Journal of submicroscopic cytology and pathology最新文献

To examine the calcification of implanted hydrogel IOL by X-ray microanalysis, we compared conventional transmission electron microscopy (TEM) with low-vacuum scanning electron microscopy (SEM). We also compared metal coating with non metal coating in low-vacuum SEM. Calcification of IOL showed deposits which were located in the superficial substance of lens. In conventional TEM and X-ray microanalysis, calcium, phosphate and silicon were detected in the deposits. In low-vacuum SEM, the deposits detected in metal coating were calcium, phosphorus, sodium and magnesium, but not silicon. However, in non metal coating, the deposits contained not only calcium, phosphorus, silicon, sodium and magnesium, but also fluoride, aluminum and argentums. It was concluded that in conventional TEM where a specimen is fixed and dehydrated in ethanol, various elements leak out. On the other hand, when a specimen is coated with carbon and gold palladium for SEM, light elements might not be detected in X-ray microanalysis. Low-vacuum SEM preparation does not need metal coating and low-vacuum SEM appears to provide a highly efficient method for X-ray microanalysis.

A retrospective study to detect specific Y chromosome microdeletions and to evaluate sperm ultrastructural characteristics in infertile men was set up. We selected 219 infertile men referred to Regional Referral Center for Male Infertility, Siena, Italy for semen analysis from January 1999 to April 2004. Family history, lymphocyte karyotype determination, Y microdeletion screening, physical examination, hormonal assays, semen analysis were carried out. Sperm concentration and progressive motility, ultrastructural analysis of sperm organelles, PCR amplification of sequence tagged sites for Y microdeletion screening were performed. Different Y-chromosome deletions were found, mainly in the AZFb and AZFc regions. Severe alterations of sperm ultrastructure, affecting whole sperm population, were detected in carriers of Y-deletions. Our data confirms the highest frequency of Y deletions in azoospermic patients. In all other patients with Y microdeletions, sperm ultrastructural defects affected the whole sperm population and were mainly related to apoptosis or immaturity.

Different steps of sperm activation such as acrosomal reaction and capacitation are described in details. The molecules involved in sperm-egg interaction are also reported.

mUBPy is a deubiquitinating enzyme expressed preferentially in male germ cells and neurons. Recently, mUBPy has been shown to be involved in the down-regulation of growth factor receptors. In mouse spermatozoa mUBPy interacts with the sperm-specific molecular chaperone MSJ-1 and associates with the proteasome. The ubiquitin/proteasome system plays a key role during spermatogenesis to yield functional spermatozoa. Immunoelectron microscopy has been here used to localize both mUBPy and MSJ-1 in mouse spermatozoa. mUBPy and MSJ-1 label the cytoplasmic side of the acrosomal membrane and the centrosome, two sperm structures fundamental for a successful fertilization. In vitro protein interaction assay reveals that mUBPy is able to bind gamma-tubulin, a centrosomal protein marker. This protein interaction has been confirmed in vivo by double protein immunolabelling in spermatogenic cells. Upon the grounds of these findings and in the light of recent acquisition on the centrosome biology, we suggest that mUBPy could have a key role during mouse fertilization and propose mUBPy as a novel centrosomal component.

Cortical biopsies of 18 patients with clinical diagnosis of congenital hydrocephalus, brain trauma, and vascular anomaly were examined with the transmission electron microscope to study the distinct and overlapped morphological cell types of nerve cell death in the human edematous cerebral cortex. The nerve cells showed lobulated and shrunken nucleus, irregular enlargement and fragmentation of perinuclear cistern, with areas of apparently intact nuclear pore complexes alternating with regions of nuclear pore complex disassembly. The nucleolus appears unaltered in moderate edema and with distorted nucleolar subcompartments in severe edema. Most nonpyramidal nerve cells, astrocytes and oligodendrocytes underwent an oncotic-apoptotic-necrotic continuum featured by swollen nucleoplasm, cytoplasm, and cell organelles, chromatin condensation and marginalization, and formation of apoptotic bodies. In a lesser proportion other nonpyramidal nerve cells, astrocytes and oligodendrocytes only showed apoptosis or oncosis. Autophagic cell death characterized by presence of autophagic vacuoles of lysosomal origin was rarely seen. The above findings suggest that different mechanisms of nerve cell death occur in the human edematous cerebral cortex related with brain trauma, congenital hydrocephalus, vascular anomaly, and their anoxic-ischemic conditions. An oncotic-apoptotic continuum process leading to necrosis predominates in human cerebral cortex nerve cell populations. The nerve cell death is discussed in relation with the severity of brain edema, anoxic-ischemic conditions of brain parenchyma, oxidative stress, glutamate excitotoxicity, calcium overload, and caspase dependent and independent mechanisms.

The sperm 'round head' defect, also known as globozoospermia, is an uncommon alteration of sperm morphology generally characterised by 100% round headed sperm totally lacking an acrosome. This alteration is a genetic sperm defect as demonstrated by analysing the incidence of these alterations in a population of infertile men showing a history of consanguinity and cases belonging to the same family. Ultrastructural characteristics and meiotic segregation in spermatozoa from two patients affected by 'round head' sperm defect were investigated. The sperm quality was examined by light and transmission electron microscopy (TEM) and fluorescence in situ hybridization (FISH) analysis was performed in order to investigate the meiotic behavior of chromosomes namely gonosomes and chromosome 18. TEM analysis, mathematically elaborated, clearly diagnosed the 'round head' genetic sperm defect and highlighted at the same time the presence of other phenotypic alterations belonging to pathologies such as immaturity, apoptosis and necrosis. It is possible to hypothesize that round headed sperm could be a 'weak phenotype' allowing the sperm pathologies to overlap with a sperm defect of genetic origin, further compromising fertilizing potential. FISH analysis revealed a positive correlation between globozoospermia and higher disomies of sex chromosomes and diploidies suggesting a higher risk of creating an aneuploid embryo after intracytoplasmic sperm injection.

The present ultrastructural study describes the formation of feather ramification in developing juvenile feathers of the zebrafinch, a small passeraceous bird. The study stresses the importance of the detailed knowledge on the cell structure of barb ridges for the understanding of feather development and evolution. Feather formation depends on the morphogenesis of long barb ridges, in which cells are displaced into lateral barbule plates and a medial barb cells region. These cells merge into long chains and form a syncitium organized in a ramified structure that preserves the original cell disposition within the barb ridge. Barb vane ridge cells surround barb and barbule cells. Barbules separate after the degeneration of barb vane ridge cells. In barbule cells the formation of hooklets resembles the process of formation of climbing setae of digital pads of some lizards. The cytoplasm of barb vane ridge cells is localized among tile-like overlapped barbule cells that form barbule chains, and maintains a serrated outline. When barb vane ridge cells degenerate among keratinized barbules, keratinized hooklets remain. Hooklets allow the ordered grasping of barbules to form a close and planar vane of feathers. The rachis of juvenile feathers seems to be formed from the fusion of two or more barb ridges localized in the dorsal part of the follicle, but the process of fusion is unclear. Juvenile and adult feathers contain the same type of feather keratin present in downfeathers: this indicates that stem cells for the regeneration of a new feather remain in the follicle after shedding of downfeathers. The presence of embryonic organelles (periderm granules) in barb vane ridge cells of juvenile feathers further indicates that also stem cells for the regeneration of the latter cells remain in the follicle. Molting feathers are therefore derived from stem cells. The permanence of stem cells in the follicle and the modulation of barb ridges dimension and fusion into different patterns allow the production of different feather morphotypes such as contour, filoplumes, semiplumes, and bristles.

The morphology of spermatozoon of Sceliphron fistularium is very similar to that described for bees. In particular, the response to E-PTA stains is similar to that observed in corbiculated Apidae, especially Meliponini bees. Spermatozoa measure 285 microm and are composed of 1) a bilayered acrosome (acrosomal vesicle and perforatorium); 2) a homogeneous and compact nucleus; 3) a 9+9+2 axoneme; 4) a rod-shaped centriolar adjunct; 5) two asymmetrical mitochondrial derivatives with paracrystalline material exclusively in the larger one, and 6) two accessory bodies. Only the accessory microtubules of axoneme and the paracrystalline material are E-PTA positive. Comparison of S. fistularium sperm to data on Hymenoptera corroborates their proximity with bees.

Trypanosoma musculi, a protozoan parasite specific to mouse, was cultured in vitro in the presence of spleen-derived adherent cells. T. musculi co-cultured with adherent cells survived and proliferated indefinitely as long as cellular contact was retained. Scanning and transmission electron microscopy confirmed intimate membrane-to-membrane contact between the adherent cells and parasites. Cellular contact, therefore, seemed to be essential for trypanosomal survival and growth. Immunocytochemical studies demonstrated intense fibroblast growth factor (FGF) activity in adherent cells, and FGFR-2 in associated trypanosomes. BioPorter Lucifer yellow protein delivery reagent studies demonstrated that Lucifer yellow transfected into fibroblast was incorporated into associated trypanosomes. The results suggest the existence of viable channels reminiscent of gap junctions between associated cells. Such transfer of low molecular weight molecules might represent antiapoptotic metabolic factors that support survival of adherent trypanosomes in vitro. Immunocytochemical studies also detected connexin-32 and connexin-43 in the cytoplasm of fibroblasts and associated trypanosomes, however, restriction of connexons to trypanosome/fibroblast adherent sites was not observed. Western blots confirmed the presence of connexin protein molecules in trypanosomes.

This paper describes the ultrastructure of the commoner myofibroblastic tumours and tumour-like lesions. The objective is to complement mainstream pathology texts, which have concentrated on the clinical and light microscopy features of these lesions and which have arguably but understandably somewhat neglected electron microscopy as an ancillary diagnostic tool and a technique for investigating tumour cell biology. Ultrastructural features are described of nodular fasciitis, the myofibromatoses (including Dupuytren's disease), inflammatory myofibroblastic tumour, post-operative spindle cell nodule, fibroma of tendon sheath, fibrous pseudotumour, benign fibrous histiocytoma, atypical fibroxanthoma, dermatofibrosarcoma protuberans, myofibrosarcoma (myofibroblastic sarcoma), malignant fibrous histiocytoma (pleomorphic myofibrosarcoma), epithelioid sarcoma and spindle-cell carcinoma. Fibrosarcoma and leiomyosarcoma are illustrated for comparison. The fibronexus is emphasised as an important marker for the most confident diagnosis of myofibrosarcoma. Some pathologists accept a light microscope definition, which includes alpha-smooth-muscle actin positivity, h-caldesmon negativity and, in some cases, desmin positivity. Caution in the interpretation of desmin staining in a possible myofibroblastic lesion is urged, since, in combination with an ultrastructurally identified lamina, it more probably suggests true smooth-muscle differentiation. Myofibroblastoma and angiomyofibroblastoma are examples of tumours argued on the basis of ultrastructural findings (sometimes in combination with desmin staining) to be primitively differentiated smooth-muscle cell rather than myofibroblastic proliferations.