernatant fraction (54,500 X g) of broken yeast cells of Mucor rouxii. The purified preparations consisted mainly of chitin synthetase particles (chitosomes) with an average size larger than 7 X 10(6) daltons (by gel filtration) and an average sedimentation coefficient of 105 S. The samples also contained other enzyme complexes (fatty acid synthetase, pyruvate dehydrogenase, and, depending on method, ribosomes). Nearly all of the chitosomal chitin synthetase occurred in a zymogenic form that required proteolytic activation. In most properties, the chitosomal enzyme was similar to crude enzyme (54,000 X g sediment): kinetics, activation by proteases, response to metals, stimulation by N-acetylglucosamine, and inhibition by polyoxin or UDP. One mamor difference was the much greater stability of the chitosomal chitin synthetase zymogen against spontaneous activation and destruction. Product (chitin microfibril) and enzyme (chitin synthetase) remained associated in a complex that was readily separated by centrifugation.

Acetylglucosamine↗

Induction of cells with phenotypic features of neuronal cells by treatment with dibutyryl cyclic adenosine 3',5'-monophosphate in a human parotid gland adenocarcinoma cell line in culture.

A human parotid gland adenocarcinoma cell line, with an intercalated duct cell phenotype of the salivary gland and expression of vasoactive intestinal polypeptide and amylase, was cultivated in the presence of dibutyryl cyclic adenosine 3',5'-monophosphate (dB-cAMP). Morphological changes occurred; cells formed long cytoplasmic processes densely packed with ample microfibrils, as well as microtubules, and grew in a netlike appearance. In addition, it has been found by the immunofluorescence staining technique, immunoblotting, or immunoelectron microscopy that the cells treated with dB-cAMP express neurofilaments, neuron-specific enolase, synaptophysin, and HNK-1 antigen, as well as the alpha- and beta-chains of tubulin, whereas these antigens are not detected in untreated cells. The expression of vasoactive intestinal polypeptide detected diffusely in the cytoplasm of untreated cells was restricted to the cell membranes during the cultivation of cells in the presence of dB-cAMP, while expression of amylase persisted in the treated cells in a fashion similar to that in untreated cells. Moreover, both anchorage-independent and anchorage-dependent growth of the cells was markedly suppressed in the presence of dB-cAMP. After removal of dB-cAMP from the culture, the treated cells returned rapidly to the phenotype and growth rate of the untreated cells. These findings indicate that reversible conversion into cells with phenotypic features of neuronal cells of a human parotid adenocarcinoma cell line occurs in growth medium containing dB-cAMP.

Adenocarcinoma↗

Effect of epidermal growth factor on the cellular proliferation and phenotype of a neoplastic human salivary intercalated duct cell line or its derivatives.

A neoplastic human salivary intercalated duct cell line (HSG) and its derivatives, HSG with a myoepithelial cell phenotype (HSG-AZA1) and HSG with an acinar cell phenotype (HSG-AZA3), which were induced by 5-azacytidine treatment of HSG cells, were cultivated in the presence of epidermal growth factor (EGF). Morphological changes occurred; cells that were spindle shaped or stellate and had long cytoplasmic processes appeared in all of the treated cells. Major alterations, such as expression of neuron-specific enolase, neurofilaments, or synaptophysin as well as formation of neurite-like structures densely packed with microfibrils and microtubules, were observed in these cells with a phenotype similar to that of neuron-like cells. Both the anchorage-independent and anchorage-dependent growths of the treated HSG cells and only anchorage-dependent growth of the treated HSG-AZA1 cells were suppressed, whereas the anchorage-dependent growth of the treated HSG-AZA3 cells was enhanced according to the increasing concentrations of EGF. In addition, it has been found by statistical analysis that the growth of HSG cells in culture is suppressed and that of HSG-AZA3 cells in culture is stimulated according to the increasing concentrations of EGF added, whereas the growth of cultured HSG-AZA1 cells is hardly affected by EGF treatment. The HSG, HSG-AZA1, and HSG-AZA3 cell had 1.8 x 10(5), 1.83 x 10(5), and 1.34 x 10(5) EGF receptors, respectively. The amounts of the EGF internalized into cells were larger in HSG-AZA3 cells than in HSG or HSG-AZA1 cells. These findings indicate that the conversion of HSG, HSG-AZA1, and HSG-AZA3 cells into neuron-like cells occurs in growth medium containing EGF, with a concomitant modulation of the growth potentials of the cells which were examined in a different manner.

Amylases↗

Middermal elastolysis. Report of a case and immunohistochemical studies on the dermal distribution of fibrillin, vitronectin and amyloid P component.

A 39-year-old woman with demarcated wrinkled areas, histologically characterized by absence of elastic fibers in the middle and upper reticular dermis, is described. Immunoreactivity of vitronectin and amyloid P component, present at the periphery of elastic fibers in normal skin in adults, was absent from the middermis of lesional skin as were orcein stained fibers. C9 neoantigen immunoreactivity, associated with elastic fibers in sun-exposed skin of middle-aged and elderly individuals, was present in conjunction with elastic fibers in papillary and lower reticular dermis in lesional skin but was absent in the middermis. In contrast, a fibrillin immunoreactive network was present throughout the dermis, indicating that the elastin-associated microfibrils are retained in the absence of amorphous elastin in lesional skin of middermal elastolysis.

Adult↗

Patterns of actin filaments during cell shaping in developing mesophyll of wheat (Triticum aestivum L.).

Young leaves of wheat exhibit a smooth developmental gradient with meristematic cells at the base and highly differentiated cells at the tip. During differentiation, mesophyll cells attain a lobed outline resembling tube-shaped balloons with almost regularly spaced isthmi. Microfilament patterns in developing wheat mesophyll cells were investigated using fluorescent-labeled phalloidin. Various patterns were found, including delicate arrays of transversely oriented microfilaments in the cortex of the cytoplasm. A close correlation between changes in the patterns of cortical microfilaments, microtubules, cell wall microfibrils, and cell shape was observed. The fine arrays of transversely oriented microfilaments coaligned with bands of microtubules occurring during cell elongation. These bands were found beneath sites of intense wall deposition. It has recently been proposed that the resulting hoops of wall reinforcement prevent cell expansion in the corresponding regions and thus give rise to the peculiar cell shape. When cell expansion ceased, and the typical lobed cell shape was attained, a dense network of microfilaments was retained in the cytoplasm, which was in contrast to what has been described for the microtubular arrays.

Actin Cytoskeleton↗

Elastin in adenomatoid tumor. Light microscopic and electron microscopic study.

Elastin distribution was examined in adenomatoid tumors of the human genital tract. Two distinct patterns were identified: strongly positive or completely negative for elastin in the stroma, according to the case. Even in cases appearing to have very similar histologic features, the stroma was rich in elastin in some cases and was almost devoid of elastin in others. Electron microscopic examination with tannic acid staining revealed that the elastin in the stroma was composed mainly of amorphous material surrounded by a small amount of microfibrils and abundant collagen fibers. Fibroblasts were sparsely distributed in the stroma. Tumor cells displayed mesothelial cell-like features, such as abundant microvilli on the surface, numerous cytoplasmic organelles, several well-developed basal lamina. In some areas, elastin seemed to be formed by mesothelial cells. In some adenomatoid tumor cases, elastogenesis would be enhanced by activated mesothelial tumor cells as well as stromal fibroblasts.

Elastin↗

Cytopathic effects in primary epithelial cultures derived from the human prostate.

Normal, benign, and malignant human prostatic tissue was cultivated in vitro. Cytopathic effects in derived epithelial cells were examined. Light microscopy revealed polykaryocyte formation, vacuolation, cytoplasmic bridges and processes, nuclear inclusions, increased acid phosphatase activity at the cell membrane of polykaryocytes as compared to mononuclear cells, and cell rounding and clumping. Electron microscopy of the polykaryocytes showed nuclear membrane proliferation and protrusion, scarlike nuclear inclusions containing microfibrils, and virus-like particles similiar to viral nucleoids and nucleocapsids. The latter were also observed in the cytoplasm. The above alterations are similar to those induced by known herpesviruses. The significance of these changes, the possibility of the presence of a latent herpesvirus in the prostate, and its role in neoplastic disease are postulated.

Acid Phosphatase↗