Journal of submicroscopic cytology and pathology最新文献

The objective of the present study was to assess the possible toxic effects of chronic alcohol ingestion on the ultrastructure of the glandular epithelium of the prostate of the rodent Calomys callosus, in order to contribute to the understanding of the consequences of alcohol abuse for the morphology of the male reproductive apparatus. Sixteen adult animals aged three months were divided into two experimental groups. The control group received a solid diet and tap water, and the alcoholic group received the same solid diet and ethanol P.A. diluted 20% in water (v/v). After 120 days of treatment, all animals were anesthetized, weighed and sacrificed. At the end of treatment, mean body weight did not differ between control and alcoholic animals. The prostate epithelial cells of the alcoholic group showed intense atrophy and ultrastructural alterations such as the presence of lipid droplets, altered nuclei, ruptured mitochondrial cristae, and intense dilatation of the cisterns of the granular endoplasmic reticulum. It was concluded that 20% ethanol provokes marked lesions on the epithelium of the prostate probably interfering on the glandular secretion.

Nestin is a neuroepithelial precursor cell marker expressed in a variety of human cell types during development. However, no information exists on the expression of nestin in mature glomeruli as well as during the glomerular development. Here, we examined nestin expression in rat and human glomerular tissues in quiescent states using RT-PCR and immunohistochemical methods. Nestin mRNA was detected in the rat glomeruli in parallel with its expression in developing rat brains. In the normal mature rat glomeruli, WT-1 positive cells expressed nestin. Co-expression of nestin and vimentin was observed in mature rat podocytes. Immunoelectron microscopy revealed nestin localization in the cell bodies and primary processes of podocytes. A similar expression pattern was observed for vimentin. In matured glomeruli, nestin was not expressed by mesangial and endothelial cells. In the newborn rat, early developing glomeruli (metanephric cap, metanephric vesicle, comma-shaped vesicle and S-shaped body phases) expressed nestin. In the capillary loop stage, Bowman's capsules also expressed nestin. Immunoelectron microscopy demonstrated that developing podocytes and endothelial cells in S-shaped phase glomeruli expressed nestin. Additionally, in immature glomeruli, the mesangial cells in capillary stage of glomerulus also expressed nexin. As in the rat, WT-1 positive cells in human glomeruli also expressed nestin and immunoelectron microscopy confirmed nestin expression in human glomerular podocytes. These results reveal that in normal condition nestin is expressed in several glomerular cell types at early stage of development and becomes confined to podocytes in mature glomeruli, thus implicating nestin in podocyte functions.

Two types of autophagy in the podocytes were found in renal biopsy specimens by electron microscopy. Type I autophagy (about 1 microm in diameter) was found in 10 out of the 100 cases with renal diseases, and showed a condensed ribosome area with a limiting membrane. The origin of limiting membrane appeared to be from degenerated mitochondria. During type I autophagy formation, the thickness of limiting membrane changed from 5-6 nm to about 8-10 nm thickness. Type I autophagy did not transform to autophagosomes and autophagic vacuoles. On the other hand, many cases (90 out of the 100 cases) showed type II autophagy. Type II autophagy (3-8 microm in diameter) showed that many ribosomes were aggregated, formed condensed ribosome area, which always included many aggregated lipid droplets at first. Next, during the formation of autophagosome, rough ER connected to condensed ribosome area, and partly formed limiting membranes from dilated ER membrane. Finally, the limiting membrane of autophagic vacuoles was completely formed, and this membrane changed from about 5-6 nm to 8-10 nm thickness. Ribosomes and lipid droplets were resolved in autophagic vacuoles. Thus, type II autophagy might play a significant role in clearance of proteins and lipids in comparison with type I autophagy. The occurrence of type I autophagy in the renal biopsy specimens was not clearly associated with age, sex or pathological diagnosis. However, cases with type I autophagy may show a tendency to poor prognosis.

Biochemical studies demonstrate that the NO-releasing-aspirin derivative (NCX4016) stimulates soluble guanylate cyclase (sGC) activity and increases cyclic GMP (cGMP) in human platelet and monocytes by releasing NO. In the present study, an ultracytochemical technique for electron microscopy was used to investigate the effects of NCX4016 (2 mM) on sGC activity in rat thoracic aorta, using sodium nitroprusside (0.01 mM) as reference NO-donor. Guanylyl-imidodiphosphate sodium salt [Gpp(NH)p], a synthetic non-hydrolyzable analogue of GTP, was used as sGC substrate. NO-activated sGC released imidodiphosphate ions which were precipitated with lead ions, giving rise to deposits of electron-dense granules (reaction product). Ultracytochemistry allowed us to demonstrate that NCX4016 stimulated sGC activity in smooth muscle cells, and particularly in vascular endothelial cells, as sodium nitroprusside did. This result could explain the protective effects of chronic treatment with NCX4016 on aortic endothelium of diabetic rats demonstrated by scanning and transmission electron microscopy.

Malaria, a common health problem in certain parts of the world, has a considerable morbidity and mortality. This work reports under electron microscopy studies serious ultrastructural kidney damage such as extensive cytoplasmic vacuolation, vesiculation and autophagic vacuoles in proximal tubular cells. A thickened endothelial wall on peritubular capillary, interdigitation disorganization and significant decrease of their number in some areas were detected. Swollen rough endoplasmic reticulum, swollen mitochondria, and parasitized erythrocytes were observed. Many epithelial cells exhibited cytoplasmic areas of autophagia and a myelin-like form. A tubular cell presented severe cytoarchitecture alterations. Abundant lipid droplets were noticed. Almost total loss of interdigitations, rough endoplasmic reticulum vesiculation, peritubular capillaries with endothelial cells thickened cytoplasm, papillary processes projected to the lumen, and an inflammatory infiltrate of macrophages were also observed. These ultrastructural kidney changes could cause, on the basis of their clinical and pathologic expressions, a fat accumulation, an acute temporary reversible glomerulonephritis, a chronic progressive irreversible glomerulonephritis, and an acute renal failure (ARF).

Mitochondrial encephalomyopathies (MEs) are a group of clinically and genetically heterogeneous diseases. They can be caused by defects in both mitochondrial or nuclear coded genes. Their phenotypic expression is governed by unique biological phenomena such as the dual genetic control, mitotic segregation, heteroplasmy and threshold effects. Currently, the correct diagnosis of ME relies on a multidisciplinary approach which includes clinical information as well as laboratory data from muscle morphology, biochemistry and molecular genetics. Among the morphological methods, histology, histochemistry and electron microscopy were historically instrumental in the diagnosis of MEs. However, with the development of molecular genetics, the diagnostic value of morphology and of electron microscopy in particular have been questioned. The aim of the present review is to present a comparative assessment of the diagnostic contribution of histology, histochemistry and electron microscopy in a group of 48 patients with a diagnosis of ME.

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of 'piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

The light microscopy, histochemical and TEM studies of the epididymis and the vas deferens revealed the presence of PAS positive secretory granules in the epithelial cells lining the lumen of these organs. One dimensional SDS gel electrophoretic pattern of luminal fluid proteins and the total protein content of the testis, three regions of the epididymis and the vas deferens of the lizard, Mabuya carinata were studied during breeding and nonbreeding season of the reproductive cycle. During breeding season, 25 protein bands in the testicular luminal fluid, 26 in the anterior epididymal luminal fluid and 28 in the middle and posterior epididymal luminal fluid were found. Ten new protein bands appeared in the anterior epididymal region whereas five new protein bands appeared in the middle region of the epididymis indicating regional difference in protein secretions of the epididymis. Vas deferens luminal fluid showed the highest number of protein bands (32) and the highest total protein content (9.07 mg/ml) compared to the testis and the epididymis. Four new protein bands appeared in the vas deferens. Number of protein bands in the luminal fluids of testis, epididymis and the vas deferens were significantly reduced during nonbreeding season compared to those of the breeding season. Consistent with the decrease in the number of protein bands, there was a significant reduction in the total protein concentration in all the tissue samples during nonbreeding season. The results indicate seasonal differences in number of proteins secreted and quantity of proteins in the luminal fluid of male reproductive tract of M. carinata. This is the first study in reptiles revealing appearance of new proteins in epididymis, and vas deferens by conducting simultaneous electrophoretic profile of testicular, epididymal and vas deferens luminal contents.

Eggs from the bivalve mollusc Chamelea gallina were transfected in vitro. The p-GeneGrip gene construction that expresses the green fluorescent protein (GFP) was employed. It was necessary to remove the jelly coat which covers the egg surface for a successful transfection, and then 44.2% of gametes appeared transfected after using naked DNA. On the other hand, cationic liposomes (Lipofectamine) and neutral lipids (GenePORTER) were employed as gene vectors. After the employ of Lipofectamine 35.6% of eggs were transfected and 41.4% after using GenePORTER. Fluorescence analysis showed that the foreign gene appeared principally located in the egg cytoplasm, but laser confocal microscopy showed that it was also present in the nucleus. Furthermore, PCR analysis demonstrated that the foreign DNA appeared in the DNA extracted from the treated eggs. This simple method for the transfection of mollusc eggs would be interesting for future biotechnological applications in species of commercial interest.

The formation of feathers occurs by the transformation of the embryonic epidermis of feather filaments into keratinized barbules and barbs. The present ultrastructural study directly documents this transformation in chick and zebrafinch downfeathers and juvenile feathers. The transformation of the epidermis in the feather filament (downfeathers) or in the follicle (juvenile feathers) is similar. The change in cell shape of subperiderm or subsheath cells and surrounding barb vane ridge cells derives from the re-organization of the linear embryonic epithelium into barb ridges. In the latter the stratification of the outer and inner periderm, of the subperiderm/subsheath, and of the germinal layer of the embryonic epidermis is altered. While the external layers produce the sheath and barb vane ridge cells, subperiderm/subsheath cells are displaced into barbule plates that converge medially in the ramus area of the barb ridge. Cells in the barbule plates elongate into barbule and barb cortical cells by the synthesis of longitudinally oriented feather keratin bundles. In the mid-central area of the barb ridge (the ramus area) cells become polygonal and pile up. The external cells accumulate numerous keratin filaments forming cortical cells and are in contact with barbule cells. The above process also occurs in barb ridges of juvenile feathers and of adult feathers before molting. However, barb ridges produced within follicles of juveniles and adult feathers are longer than in downfeathers, and possess long rami. The incorporation of tritiated histidine in barbule and barb cortical cells has been studied by ultrastructural autoradiography. Most of the labeling is cytoplasmic or is associated with bundles of keratin but is not concentrated over keratin. This indicates that together with keratin possible histidine-rich keratin-associated proteins are produced during the elongation from subperiderm/subsheath to barbule/barb cells. Barb cortical cells merge with medullary cells of the ramus area. The latter accumulate lipids and few keratin bundles before degenerating into empty cells. Separation between barbule and barb cortical cells derives from the degeneration of barb vane ridge cells while separation between barb ridges derives from degeneration of cylindrical cells of marginal plates. These supportive cells incorporate less tritiated histidine than barbule/barb cells and their periderm granules are unlabelled with tritiated histidine. This indicates both that supportive cells are metabolically less active than feather-producing cells, and that putative histidine-rich proteins are only present in cells synthesizing feather keratin. Based on the morphogenesis of barb ridges a hypothesis on the evolution of downfeathers and pennaceous feathers is presented. From conical scales, thin hairy-like filaments were produced in which barb ridges were formed. The evolution of barb ridge morphogenesis with no fusion among barb ridges initially produced nak