Morphometric study of the postnatal growth of the parotid gland of the mouse

The growth of the mouse parotid glands during 7 and 35 days of postnatal life was studied by morphometric methods. The mass of the gland, the volume of each morphological compartment, and the cell number in each compartment were evaluated. The data obtained for each evaluated dimension were adjusted by an exponential equation, of the type Y = a.e KX, thus permitting the calculation of their mean duplication time (T D), i.e., an estimation of their growth rate. Analysis of the results showed a marked 1,424% increase in the gland mass during the whole studied period, with T D = 7.10 days. This growth occurred by increases in absolute volume of acini, intercalated ducts, striated ducts, excretory ducts and stroma, with percentual increases of 3,048%, 417%, 2,662%, 2,594% and 367%, respectively, and T Ds of 5.62, 11.71, 5.55, 5.47 and 14.45 days, respectively. Analysis of the cell number growth in each compartment showed increases of 1,904%, 285%, 1,228%, 1,090% and 286%, respectively, and T Ds of 6.62, 20.40, 7.19, 7.26 and 14.51 days, respectively. Based on the present results, we concluded that the growth of the mouse parotid glands from day 7 to day 35 of age occurred by intense cell accumulation, mainly in the acini, striated ducts and excretory ducts, with a growth rate sensibly higher than that of the intercalated ducts and stroma.

Cytologic analysis of alterations induced by smoking and by alcohol consumption

Objective: To analyze cytologically the buccal mucosa of smoking and nonsmoking volunteers to determine what cellular changes are induced by cigarettes and alcohol consumption. Study Design: In order to evaluate cellular changes induced by smoking and alcohol consumption, exfoliative cytology was used for the analysis of mucosal smears obtained from the buccal mucosa of 25 smokers and 25 nonsmokers. The number of cigarettes consumed, duration of smoking, presence or absence of alcohol ingestion, ingested alcohol dose and frequency of consumption, and most frequently used type of alcoholic beverage were determined using a questionnaire. Three smears from each individual stained by the Papanicolaou method were analyzed quantitatively and qualitatively under a light microscope by 2 experienced examiners in terms of inflammatory and dysplastic alterations and of the degree of epithelial maturation. Results: Although numerous alterations were observed in smokers they corresponded up to only Papanicolaou class II and were not significantly different from nonsmokers (Mann-Whitney and χ 2 tests, p < 0.05). A higher proportion of inflammatory cells (polymorphonuclear and mononuclear cells) were obtained from smokers as compared to nonsmokers, while the proportion of bacteria was similar in the 2 groups. Conclusion: The findings indicate that even after a short period of cigarette use and alcohol consumption, inflammatory alterations were detectable on exfoliative cytology of the buccal mucosa in a young group, demonstrating the usefulness of cytology for early detection in smokers. © The International Academy of Cytology.

Morphological alterations on the prostate of Calomys callosus submitted to chronic ethanol ingestion

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.

Ultrastructural features of the lining epithelium of the vas deferens of Agouti paca

The epididymal epithelium of Agouti paca, a wild South American rodent, was basically formed by principal and basal cells. Principal cells were closely related to processes of adsorptive endocytosis, phasefluid endocytosis and also secretion originating from their cytoplasmic ultrastructural features. Principal cells were also characterized by the presence of vesicles of several shapes, sizes and internalized content occurring in smaller pits, pale small vesicles next to the apical brush border of microvillus, as well as coated vesicles, smooth surface vesicles and great vesicles. Multivesicular bodies, endosomes and lysosomes were mainly observed in supranuclear position. Moreover, presence of an apocrine secretory process was demonstrated by the occurrence of apical cytoplasmic expansions projecting into the vas deferens luminal compartment. Basal flattened cells without luminal surface contact occurred next to the basement membrane of the ductus, and did no exhibit special ultrastructural features.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

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.

Exposição de culturas de fibroblastos de pterígios primários e recidivados à mitomicina c e ao 5-fluorouracil

Purpose: To evaluate the proliferation of the fibroblasts from primary and recurrent pterygium in culture of cells, after the exposure to mitomycin C and to 5-fluorouracil. Methods: Cultures of fibroblasts from primary and recurrent pterygiuns had been carried through. The cells of the third passage were exposed to mitomycin C 0.4% during 3 minutes and to 5-fluorouracil 25mg/ml that was kept in the nutritional medium. After 3, 6, 12 and 18 days of the exposure, cell countings were made in hemocytometer, in triplicate, with a control not treated with the drugs for each day. The data were submitted to statistical analysis.Results: Mitomicyn C had homogeneous behavior in the primary and recurrent groups, which inhibited the cellular proliferation since the first counting day. However, with 5-fluorouracil, the proliferation inhibition was verified only after the third day of evaluation (in the second counting day) in the recurrent group, and since the first counting day in the primary group. At the end of the experimental period, 5-fluorouracil and mitomycin C were equally efficient in the inhibition of the fibroblasts from primary and recurrent pterygium proliferation. In the controls not exposed, the cell's numbers were increasing during the experimental time.Conclusion: Mitomycin and 5-fluorouracil are equally able to inhibit fibroblasts proliferation from primary and recurrent pterygium Tenon's capsule in cell culture.

Study of chromosomal and nucleolar aspects in testes of Nysius californicus (Heteroptera: Lygaeidae)

In Nysius californicus (family Lygaeidae, subfamily Orsillinae), a pest commonly known as the seed bug, the chromosome complement is 2n = 16 (12A + 2m + XY), testes are formed by seven seminiferous tubules covered by an orange-colored membrane, and spermatogenesis is cystic. At prophase, sex chromosomes are heteropycnotic and autosomes usually show a chiasma. At metaphase, sex chromosomes along with microchromosomes may be seen located at the center of a ring formed by the remaining autosomes. A characteristic specific of N. californicus was the presence of nucleolar material observed from the cystic cell to the completely differentiated spermatozoon. Variations in size, shape and location of the nucleolar material occur during this process, denoting a variable degree of activity in the different stages. ©FUNPEC-RP.

Bone healing in critical-size defects treated with bioactive glass/calcium sulfate: A histologic and histometric study in rat calvaria

Objective: The purpose of this study was to analyze histologically the influence of bioactive glass (BG) with or without a calcium sulfate (CS) barrier on bone healing in surgically created critical-size defects (CSD) in rat calvaria. Material and methods: A CSD was made in each calvarium of 48 rats. They were divided into three groups: C (control): blood clot only; BG: defect filled with BG; and BG/CS: defect filled with BG covered by a CS barrier. Animals were euthanized at 4 or 12 weeks. Formation of new bone was evaluated histomorphometrically. Results: No defect completely regenerated with bone. BG particles were observed in Groups BG and BG/CS at both periods of analysis. The thickness throughout the healing area in Groups BG and BG/CS was similar to the original calvarium, while Group C presented a thin connective tissue in the center of the defect in both periods of analysis. At 4 weeks, Groups C and BG/CS presented significantly more bone formation than Group BG. No significant differences were found between Groups C and BG/CS. At 12 weeks, no significant differences in the amount of bone formation were observed among the three groups. When comparing 4 and 12 weeks, there was a significant increase in new bone formation within groups BG and BG/CS, but not C. Conclusion: BG particles, used with or without a CS barrier, maintained the volume and contour of the area grafted in CSD. However, they did not lead to a significant difference in bone formation when compared with control at 12 weeks post-operative. © 2007 Blackwell Munksgaard.

Sporotrichosis in an HIV-positive man with oral lesions: A case report

Background: Sporotrichosis is a granulomatous fungal infection caused by Sporothrix schenckii, which frequently causes cutaneous or lymphocutaneous lesions and rarely has oral manifestations. Case: A 38-year-old, white, HIV-positive man complained of a 5.0-cm, symptomatic, ulcerated lesion with thin, superficial granulation in the soft palate extending to the uvula. Exfoliative cytology of this oral lesion showed chronic granulomatous inflammatory alterations and extracellular fungal structures consisting of periodic acid-Schiff-positive budding cells and spherical or elongated (cigar bodies) free spore forms. Conclusion: The clinical and cytologic findings allowed the diagnosis of sporotrichosis, demonstrating the importance of cytodiagnosis in fungal diseases. © The International Academy of Cytology.

Effects of basic fibroblast growth factor on density and morphology of fibroblasts grown on root surfaces with or without conditioning with tetracycline or EDTA.

A study was conducted to evaluate in vitro the effect of root surface conditioning with basic fibroblast growth factor (b-FGF) on morphology and proliferation of fibroblasts. Three experimental groups were used: non-treated, and treated with 50 microg or 125 microg b-FGF/ml. The dentin samples in each group were divided into subgroups according to the chemical treatment received before application of b-FGF: none, or conditioned with tetracycline-HCl or EDTA. After contact with b-FGF for 5 min, the samples were incubated for 24 h with 1 ml of culture medium containing 1 x 10(5) cells/ml plus 1 ml of culture medium alone. The samples were then subjected to routine preparation for SEM, and random fields were photographed. Three calibrated and blind examiners performed the assessment of morphology and density according to two index systems. Classification and regression trees indicated that the root surfaces treated with 125 microg b-FGF and previously conditioned with tetracycline-HCl or EDTA presented a morphology more suggestive of cellular adhesion and viability (P = 0.004). The density of fibroblasts on samples previously conditioned with EDTA, regardless of treatment with b-FGF, was significantly higher than in the other groups (P < 0.001). The present findings suggest that topical application of b-FGF has a positive influence on both the density and morphology of fibroblasts.

Susceptibility/resistance of Anticarsia gemmatalis larvae to its nucleopolyhedrovirus (AgMNPV): Structural study of the peritrophic membrane

This investigation compares the peritrophic membrane (PM) morphology along the midgut of susceptible (SL) and resistant (RL) Anticarsia gemmatalis larvae to the AgMNPV. The PM increased the thickness from the anterior to the posterior midgut region in both insects strain; however, the intensity of FITC-WGA reaction of the PM in the RL were greater than in SL. The PM in RL was ultrastructurally constituted by several layers of fibrous/vesicular materials in comparison with the few ones in SL. Our results showed that the structure of PM in the RL could be one of the resistance barriers to AgMNPV. © 2007.

Avoiding leukocyte contamination and early platelet activation in platelet-rich plasma.

The objective of this study was to describe a new platelet-rich plasma (PRP) protocol with a reduced concentration of leukocytes and intact platelets. We collected 8 mL of venous blood (VB) from marginal ear veins of 10 male New Zealand white rabbits in acid dextrose citrate Vacutainer tubes. Tubes were centrifuged at 302g for 10 minutes. All plasma was collected in plastic tubes to avoid buffy-coat contamination and centrifuged at 2862g for 5 minutes. A 10% calcium chloride activator (10 PRP:2 CaCl2) was added to the lower third of this plasma (PRP), and the PRP gel was obtained. Mean platelet count was 317.7 x 10(3) +/- 39.9/microL in VB and 1344.9 x 10(3) +/- 347.5/microL in PRP. Leukocyte counts were 3.96 x 10(3) +/- 2.01/microL and 0.46 x 10(3) +/- 0.45/microL in VB and PRP, respectively. Mean platelet enrichment was 327.4 +/- 97.8%. All differences were statistically significant (P > .05). This protocol is practical and reproducible, resulting in a high concentration of intact platelets to help tissue repair and low levels of leukocytes.

Ophthalmic patterns of captive brown brocket deer (Mazama gouazoubira)

Captive brown brocket deer (Mazama gouazoubira) were manually restrained to assess tear production by the Schirmer tear test I to measure intraocular pressure by applanation tonometry, to examine ocular conjunctival epithelial cells via cytologic and histologic samples, and to survey ocular conjunctival microflora by microbiologic culture. The mean value for the Schirmer tear test I was 8.9 ± 1.8 mm/min, and the mean intraocular pressure was 15.3 ± 3.1 mm Hg. Conjunctival epithelium contained stratified pavimentous layers of cells, and the microflora consisted of predominantly gram-positive bacteria. Copyright 2007 by American Association of Zoo Veterinarians.

Aspirado traqueal de cavalos clinicamente sadios da raça quarto de milha após prova de três tambores

The aim of the present study was to evaluate through endoscopy the tracheal aspiration cytology in twenty seven Quarter Horses from Curitiba and surroundings, following the Three Barrel Competittion. Upper respiratory tract secretion was obtained by tracheal aspiration using a polyethylene catheter introduced through the endoscopic fiberoptic working channel, at the level of tracheal bifurcation. Cytologic slides were prepared by smear and stained by diff-quick technique and the differential was performed in 500 cells counting by 1,000X optic microscopy. None of the horses presented abnormality, including epixtasis, at the clinical examination. However, hemosiderophages were detected at cytology in three animals, suggesting that some may be suffering of subclinical pulmonary hemorrhage. Differential cell counting of tracheal aspiration results were, in average: 44.09 ± 35.68% of epithelial cells; 1.10 ± 2.18% of Globet cells; 23.10 ± 35.93% of neutrophils; 0.13 ± 0.37% of lymphocytes; 0.91 ± 2.81% of eosinophils; 30.57 ± 23.62% of macrophages and 0.13 + 0.93% of hemosiderophages. In conclusion, based in the present study, the evaluation of cellular populations with the tracheal aspiration may offer important additional information to the clinician, particularly about the inflammatory processes of lower respiratory tract and pulmonary bleeding.

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Background: Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils. Methods: Eosinophils were purified using a percoll gradient followed byimmunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry. Results: At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils. Conclusion: Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion. © 2008 Lintomen et al; licensee BioMed Central Ltd.