Diazepam administration prevents testosterone decrease and lipofuscin accumulation in testis of mouse exposed to chronic noise stress.

Lipofuscin is an autofluorescent and undegradable material, which accumulates in tissues during ageing and under different types of stress. Among these, oxidative stress represents a major trigger for lipofuscin formation. However, prolonged noise exposure is also an effective stressful stimuli. Diazepam may inhibit lipofuscinogenesis in liver and prevent the noise-induced reduction of the steroidogenesis in the adrenal gland. The aim of the study was to ascertain whether chronic noise exposure causes lipofuscin accumulation in mouse testis, and to evaluate the effects of diazepam administration. Eight-week old mice were either exposed for 6 weeks (6 h day(-1)) to white-noise (group A), or received diazepam (3 mg kg(-1), i.p.) before noise exposures (group B), while a further group was used as control (group C). Light fluorescence and transmission electron microscopy revealed lipofuscin in large amounts in the Leydig cells in mice of group A, which concomitantly had low serum testosterone levels; pre-treatment with diazepam occluded both effects. The present study indicates that: (i) chronic noise exposure causes lipofuscin accumulation at the level of the Leydig cells and a decrease in testosterone; (ii) all these effects are suppressed by pre-treatment with diazepam. As the Leydig cells represent the only cellular type of the interstitial testicular tissue having peripheral benzodiazepine receptors, these results could be explained by the capacity of the peripheral benzodiazepine receptors to prevent reactive oxygen species damage and to increase the resistance of these cells to oxidative stress.

Inflammatory mediators and eosinophilia in atopic and non-atopic patients with nasal polyposis.

Nasal polyps are characterized by eosinophilic infiltration and presence of inflammatory mediators, such as total IgE, eosinophil cationic protein (ECP) and cytokines. The role of atopy in nasal polyp pathogenesis is still unclear. Therefore, we evaluated serum IgE levels, nasal mucus concentrations of ECP and cytokines and the number of infiltrating eosinophils in nasal tissue of polyps from atopic and non-atopic patients. Samples were obtained from a randomized population of 31 patients with nasal polyposis having endonasal sinus surgery and of 13 control subjects undergone corrective surgery of the nasal septum. On the basis of medical history of allergy, positive skin-prick tests and total IgE levels, patients with polyposis were divided in atopic (n = 13) and non-atopic (n = 18) patients. We determined levels of IgE in blood, ECP and cytokines (IL-4, IL-6, IL-8, IFN-gamma and IL-2) in nasal mucus, and number of infiltrating eosinophils in nasal tissue. The concentrations of total IgE, ECP, IL-4 and IL-8 and eosinophilia were significantly higher in all patients with nasal polyps compared with controls. Inside, all patients with nasal polyposis showed lower levels of IL-6, IFN-gamma and IL-2 compared with controls. The atopic patients showed significant differences when compared with non-atopic patients for the higher concentrations of total IgE (698.80+/-322.24 vs. 279.63+/-234.11; P < 0.0001) and IL-8 (1437.2 pg/ml+/-1250.7 vs. 605.5 pg/ml+/-481.1; P < 0.015). These findings suggest that inflammation still remains the major factor in the etiology of nasal polyposis and show different levels of inflammatory mediators into atopic and non-atopic patients.

Ankyloglossia: a morphofunctional investigation in children.

OBJECTIVES: To provide diagnostic criteria for ankyloglossia in children by anatomical measurements; to investigate the correlation between severity of ankyloglossia and a series of morphofunctional findings; to evaluate the potential mismatch between a clinical suspect of ankyloglossia and the authentic anatomical diagnosis. DESIGN: Two different techniques of anatomical measurements and a clinical evaluation of a series of morphofunctional findings were performed. SUBJECTS AND METHODS: In 200 children referred for evaluation and treatment of tongue-tie, the length of the frenulum and the interincisal distance were measured in maximum opening of the mouth and with the tip of the tongue touching the palatal papilla. Occlusion, type of bite, tongue resting position, swallowing mechanism, oral floor mobility, frenulum insertion modality and speech were investigated. Any correlation between these morphofunctional findings and anatomical measures was investigated. RESULTS: Children with a frenulum length more than 2 cm and an interincisal distance of more than 2.3 cm were normal. In both measurements, significant correlations among mean values and other variables were observed. Moreover, three levels -- mild, moderate and severe -- of ankyloglossia were assessed. CONCLUSIONS: Length of frenulum and interincisal distance allow an assessment of severity of ankyloglossia in children. Ankyloglossia was not associated with infantile swallowing.

Distribution of 3-nitrotyrosine in the nasal polyps of atopic patients.

OBJECTIVE: To investigate whether formation of nitrotyrosine in the nasal polyps of atopic patients occurs. STUDY DESIGN: A nonrandomized, retrospective, controlled qualitative and quantitative study. METHODS: Nasal polyp tissue samples were acquired from 12 atopic patients. Control fragments of nasal mucosa were taken from 10 patients undergoing corrective surgery of the nasal septum. For routine histologic examinations, hematoxylin-eosin staining was used. Low-magnification microscopy was designed to yield pathologic characteristics and high magnification to quantify the number of eosinophils in the subepithelial connective tissue. Presence of nitrotyrosine was assessed by immunohistochemical method. RESULTS: Hematoxylin-eosin staining revealed presence of numerous eosinophils in the epithelium and in the subepithelial connective tissue. All polyps were characterized by epithelial damage. Nitrotyrosine was present in the eosinophils, in the ciliated cell, and in cells of the damaged epithelium. Goblet cells, glands, and vessels were found to be negative. No significant differences concerning the localization of nitrotyrosine were recognized among the examined nasal polyps. CONCLUSIONS: Nitrotyrosine immunohistochemical staining in nasal-polyp tissues suggested the existence of progressive epithelium injury caused by peroxynitrite. Consequences of peroxynitrite formation in eosinophils remain to be precisely established. The lack of nitrotyrosine in glands and blood vessels indicated that peroxynitrite does not have a significant role in the vascular and glandular dysfunction of nasal polyps.

Ultrastructural localization of the NADPH-diaphorase activity in the Leydig cells of aging mice.

Recently, it has been shown that nitric oxide may inhibit the Leydig cell steroidogenesis. The present paper describes, by means of NADPH-diaphorase histochemistry, the ultrastructural localization of the enzyme nitric oxide synthase in the Leydig cells of young adult and aging mice. In the young adult mice, the enzymatic reaction was mainly located in the mitochondria and in some clustered cisternae of the smooth endoplasmic reticulum. The nuclear envelope was faintly labeled. In the aging mice, most Leydig cells showed an enhanced enzymatic reaction. Labeled mitochondria were increased in number, and labeled areas of the smooth endoplasmic reticulum were more numerous and extended. In addition, a strong enzymatic reaction was recognized in the nuclear envelope. We conjecture that the impaired steroidogenesis observed in the testis of aging mammals might, at least in part, depend on the increased nitric oxide production in the Leydig cells.

Ultracytochemical localization of the NADPH-d activity in the human nasal respiratory mucosa in vasomotor rhinitis.

OBJECTIVES: Description of the ultrastructural localization of nitric oxide synthase in the blood vessels of the nasal respiratory mucosa in patients with vasomotor rhinitis. STUDY DESIGN: This research was conducted on seven patients--men and women, ages 20 to 45 years--suffering from vasomotor rhinitis and who had undergone surgical therapy for reduction of the inferior turbinates. METHODS: To study the ultrastructural localization of nitric oxide synthase, NADPH-diaphorase cytochemistry was employed. Samples of the nasal mucosa were obtained from inferior turbinates. RESULTS: The endothelial cells of the arterioles, capillaries, venules and cavernous sinuses revealed a distribution of the enzymatic activity similar to that found in unaffected subjects. A strong enzymatic activity was recognized in the smooth muscle cells of the cavernous sinuses. The smooth muscle cells of arterioles and venules were generally found to be negative to enzymatic reaction. CONCLUSIONS: This study suggests that the vascular disorders of the vasomotor rhinitis depend, at least in part, from nitric oxide synthase induction in the smooth muscle cells of the cavernous sinuses.

Ultrastructural localization of NADPH-diaphorase activity in the endothelial cells of human nasal respiratory mucosa.

The cavernous sinuses are the most peculiar feature of the nasal angioarchitecture, due to their ability to retain a large quantity of blood in reply to a variety of topical and systemic stimuli. Recently, nitric oxide (NO) has seemed to be crucially involved in the nasal vascular regulation. The distribution of NO-synthase (NOS), the enzyme that catalyzes the formation of NO, was studied in the endothelium of nasal blood vessels by the ultracytochemical detection of reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) enzymic activity. The endothelium of the cavernous sinuses appeared strongly positive, whereas the endothelium of arterioles was occasionally labeled. The endothelial cells of capillaries and venules were found to be NADPH-d negative. The strong enzymic activity observed in the cavernous sinuses suggests a major role of NO in the capacitance vessels compared to the resistance vessels. The hypothesis of a reciprocal inhibition between the NOS enzymic pathways present in the respiratory epithelium and in the endothelium of cavernous sinuses is put forward. The nasal disorders characterized by anomalous vasomotility and vascular permeability could be caused in part by the irregular control of these complex interactions.

Cytochemical localisation of the NADPH diaphorase activity in the Leydig cells of the mouse.

The distribution of the NADPH diaphorase activity was studied in mouse Leydig cells by means of light and electron microscopy. When observed by the light microscope, most Leydig cells appeared intensely stained; a few cells (about 10%) showed a slightly positive or apparently negative reaction. The inhibitory effects of N(G)-nitro-L-arginine and iodonium diphenyl on frozen sections suggest the colocalisation of NADPH diaphorase reaction with nitric oxide synthase. The ultrastructural study revealed that all the Leydig cells were positively stained for NADPH diaphorase; however, a small number of cells displayed weak enzymatic activity. The reaction product was located in the mitochondria, smooth endoplasmic reticulum and lipidic vacuoles, and the nuclear envelope was also stained. The possible meaning of the NADPH diaphorase activity in the Leydig cells of mice was discussed.

Ultrastructure of the supporting cells in the paratympanic organ of chicken, Gallus gallus domesticus.

The paratympanic organ is a specialized sensory organ of birds located in the medial wall of the tympanic cavity. It possesses a sensory epithelium formed by type II hair cells and supporting cells. The supporting cells are tall, narrow units that extend from the basement membrane to the free epithelial surface. They show a fine structure characterized by numerous mitochondria, a conspicuous Golgi complex and a well-developed RER. Moreover, some uncommon structures, probably formed by heaped RER cisternae, are frequently present in the cytoplasm. Adjacent supporting cells are connected by numerous and extensive gap junctions; moreover, small gap junctions between hair cell and supporting cells are to be found. The possible mechanical and metabolical functions of the paratympanic organ supporting cells are discussed.

The ultrastructural localization of NADPH-diaphorase enzymatic activity in the Leydig cells of mouse. effects of ethanol administration.

The effects of the chronic ethanol treatment on the NOS-related NADPH-diaphorase activity were described in the mouse Leydig cells by means of transmission electron microscope. The recovery of the Leydig cells was also examined during a period of four weeks. About 10% of the Leydig cells showed various degrees of morphological alterations, consisting in increased number of lipid droplets, rarefaction and vacuolization of the cytoplasmic matrix. Other groups of Leydig cells (about 10%) revealed evident signs of degeneration. The NADPH-d activity was reduced both in apparently normal and injured Leydig cells and a moderate enzymatic reaction was only detected in the smooth endoplasmic reticulum. A week after the treatment an increased number of the degenerating Leydig cells and a further reduction of the enzymatic reaction were observed. Then, the Leydig cells showed a progressive recovery and four weeks after the treatment they exhibited a normal morphology and NADPH-d enzymatic reaction. These results demonstrated for the first time the inhibition of NOS activity in the Leydig cells after chronic ethanol administration.

Ultrastructural demonstration of the NADPH-diaphorase activity in peritubular myoid cells and fibroblasts of the lamina propria of the mouse seminiferous tubules.

The NOS-related NADPH-diaphorase activity was studied by transmission electron microscopy in the peritubular myoid cells and fibroblasts of normal mouse testis. The reaction product was observed on the membranes of the endoplasmic reticulum, on the Golgi apparatus and nuclear envelope. The peritubular myoid cells and fibroblasts showed similar ultracytochemical features; the intensity of the enzymatic reaction was suggestive of an important role of the NOS/cGMP enzymatic system in these cells. Some hypotheses on the role of NO in the peritubular myoid cells and fibroblasts are proposed.

Epixenosomes: peculiar epibionts of the ciliate Euplotidium itoi: morphological and functional cell compartmentalization.

As already reported, epixenosomes have a complex cell organization although they lack membranous organelles and a nuclear envelope. Different morphological compartments have been evidenced: (1) a space (IM) between the 2 external membranes in which acid phosphatase has been observed; (2) an apical, electrondense zone (DZ) not delimited by a membrane or thin envelope, containing DNA and basic proteins; (3) a roundish body (RB), 200 nm in diameter, containing reserve polysaccharides and enzymes, in particular, peroxidase; (4) an extrusive apparatus (EA) consisting of a ribbon with smooth surfaces in which lectin binding sites are present, and immersed in a complex matrix different from the remaining cytoplasm. Along its internal and external limits, active adenylate cyclase has been evidenced. At its top, a 'ring' of peroxidase is present; (5) a basket surrounding the extrusive apparatus probably consisting of tubulin microtubules. The cytochemical data, especially the precise localization of the enzymes shown here, indicate that in epixenosomes, a functional compartmentalization corresponds to a structural complexity, in spite of the absence of internal true membranes.