A morphological and ultrastructural study of the paracloacal (Scent) glands of the marsupial Metachirus nudicaudatus Geoffroy, 1803

Morphological and ultrastructural features of the paracloacal glands of Metachirus nudicaudatus are described. Two pairs of glands, one on the right and the other on the left of the anal canal, are formed, each consisting of a major and a minor portion. Their wall is made up of three layers: a mucosal, a muscular and an external capsule. The inner one is a mucosa the epithelium of which contains holocrine cells characterized by lipid droplets and intermediate filaments. The surrounding vascular lamina propria contains flattened tubular apocrine glands whose epithelial cells contain abundant endoplasmic reticulum, prominent Golgi complexes and numerous secretory granules. The middle layer is formed by skeletal striated muscle and the outer (third layer) consists of dense connective tissue. Each gland originates from a single duct. Transverse sections show that each duct, except in the female major gland, is in fact formed by a duct system. One of these ducts comes from the central cavity, lined by holocrine epithelium, and the others result from the branched tubular glands of the lamina propria.

Peripheral giant cell granuloma. An immunohistochemical and ultrastructural study.

OBJECTIVE: To study the nature of multinucleated and mononuclear cells from peripheral giant cell granuloma (PGCG). MATERIALS AND METHODS: Formalin-fixed, paraffin-embedded sections of 40 cases of PGCG were immunohistochemically stained for vimentin, alpha I-antichymotrypsin, CD68, S-100 protein, lysozyme, leucocyte common antigen (LCA), factor VIII-related antigen and muscle cell actin. Six cases of PGCG were also studied by transmission electron microscopy. RESULTS: Vimentin, alpha I-antichymotrypsin and CD68 were expressed in both the mononuclear and multinucleated giant cells. Dendritic mononuclear cells, positive for S-100 protein, were noted in 67.5% of the lesions, whereas lysozyme and leucocyte common antigen were detected in occasional mononuclear cells. Ultrastructural examination showed mononuclear cells with signs of phagocytosis and sometimes interdigitations with similar cells. Others presented non-specific characteristics and the third type exhibited cytoplasmic processes and occasional Birbeck granules. Some multinucleated giant cells showed oval nuclei, abundant mitochondria and granular endoplasmic reticulum whereas others presented with irregular nuclei and a great number of cytoplasmic vacuoles. CONCLUSIONS: Immunohistochemical and ultrastructural results suggest that PGCGs of the jaws are composed mainly of cells of the mononuclear phagocyte system and that Langerhans cells are present in two thirds of the lesions.

Gap junctions between mast cells and fibroblasts in the developing avian eye

Mast cells are present in the eye of chick embryos from the 14th day onward, displaying metachromatic granules, mainly in the iris anterior surface and pectinate ligament. Ultrastructurally these cells show electron-dense granules and a few thin and short cytoplasmic projections in close contact with fibroblasts. Sometimes these contacts are extensive, with long fibroblast projections partially involving the mast cells. Gap junctions between mast cells and fibroblasts are observed only in the eyes of 16- and 20-day-old embryos. These intercellular specializations are represented by a close apposition of cytoplasmic membranes with an extension up to 300 nm. Gap junctions between mast cells and fibroblasts were not observed previously in vivo or in vitro, although in vitro studies have shown that a number of functionally critical interactions may occur between these cells. Our morphological findings suggest that, in vivo, fibroblasts interact with mast cells and may influence their maturation.

Ultrastructural study of the coagulating gland of Wistar rats submitted to experimental chronic alcohol ingestion

Morphological changes of coagulating gland of Wistar rats submitted to the experimental chronic alcoholism were noted. Ultrastructurally, it was observed reduction of the granular endoplasmic reticulum cisternae and nuclei with basal position and irregular shape. The epithelial cells presented pycnotic nuclei and the mitochondrial cristae disappeared, characteristic of cellular degeneration.

Features of myoneural junctions in the extraocular muscles of the opossum (Didelphis albiventris)

The myoneural junctions present in the opossum (Didelphis albiventris) extraocular muscles were studied through histochemical and ultrastructural techniques. Three different types of junction were shown: two single types, 'en grappe' and 'en plaque', present in the middle third of the muscle fibers; and one multiple type, more rare and observed in the distal third of the muscle.

Laminin-binding epitope on gp43 from Paracoccidioides brasiliensis is recognized by a monoclonal antibody raised against Staphylococcus aureus laminin receptor

Adhesion is regarded as an important step in the pathogenesis of several microorganisms. Thus, the ability to recognize extracellular matrix proteins, such as laminin or fibronectin, has been correlated with invasiveness. Studying the already characterized laminin-binding protein of Paracoccidioides brasiliensis, the 43 kDa glycoprotein (gp43), we evaluated whether MAb 1.H12, raised against the laminin-binding protein from Staphylococcus aureus, cross-reacts with that fungal protein. By immunoblot analysis we show that MAb 1.H12 recognizes gp43. This interaction is able to inhibit the laminin-mediated adhesion to epithelial cells as well as the P. brasiliensis infection in vivo. Moreover, through immunoenzymatic assays, we show that MAb 1.H12 recognizes gp43 in solid phase and that this interaction is partially inhibited by the addition of anti-gp43 MAbs. These results show that MAb 1.H12 recognizes the gp43, suggesting the presence of an epitope similar to those found in the other laminin-binding proteins from phylogenetically very distant cells. These findings reinforce the possibility of evolutionary conservation of such epitopes.

Ultrastructural and morphometric aspects of ageing in the retinal capillaries of rats

Age-related morphological, ultrastructural and morphometric changes in the capillaries of the superficial and deep plexuses of the rat retina were studied in animals aged from 3 to 15 months. Our results suggest that age-related morphological alterations start occurring in the retina of rats at about 12 months of age. Increased glycogen deposits, pinocytotic vesicles, residual bodies and cell debris were observed in both the endothelial and pericytic cells of 12- and 15-month-old animals. In addition, heterogeneous osmiophilic accumulations, electron-transparent spaces were observed in the basement membrane as well as projections of the basement membrane towards the neighboring cells. Morphometric examination of the two vascular plexuses studied did not show differences in the area of the endothelial or pericytic cells, basement membrane or vascular lumen between rats of different ages.

Ultrastructural study of the lateral lobe of the prostate of Wistar rats submitted to experimental chronic alcohol ingestion

Alcoholism is considered today one of the most serious social and medical problems. Different investigators have demonstrated that chronic exposure to alcohol causes morphological and physiological changes in the testes and in the accessory sex glands, in the prostate in particular. In the present study, experimental rats were divided into groups receiving sugar cane brandy diluted to 30° G.L. (30%, v/v) and Purina ration ad libitum for 60, 120, 180, 240 and 300 days, respectively. At the end of each period the animals were sacrificed and the lateral lobe of the prostate was collected for histological examination. The results showed atrophied epithelium, accumulation of lipidic droplets and rupture of the microvilli that line the cell surface facing the lumen of cells of the secretory epithelium of the lateral lobe of the prostate.

Comparative structural analysis of neuromuscular junctions in mice at different ages

The authors studied the histochemical and ultrastructural modifications that occur in the neuromuscular junctions (NMJ) of fibularis longus muscles of mice with an age range of 3 to 21 months. Twenty-four male and female animals were killed at 3, 5, 14 and 21 months of age: 7 of them at 3 months, 4 of them at 5 month, 9 at 14 months and 4 at 21 months. The fibularis longus muscles were processed and their NMJ examined with the transmission electron microscope. The most relevant changes were associated with the degeneration and retraction of terminal axons, i.e., axons poor in synaptic vesicles with degenerated mitochondria, and exhibiting multivesicular bodies and vacuoles; exposed and widened junctional folds and cytoplasmic processes of Schwann cells located in the synaptic gutter. The presence of lysosomes or lipofuchsin in the juxtajunctional sarcoplasm was also noted. These observations suggest that the phenomena of retraction and budding occur in the NMJ with advancing age, with a predominance of events associated with degeneration, leading to profound changes in NMJ shape.

Ultrastructural changes on the epithelial cells of uterine tubes of Wistar rats after chronic ethanol ingestion

The present paper describes the morphological alterations of the epithelial layer of the uterine tubes of rats submitted to experimental chronic alcoholism using anatomical, histological, ultrastructural and morphometric methods. Sixty adult rats (Rattus norvegicus albinus) at the same age (3 months) and with a mean body weight of 228 g were divided into two groups. The control group received solid diet (Purina rat chow) and tap water ad libitum. The alcoholic group received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30° Gay Lussac (v/v). After periods of 90, 180 and 270 days of treatment animals at normal estrus were anaesthetised with ethyl ether, weighed and sacrificed. Subsequently, the uterine tubes were dissected, weighed and prepared for TEM and SEM methods. The final mean body weights were similar in the control and alcoholic groups. The morphometric analysis showed no difference between control and alcoholic epithelial height. The alcoholic animals showed ultrastructural alterations: intense lipid droplet and lysosomes accumulation, dilated rough endoplasmic reticulum cisternae and vacuolization in both periods of treatment. It was concluded that alcohol acts as a toxin on the epithelial layer of the uterine tubes of rats.

Three-dimensional reconstruction of fracture surfaces: Area matching algorithms for automatic parallax measurements

This paper describes two solutions for systematic measurement of surface elevation that can be used for both profile and surface reconstructions for quantitative fractography case studies. The first one is developed under Khoros graphical interface environment. It consists of an adaption of the almost classical area matching algorithm, that is based on cross-correlation operations, to the well-known method of parallax measurements from stereo pairs. A normalization function was created to avoid false cross-correlation peaks, driving to the true window best matching solution at each region analyzed on both stereo projections. Some limitations to the use of scanning electron microscopy and the types of surface patterns are also discussed. The second algorithm is based on a spatial correlation function. This solution is implemented under the NIH Image macro programming, combining a good representation for low contrast regions and many improvements on overall user interface and performance. Its advantages and limitations are also presented.

Tracheobronchial consequences of the use of heat and moisture exchangers in dogs

Purpose: To determine the effect of heat and moisture exchange (HME) on the tracheobronchial tree (TBT) using a unidirectional anesthesic circuit with or without CO2 absorber and high or low fresh gas flow (FGF), in dogs. Methods: Thirty-two dogs were randomly allocated to four groups: G1 (n = 8) valvular circuit without CO2 absorber and high FGF (5 L·min-1); G2 (n = 8) as G1 with HME; G3 (n = 8) circuit with CO2 absorber with a low FGF (1 L·min-1); G4 (n = 8) as G3 with HME. Anesthesia was induced and maintained with pentobarbital. Tympanic temperature (TT), inhaled gas temperature (IGT), relative (RH) and absolute humidity (AH) of inhaled gas were measured at 15 (control), 60, 120 and 180 min of controlled ventilation. Dogs were euthanized and biopsies in the areas of TBT were performed by scanning electron microscopy. Results: The G2 and G4 groups showed the highest AH (>20 mgH2O·L-1) and G1 the lowest (< 10 mgH2O·L-1) and G3 was intermediate (<20 mgH2O·L-1) (P < 0.01). There was no difference of TT and IGT among groups. Alterations of the mucociliary system were greatest in G1, least in G2 and G4, and intermediate in G3. Conclusion: In dogs, introduction of HME to a unidirectional anesthetic circuit with/without CO2 absorber and high or low FGF preserved humidity of inspired gases. HME attenuated but did not prevent alterations of the mucociliary system of the TBT.

Cell death in ovarioles causes permanent sterility in Frieseomelitta varia worker bees

Frieseomelitta varia worker bees do not lay eggs even when living in queenless colonies, a condition that favors ovary development and oviposition in the majority of highly social bees. The permanent sterility of these worker bees was initially attributed to a failure in ovary morphogenesis and differentiation. Using transmission electron microscopy we found that at the beginning of the pupal phase the ovaries of F. varia workers are formed by four ovarioles, each of them composed of 1) a terminal filament at the apex of the ovarioles, containing juxtaposed and irregularly shaped cells, 2) a germarium with clusters of cystocytes and prefollicular cells showing long cytoplasmic projections that envelop the cystocyte clusters, 3) fusiform interfollicular and basal stalk precursor cells, and 4) globular, irregularly contoured basal cells with large nuclei. However, during the pupal phase an accentuated and progressive process of cell death takes place in the ovarioles. The dying cells are characterized by large membrane bodies, electron-dense apoptotic bodies, vacuoles, vesiculation, secondary lysosomes, enlarged rough endoplasmic reticulum cisternae, swollen mitochondria, pycnotic nuclei, masses of chromatin adjacent to the convoluted nuclear envelope, and nucleoli showing signs of fragmentation. Cell death continues in ovarioles even after the emergence of the workers. Once they become nurse bees, the ovaries have become transformed into a cell mass in which structurally organized ovarioles can no longer be identified. In F. varia workers, ovariole cell death most certainly is part of the program of caste differentiation.

Apoptosis in the early developing periodontium of rat molars

Development of the periodontium involves a series of complex steps that result in the formation of root dentine, cementum, bone and fibres of the ligament. These precisely controlled and timed events require the participation of the enamel organ derived epithelial cells of Hertwig's (HRS) and ectomesenchymal cells of the dental follicle. These events involve rapid turnover of the tissues and cells, including disappearance of epithelial cells of HRS. Thus, it seemed likely to us that programmed cell death (apoptosis) may play a role in the development of the periodontium. Fragments of first molars, obtained from 14- and 29-day-old rats, were fixed in glutaraldehyde-formaldehyde and processed for light and electron microscopy. For the TUNEL method for detection of apoptosis, specimens were fixed in 4% formaldehyde and embedded in paraffin. Results confirmed that epithelial cells of HRS maintain a close relationship with the forming dentine root, and that they may become trapped in the dentino-cemental junction. Some of the epithelial cells exhibited ultrastructural features which are consistent with the interpretation that they were undergoing programmed cell death, i.e. apoptosis. Periodontal fibroblast-like cells showed typical images of apoptosis and engulfed apoptotic bodies. TUNEL positive structures were present in all corresponding regions. It seems therefore that apoptosis of epithelial cells of HRS and fibroblast-like cells of the periodontal ligament constitutes an integral part of the developmental process of the tissues of the periodontium. (C) 2000 Wiley-Liss, Inc.

Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia

Toward the end of the larval phase (pre-pupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries. (C) 2000 Wiley-Liss, Inc.