Action of colchicine on hepatic schistosomal granuloma

Amorphous material and altered collagen fragments within dilated secretory vesicles and cisternae of fibroblast cytoplasm were the main ultrastructural changes seen in hepatic periovular granulomas formed in mice infected with Schistosoma mansoni and treated with colchicine. Despite promoting ultrastructural changes in the fibroblasts found in hepatic periovular granulomas, colchicine administration to infected mice did not significantly change the light microscopic appearance of the hepatic schistosomal lesions, did not diminish the amount of total hepatic collagen, and did not change the collagen isotypes in the granulomas, as observed after a comparative study with non-colchicine treated infected control mice. When administered to mice two weeks after curative treatment of schistosomiasis with praziquantel, colchicine did not seem to increase extracellular collagen degradation or to induce a more rapid resorption of hepatic periovular granulomas, although still promoting ultrastructura alterations in fibroblasts.

New methods for the diagnosis of Babesia bigemina infection

Accurate diagnosis of Babesia bigemina infection, an economically important tick-transmitted protozoan parasite of cattle, is essential in the management of disease control and in epidemiological studies. The currentlyused methods of diagnosis are blood smear examination and serological tests which include agglutination and immunofluorescence tests. These testes have been used the fild but because they lack sensitivity and specificity, never and improved methods of diagnosis are being developed. The quantitative buffy coat (OBC) method, using microhaematocrit tubes and acridine orange staining allows rapid and quicker diagnosis of B. bigemina and other blood parasites compared to light microscopic examination of stained smears. Parasite specific monoclonal antibodies have been used in antigen/antibody capture enzymelinked immunosorbent assays with grater sensitivity and specificity than previously described serological tests. Similary, DNA probes, derived from a repetitive sequence of the B. bigemina genome, offer a method of detecting very small numbers of parasites which are undetectable by conventional microscopy. An extrachromosomal DNA element, present in all the tick-borne protozoan parasites so far tested, provides an accurate means of diferentiating mixed parasite populations in infected animals. These improved methods will greatly facilitate epidemiological studies.

Intracortical connectivity of layer VI pyramidal cells in the barrel cortex of mice

ABSTRACT : The whisker-to-barrel pathway of rodents is formed by a series of somatotopic projections from the mystacial whisker follicles to the layer IV of the primary somatosensory cortex such that each follicle corresponds to a cluster of cortical neurons called barrel. Barrels are present in layer IV but form part of functional columns that comprise the entire depth of the somatosensory cortex. Interestingly, the cortex of the barrelless mouse strain (BRL) is organized such a manner that thalamocortical afferents do not remodel their projections in layer IV and barrels fail to appear. Nevertheless, functionally, a columnar organization persists, indicating that functional columns are not only provided by thalamocortical projections and layer IV cells. Since in the visual cortex of cats, layer VI cells contribute to the response properties of layer IV neurons, we wonder whether layer VI pyramidal cells could contribute to the columnar organization of the primary somatosensory cortex of mice. To address -this question, we morphologically analyzed the distribution of intracortical axon collaterals of layer VI neurons after in-vivo juxtacellular injections of biocytin in the C2 barrel column. Injected hemispheres were tangentially serial cut and intracortical collaterals of individual layer VI neurons were reconstructed at the light microscopic level. The position of axonal boutons was recorded to evaluate the distribution of presumed synaptic contacts. In normal (NOR) mice, cluster analysis shows that layer VI pyramidal cells can be classified in four statistically different clusters of neurons. Moreover, we assume that two classes are formed by cortico-cortical neurons and two classes are formed by cortico-thalamic neurons. Looking at the direction of the main axon in the white matter, we noticed that its orientation correlates perfectly with the type of neuron: cortico-cortical neurons send main axon medially whereas cortico-thalamic neurons send main axon laterally. Performing the same study in the BRL strain, we showed that the BRL mutation affects layer VI pyramidal cells tangentially and radially: the effects of the mutation are illustrated by a significant decrease of the index of colurnnarization and a significant decrease of percentage of boutons in granular and supragranular layers comparing to NOR neurons. In spite of these differences, the same four classes of layer VI neurons have been found in BRL mice. Using a tangential analysis of the boutons distribution, we showed that putative synapses are distributed mainly in the C2 barrel column. This was observed for each layer, type of neuron, cluster or strain, indicating that layer VI pyramidal cells could participate to the functional columnar organization of the barrel cortex. To determine post-synaptic partners of layer VI neurons in layer IV, we conducted an ultrastructural analysis of layer VI-to-IV contacts. We showed that synapses principally occur on spines and spiny dendritic shafts, supposed to belong to excitatory neurons. We furthermore showed that pre-synaptic elements are significantly different between en passant and terminaux contacts, which support hypothesis that terminaux boutons should show longer duration of facilitation than en passant boutons. RÉSUMÉ : Le «whisker-to-barrel pathway» des rongeurs est caractérisé par une série de projections somatotopiques depuis les follicules des moustaches ('whiskers') jusqu'à la couche IV de l'aire somatosensorielle primaire, de telle façon que chaque follicule corresponde à un groupe de neurones corticaux appelés tonneaux (`barrels'). Les tonneaux sont seulement présents en couche IV mais font partie de colonnes fonctionnelles qui s'étendent sur toute la profondeur du cortex somatosensoriel. Chez les souris mutantes barrelless (BRL), le cortex somatosensoriel est organisé de façon telle que lés afférences thalamocorticales ne remodellent pas leurs projections en couche IV et que les tonneaux n'apparaissent pas. Fonctionnellement, pourtant, une organisation en colonnes persiste, ce qui indique que les colonnes fonctionnelles ne sont pas uniquement produites par les projections thalamocorticales et par les cellules de la couche IV. Puisque les cellules de la couche VI contribuent à influencer les réponses des cellules de la couche IV dans le cortex visuel du chat, nous nous sommes demandé si ces cellules ne pourraient pas aussi contribuer à l'organisation en colonnes du cortex somatosensoriel primaire de la souris. Pour répondre à cette question, nous avons analysé de façon morphologique la distribution intracorticale des collatéraux axonaux de neurones de la couche VI. Suite à des injections juxtacellulaires de biocytine in-vivo dans la colonne C2, les hémisphères cérébraux ont été tangentiellement coupés en série et les collatéraux intracorticaux des neurones de la couche VI ont été reconstruits en microscopie optique. La position des boutons axonaux a aussi été enregistrée pour évaluer la distribution des contacts synpptiques potentiels. Chez les souris NOR, une analyse multivariée montre que les cellules pyramidales de la couche VI sont distribuées en quatre classes. Deux de ces classes sont probablement formées de neurons cortico-corticaux, alors que les deux autres sont probablement formées de neurones corticothalamiques. En observant la direction de l'axone principal dans la matière blanche, nous avons noté que son orientation est parfaitement corrélée avec le type supposé de neurone : les neurones corticocorticaux envoient leurs axones principaux médiallement, alors que les neurons cortico-thalamiques envoient leurs axones principaux latéralement. En menant la même étude chez les souris BRL, nous avons montré que la mutation affecte les cellules pyramidales de la couche VI de façon tangentielle, mais aussi radiaire : les effets de 1a mutation se traduisent par une diminution significative de l'index de « columnarization » et de la connectivité en couches granulaire et supragranulaire. Malgré ces différences, les quatre mêmes classes de neurones ont été retrouvées. En utilisant une analyse tangentielle de la distribution des boutons, nous avons montré que les synapses potentielles sont distribuées principalement dans la colonne C2. Cette observation a été faite dans chaque couche, chaque type de neurones, chaque classe de neurones et chaque souche de souris, indicant que les cellules de la couche VI participent certainement à l'organisation en colonne du cortex somatosensoriel. Pour déterminer les partenaires post-synaptiques des cellules de la couche VI en couche IV, nous avons conduit une analyse ultrastructurelle de ces contacts. Nous avons montré que les synapses interviennent principalement sur les épines et sur les dendrites supposés appartenir à des cellules excitatrices. Nous avons aussi montré que les éléments pré-synaptiques de ces synapses sont significativement differents selon le type de bouton, en passant ou terminal, ce qui supporte l'hypothèse que les boutons terminaux seraient capables d'une plus longue facilitation.

Protective effect of simvastatin in the cyclophosphamide-induced hemohrragic cystitis in rats

Cyclophosphamide (CYP) is an antineoplastic agent used for the treatment of many neoplastic and inflammatory diseases. Hemorrhagic cystitis is a frequent side effect of CYP. Several studies show that simvastatin has important pleiotropic (anti-inflammatory and immunomodulatory) effects. The purpose of the study was to investigate the effect of simvastatin on bladder, ureter and kidney injury caused by CYP. Methods: Adult male Wistar rats were randomly divided into three groups. The CYP/SIM group received simvastatin microemulsion by gavage during 7 days (10 mg/kg body wt) before the administration of CYP and the CYP/SAL group rats received saline 0.9%. The control rats were not treated. After that, all rats were treated with a single dose of CYP 200 mg/kg body wt intraperitoneally. The rats were killed 24 h after CYP administration. Plasma cytokines (TNF-a, IL-1b, IL-6) were measured by ELISA. Macro and light microscopic study was performed in the bladder, kidney and ureter. Results: In the bladders of CYP/SIMV treated rats edema of lamina propria with epithelial and sub-epithelial hemorrhage were lower than in CYP/SAL treated rats. The scores for macroscopic and microscopic evaluation of bladder and ureter were significantly lower in CYP/SIMV rats than in CYP/SAL rats. The kidney was not affected. The expression of TNF-a, IL-1b and IL-6 was significatly lower in CF/SINV rats (164.8±22, 44.8±8 and 52.4±13) than in CF/SAL rats (378.5±66, 122.9±26 e 123.6±18), respectively. Conclusion: The results of the current study suggest that simvastatin pretreatment attenuated CYP-induced urotelium inflammation and decreased the activities of cytokines

A microestrutura e o desenvolvimento dos órgãos genitais masculinos de preás (Galea spixii) criados em cativeiro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Human pulpal response to direct pulp capping with an adhesive system

Purpose: To evaluate clinically and microscopically the human pulp response when directly capped with an adhesive system or calcium hydroxide over short (9-12 days) and long (53-204 days) experimental periods. Materials and Methods: Fifty-one sound human premolars scheduled for orthodontic extraction, had their pulp horns gently exposed with a diamond point. Debris in the pulp wound was washed out with a sterile saline solution. The pulps were then capped with either an adhesive system (Scotchbond Multi-Purpose Plus) or calcium hydroxide. All teeth were subsequently restored with resin-based composite (Z-100) according to the manufacturer's instructions. After the experimental periods, the teeth were extracted and processed for light microscopic examination. Results: Short-term: the pulp tissue capped with SBMP-P exhibited dilated and congested blood vessels associated with a mo;derate inflammatory response and blanching of pulp cell nuclei. Long-term: no evidence of healing and bridge formation was observed. A persistent mild inflammatory pulp response was present. Micro-abscesses were detected in three cases associated with bacterial infiltration. Calcium hydroxide stimulated early pulp repair and dentin bridging which extended into the longest period.

Morphologic alterations resulting from denervation of the diaphragm in rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morphometric study of the midgut epithelium in larvae of Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae)

A larva de Diatraea saccharalis Fabricius (broca-da-cana) tem grande interesse econômico, pois afeta o cultivo e aproveitamento industrial da cana-de-açúcar. Entretanto, poucos são os estudos sobre a morfologia interna desse inseto. O objetivo deste trabalho foi estudar, morfometricamente, o seu epitélio intestinal, ao longo de seu comprimento, visando caracterizar regiões estruturalmente diferentes. O intestino médio de larvas no último instar foi subdividido em três regiões: proximal, mediana e distal e os fragmentos foram processados para observação em microscopia de luz. Os cortes histológicos foram analisados em sistema computadorizado de análise de imagens para medir comprimento, largura e área do epitélio, das diferentes células epiteliais, dos seus respectivos núcleos e do lúmen intestinal. Os dados obtidos foram submetidos ao teste estatístico de Kruskal-Wallis e à análise multivariada. Nossos resultados mostraram que o intestino médio apresentou-se constituído, morfometricamente, por duas diferentes regiões, proximal e distal; a região mediana apresentou valores coincidentes tanto com a região proximal quanto com a distal, sugerindo ser região intermediária. As células epiteliais (colunares, caliciformes e regenerativas), quando avaliadas pela análise estatística multivariada, não apresentaram diferença morfométrica nas diferentes regiões do intestino médio. Entretanto, a análise de variância, realizada para variáveis isoladas, mostrou que as células regenerativas apresentaram maior variabilidade morfométrica.

Effect of high laryngeal mask airway intracuff pressure on the laryngopharyngeal mucosa of dogs

Objectives: To evaluate the effect of increased of laryngeal mask airway (LMA) intracuff pressures on the laryngopharyngeal mucosa. Study Design: Animal model. Methods: Sixteen mixed-breed dogs were randomly allocated to two groups, G1 (intracuff volume, 30 mL; n = 8) and G2 (intracuff volume, 54 mt; n = 8), to produce, respectively, high or very high intracuff pressures. Anesthesia was induced and maintained with pentobarbital. Intracuff pressures were measured immediately after insertion and inflation of a No. 4 laryngeal mask airway (LMA) and 30, 60, 90, and 120 minutes thereafter. The dogs were euthanized, and biopsy specimens from eight predetermined areas of the laryngopharynx in contact with LMA cuff were collected for light microscopic (LM) and scanning electron microscopic (SEM) examination. Results: Initial LMA cuff inflation in G1 and G2 resulted in intracuff pressures of 119 mm Hg +/- 4 mm Hg and 235 mm Hg +/- 13 mm Hg, respectively. Over a 2-hour period, the intracuff pressure decreased significantly in G1 (P < .001) and G2 (P < .01), and there was a significant difference between the groups over time (P < .001). The LM study of laryngopharyngeal mucosa in both groups showed mild congestion in the subepithelial layer. There were no differences between the groups (P > .10) or among the areas sampled (P > .10). In some areas of G2, the SEM study showed epithelial desquamation that was more intense than that in GI. Conclusions: the increase in LMA intracuff pressure caused only mild alterations in the laryngopharyngeal mucosa of the dog.

The effects of low laser irradiation on angiogenesis in injured rat tibiae

The influence of He-Ne laser radiation on the formation of new blood vessels in the bone marrow compartment of a regenerating area of the mid-cortical diaphysis of the tibiae of young adult rats was studied. A small hole was surgically made with a dentistry burr in the tibia and the injured area received a daily laser therapy over 7 or 14 days transcutaneously starting 24 h from surgery. Incident energy density dosages of 31.5 and 94.5 Jcm-2 were applied during the period of the tibia wound healing investigated. Light microscopic examination of histological sections of the injured area and quantification of the newly-formed blood vessels were undertaken. Low-level energy treatment accelerated the deposition of bone matrix and histological characteristics compatible with an active recovery of the injured tissue. He-Ne laser therapy significantly increased the number of blood vessels after 7 days irradiation at an energy density of 94.5 Jcm-2, but significantly decreased the number of vessels in the 14-day irradiated tibiae, independent of the dosage. These effects were attributed to laser treatment, since no significant increase in blood vessel number was detected between 8 and 15 non-irradiated control tibiae. Molecular mechanisms involved in low-level laser therapy of angiogenesis in post-traumatic bone regeneration needs further investigation.

The use of NaOCl in combination with CHX produces cytotoxic product

Objectives: The combination of sodium hypochlorite (NaOCl) and chlorhexidine (CHX) yields a precipitate potentially toxic (PPT). The aim of this study was to evaluate the tissue response to implanted polyethylene tubes filled with PPT-soaked fibrin sponge. Methods: Forty rats received four polyethylene tubes each; each tube was filled with fibrin sponge soaked by 2.5 % NaOCl, 2.0 % CHX, PPT (2.5 % NaOCl plus 2.0 % CHX), or not soaked (control). The observation time points were 7, 15, 30, 60, and 90 days. At each time point, eight animals were killed, and the tubes and surrounding tissues were removed, fixed, and prepared for light microscopic analysis by performing glycol methacrylate embedding, serial cutting into 3-μm sections, and hematoxylin-eosin staining. Qualitative and quantitative evaluations of the reactions were performed. Results were statistically analyzed by Kruskal-Wallis test (p < 0.05). Results: All chemical solutions caused moderate reactions at 7 days. On day 30, PPT group was more cytotoxic than the control group and the CHX group (p < 0.05). On days 15 and 60, PPT group was more cytotoxic than the control group (p < 0.05). On day 90, there was no statistically significant difference between the different groups. Conclusion: PPT is more cytotoxic than NaOCl and CHX alone, particularly in the short term. Clinical significance: Protocols which suggest the use of CHX and NaOCl must be revised because this mixture produces cytotoxic product. © 2013 Springer-Verlag Berlin Heidelberg.

Morphologic and biomechanical changes of thoracic and abdominal aorta in a rat model of cigarette smoke exposure

Background: Smoking is the most relevant environmental factor that affects the development of aortic aneurysm. Smokers have elevated levels of elastase activity in the arterial wall, which leads to weakening of the aorta. The aim of this study was to verify whether cigarette smoke exposure itself is capable of altering the aortic wall. Methods: Forty-eight Wistar rats were divided into 2-, 4-, and 6-month experimental periods and into 2 groups: smokers (submitted to smoke exposure at a rate of 40 cigarettes/day) and nonsmokers. At the end of the experimental periods, the aortas were removed and cross-sectioned to obtain histologic specimens for light microscopic and morphometric analyses. The remaining longitudinal segments were stretched to rupture and mechanical parameters were determined. Results: A degenerative process (i.e., a reduction in elastic fibers, the loss of lamellar arrangement, and a reduction of smooth muscle cells) was observed, and this effect was proportional in intensity to the period of tobacco exposure. We observed a progressive reduction in the yield point of the thoracic aorta over time (P < 0.05). There was a decrease in stiffness (P < 0.05) and in failure load (P < 0.05) at 6 months in the abdominal aorta of rats in the smoking group. Conclusions: Chronic exposure to tobacco smoke can affect the mechanical properties of the aorta and can also provoke substantial structural changes of the arterial wall. © 2013 Elsevier Inc. All rights reserved.

Avaliação histológica do processo de reparação do tecido ósseo do rato frente à aplicação de um adesivo à base de cianoacrilato

This research objective was to verify the tissues reactions obtained, in surgical experimental bone defect, after the etil cyanoacrylate chemical adhesive application in rats parietal bones. 1t was investigated the acceptance or no acceptance of the tissues surround the bone defect in which was placed the etil cyanoacrylate and the control bone defect. The specimens were submitted to light microscopic analysis. 24 adults rats were used (Rattus norvergicus, albinus, Wistar) with the average weight to 300 gr. and were divided in two groups: treated and control. After the bone defects were realized, etil cyanoacrylate was placed over one of the defects at the parietal bone (treated group). On the other defect nothing was placed and it was used like control group. After 24 hours, 3,5,7,30 and sixty days after surgery, for animals of each group were sacrificed. The bone samples were removed and fixed in 10% formalin during 72 hours and analyzed using light microscopy. The results showed that the acute inflammatory responses was more observed at lhe treated group than the control group. The chemical adhesive was observed until 60 days follow the surgery and the tissues around it present normal appearance. The chemical adhesive, etil cyanaocrylate, did not impede at the bone repair process

Organization of Sporulated Oocysts of Eimeria funduli in the Gulf Killifish, Fundulus grandis

Oocysts of Eimeria funduli were studied by transmission electron microscopy in naturally-infected livers of the Gulf killifish, Fundulus grandis. Tissues were cryo-processed because membranous structures in the oocyst appear to hinder routine fixation and embedment. The oocyst wall (about 25 nm thick) was adjacent to the host cell and consisted of an outer membrane that limited the host cell cytoplasm and an inner membrane separated from the outer membrane by a narrow space. In some specimens, dense material was applied to the inner face of the inner membrane. Individual sporocysts were surrounded by a membranous "veil" (about 25 nm thick) that consisted of two unit membranes. Sporopodia, projections of the sporocyst wall, supported the veil. The sporocyst wall (130-150 nm thick) consisted of two layers, a thin electron-lucent outer layer (about 10 nm thick) and a thick electron dense inner layer (about 130 nm thick). Depending on the plane of section, the inner layer had transverse striations with periods of 3 to 4 nm or 12 to 15 nm. A narrow fissure, broadest at the anterior pole of the sporocyst, extended about one-third the length of the sporocyst wall. The posterior pole of the sporocyst was characterized by a bulbous swelling. Although this swelling resembled a Stieda body in light microscopic preparations, ultrastructurally, the swelling was a knoblike thickening in the sporocyst wall and did not plug a gap in this wall

Nine New Species of Bugula Oken (Bryozoa: Cheilostomata) in Brazilian Shallow Waters

Background: Bugula is a speciose genus of marine bryozoans, represented by both endemic and cosmopolitan species distributed in tropical and temperate waters and important to marine biologists because of the occurrence of many species in harbor and fouling communities, therefore as potential invaders. The southeastern Brazilian coast in the southern Atlantic hosts the highest known diversity of the genus, a status intimately associated with the intensity of collecting efforts. Methodology: Morphological data based on the examination of living specimens, scanning electron and light microscopic images, and morphometric analyses were used to assess the diversity of Bugula along the coastal areas of southern, northeastern, and southeastern Brazil. In this study, morphological species boundaries were based mainly on avicularian characters. For two morphologically very similar species, boundaries are partially supported by 16 S rDNA molecular data. Results: Nine species are newly described from Brazil, as follows: Bugula bowiei n. sp. (= Bugula turrita sensu Marcus, 1937) from the southern, northeastern, and southeastern coasts; Bugula foliolata n. sp. (= Bugula flabellata sensu Marcus, 1938), Bugula guara n. sp., Bugula biota n. sp. and Bugula ingens n. sp from the southeastern coast; Bugula gnoma n. sp. and Bugula alba n. sp. from the northeastern coast; Bugula rochae n. sp. (= Bugula uniserialis sensu Marcus, 1937) from the southern coast; and Bugula migottoi n. sp., from the southeastern and southern coasts. Conclusion: The results contribute to the morphological characterization and the knowledge of the species richness of the genus in the southwestern Atlantic (i.e., Brazil), through the description of new species in poorly sampled areas and also on the southeastern coast of that country. Additionally, the taxonomic status of the Brazilian specimens attributed to B. flabellata, B. turrita and B. uniserialis are clarified by detailed studies on zooidal and avicularia morphology.