Histogical and ultrastructural aspects of the brindley's glands of pantrongylus megistus (Burmeister, 1835) (Hemiptera: Reduviidae)

The Brindley's glands of Panstrongylus megistus were studied under the antomic, histologic and ultrastructural point of view. These glands located in the insect's methatorax are paired and have an opening near the third parir of the feet. Beside this aperture, ther are evaporation areas. Shape, sixe and aspect of the gland vary according to the feeding status. The glands are composed by a tubular part corresponding to the duct and a sack-like portion corrsponding to the secretory part. By electron microscopy we observed that the basal part of the epithelium has many interdigitations associated with mitochondria. On the apical surface where epicuticular foldings are located an electonlucent space is often seen. The glands are composed of the following elements: 1) superficial epithelial cells, located just below the apical surface foldings; 2) secretory cells; which are long and have an intracellular canalicule which changes according to the functional state of the cell; 3) a collecting duct to the secretory cells and covered with an epicuticle, reaching up to the gland's lumen; and 4) cells around the duct.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae). I. Development during the 3rd larval instar

The ultrastructure and distribution of gonial and somatic cells in the ovary of Dermatobia hominis was studied during the 3rd larval instar. In larvae weighing between 400 and 500 mg, the ovary is partially divided into basal and apical regions by oblong somatic cells that penetrate from the periphery; these cells show ovoid nucleus and cytoplasm full of microtubules. In both regions, gonial cells with regular outlines, large nucleus and low electron-density cytoplasm are scattered among the interstitial somatic cells. These later cells have small nucleus and electrodense cytoplasm. Clear somatic cells with small nucleus and cytoplasm of very low electron-density are restrict to the apical region of the gonad. Degenerating interstitial somatic cells are seen in the basal portion close to the ovary peduncle. During all this larval period the morphological features of the ovary remain almost the same. At the end of the period there is a gradual deposition of glycogen in the cytoplasm of the somatic cells, increase in the number and density of their mitochondria plus nuclear modification as membrane wrinkling and chromatin condensation in masses.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae). II. Origin of the tunica propria in ovarioles

Ovaries up to the 8th day pupae of Dermatobia hominis were studied by transmission electron microscopy. Ovarioles were recognized in ovaries of 4-day old pre-pupae, surrounded by a thin tunica propria of acellular fibrilar material similar in structure to the internal portion of the external tunica of the ovary. There is continuity of the tunica propria and the ovarian tunica, indicating that the former structure originates from the tunica externa. In 5 to 7-day pupae the interstitial somatic cells from the apical region of the ovary, close to the ovarioles, show delicate filamentous material inside of their rough endoplasmic reticulum cisternae; similar material is seem among these cells. Our observations suggest that interstitial somatic cells do not originate the tunica propria but contribute to its final composition.

Normal kinetics of intestinal glucose absorption in the absence of GLUT2: evidence for a transport pathway requiring glucose phosphorylation and transfer into the endoplasmic reticulum.

Glucose is absorbed through the intestine by a transepithelial transport system initiated at the apical membrane by the cotransporter SGLT-1; intracellular glucose is then assumed to diffuse across the basolateral membrane through GLUT2. Here, we evaluated the impact of GLUT2 gene inactivation on this transepithelial transport process. We report that the kinetics of transepithelial glucose transport, as assessed in oral glucose tolerance tests, was identical in the presence or absence of GLUT2; that the transport was transcellular because it could be inhibited by the SGLT-1 inhibitor phlorizin, and that it could not be explained by overexpression of another known glucose transporter. By using an isolated intestine perfusion system, we demonstrated that the rate of transepithelial transport was similar in control and GLUT2(-/-) intestine and that it was increased to the same extent by cAMP in both situations. However, in the absence, but not in the presence, of GLUT2, the transport was inhibited dose-dependently by the glucose-6-phosphate translocase inhibitor S4048. Furthermore, whereas transport of [(14)C]glucose proceeded with the same kinetics in control and GLUT2(-/-) intestine, [(14)C]3-O-methylglucose was transported in intestine of control but not of mutant mice. Together our data demonstrate the existence of a transepithelial glucose transport system in GLUT2(-/-) intestine that requires glucose phosphorylation and transfer of glucose-6-phosphate into the endoplasmic reticulum. Glucose may then be released out of the cells by a membrane traffic-based pathway similar to the one we previously described in GLUT2-null hepatocytes.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: Cuterebridae): III. Gonial cell degeneration

We studied the ultrastructural aspects of pre-pupae and pupae ovaries of Dermatobia hominis. Physiological degeneration of gonial cells was observed: (a) after the ovarioles differentiation, in the oogonia residing in the apical region of the ovary; (b) at the beginning of vitellogenesis, in the cystoblasts close to the terminal filament. The significance of gonial cell degeneration was correlated with the physiological changes wich occur in the ovary during development.

Gundlachia dutrae: n. sp. from northwest Brazil (Mollusca: Basommatophora: Ancylidae)

A new species of Gundlachia, Gundlachia dutrae is described from northwest Brazil. It is distinguishable from other congenerie species by characteristics of the shell, radula and internal organs. Shell relatively high. Aperture near-circular; periostracum dark brown without periostracal hairs. Apex slightly inclined to the right, projected but not hooked, with an apical depression surrounted by a sculpture of well-marked irregular punctations. Shell surface with prominent radial sculpture. No septate specimens were observed. Ratios (n= 59): shell width/shell lenght = 0,66- 0,79 (mean 0,73); shell height/shell length = 0,32- 0,45 ( mean 0,37); shell height/shell width = 0,43- 0,63 (mean 0,51). Body of normal ancylid type; mantle pigmentation dark brown or black, concentrated along the mantle collar. The dorsal surface of the right anterior muscle is elongated and medially constricted. The left anterior and the posterior muscles are almost elliptical. Adhesive area is V-shaped. Pseudobranch unpigmented bearing a very small and thin dorsal lobe. Ovotestis with more than 25 unbranched diverticula. Ovispermiduct with seminal vesicle rather developed. Elongated nidamental gland continous with the glandular wall of the uterus. Nidamental gland appendix ending into a bulbous swelling Spermathecal body almost rounded. Well-developed prostate with five long diverticula. Ejaculatory complex with long glandular flagellum, without a penis or true ultra-penis. "Penis sheath" developed. "Ultra-penis" projected as a tube inside the lumem of prepuce, with a slit between "ultra-penis" and "penis sheath". Rachidian tooth tetracuspid, with two median cusps assymmetrical and aculeated. Lateral teeth tricuspid, with a reduced endocon and a prominent mesocon. A well marked gap occurs between meso and ectocon. Marginal teeth similar to lateral ones. Jaw T-shaped, with about 28 dorsal plates.

Aspects of immunity for the AMA-1 family of molecules in humans and non-human primates malarias

The apical membrane antigen (AMA-1) family of malaria merozoite proteins is characterised by a high degree of inter-species conservation. Evidence that the protein (PK66/AMA-1) from the simian parasite Plasmodium knowlesi was protective in rhesus monkeys suggested that the 83kDa P. falciparum equivalent (PF83/AMA-1) should be investigated for protective effects in humans. Here we briefly review pertinent comparative data, and describe the use of an eukaryotic full length recombinant PF83/AMA-1 molecule to develop a sensitive ELISA for the determination of serological responses in endemic populations. The assay has revealed surprisingly high levels of humoral response to this quantitatively minor antigen. We also show that PK66/AMA-1 inhibitory mAb's are active against merozoites subsequent to release from schizont-infected red cells, further implicating AMA-1 molecules in red cell invasion.

International Society of Urological Pathology (ISUP) Consensus Conference on Handling and Staging of Radical Prostatectomy Specimens. Working group 5: surgical margins.

The 2009 International Society of Urological Pathology Consensus Conference in Boston, made recommendations regarding the standardization of pathology reporting of radical prostatectomy specimens. Issues relating to surgical margin assessment were coordinated by working group 5. Pathologists agreed that tumor extending close to the 'capsular' margin, yet not to it, should be reported as a negative margin, and that locations of positive margins should be indicated as either posterior, posterolateral, lateral, anterior at the prostatic apex, mid-prostate or base. Other items of consensus included specifying the extent of any positive margin as millimeters of involvement; tumor in skeletal muscle at the apical perpendicular margin section, in the absence of accompanying benign glands, to be considered organ confined; and that proximal and distal margins be uniformly referred to as bladder neck and prostatic apex, respectively. Grading of tumor at positive margins was to be left to the discretion of the reporting pathologists. There was no consensus as to how the surgical margin should be regarded when tumor is present at the inked edge of the tissue, in the absence of transected benign glands at the apical margin. Pathologists also did not achieve agreement on the reporting approach to benign prostatic glands at an inked surgical margin in which no carcinoma is present.

Unexpected left ventricular free-wall rupture following an aortic catheter-valve implantation.

Our experience with the Sapien trans-apical aortic valve (Edwards Lifesciences Inc., Irvine, CA, USA) has been straightforward without per-procedural mortality except in 1/16 consecutive cases who developed non-apical haemorrhage early after valve implantation. We describe the case of an 84-year-old female carrying a very high operative risk (logistic EuroScore of 44%), who underwent a trans-apical stent-valve implantation for severe and symptomatic aortic valve stenosis (23 mm). Due to massive blood loss, an emergency sternotomy and cannulation for cardiopulmonary bypass resuscitation were necessary to treat (without success) an unusual and unexpected subaortic left ventricular free-wall rupture that occurred few minutes after the stent-valve positioning and implantation. To the best of our knowledge, this is the first described case of a left ventricular free-wall rupture occurring after an otherwise non-complicated standard catheter-based aortic valve replacement.

Wnt secretion and gradient formation.

Concentration gradients formed by the lipid-modified morphogens of the Wnt family are known for their pivotal roles during embryogenesis and adult tissue homeostasis. Wnt morphogens are also implicated in a variety of human diseases, especially cancer. Therefore, the signaling cascades triggered by Wnts have received considerable attention during recent decades. However, how Wnts are secreted and how concentration gradients are formed remains poorly understood. The use of model organisms such as Drosophila melanogaster has provided important advances in this area. For instance, we have previously shown that the lipid raft-associated reggie/flotillin proteins influence Wnt secretion and spreading in Drosophila. Our work supports the notion that producing cells secrete Wnt molecules in at least two pools: a poorly diffusible one and a reggie/flotillin-dependent highly diffusible pool which allows morphogen spreading over long distances away from its source of production. Here we revise the current views of Wnt secretion and spreading, and propose two models for the role of the reggie/flotillin proteins in these processes: (i) reggies/flotillins regulate the basolateral endocytosis of the poorly diffusible, membrane-bound Wnt pool, which is then sorted and secreted to apical compartments for long-range diffusion, and (ii) lipid rafts organized by reggies/flotillins serve as "dating points" where extracellular Wnt transiently interacts with lipoprotein receptors to allow its capture and further spreading via lipoprotein particles. We further discuss these processes in the context of human breast cancer. A better understanding of these phenomena may be relevant for identification of novel drug targets and therapeutic strategies.

Ectopic expression of the serine protease inhibitor PI9 modulates death receptor-mediated apoptosis.

Apoptosis is a highly controlled process, whose triggering is associated with the activation of caspases. Apoptosis can be induced via a subgroup of the tumor necrosis factor (TNF) receptor superfamily, which recruit and activate pro-caspase-8 and -10. Regulation of apoptosis is achieved by several inhibitors, including c-FLICE-inhibitory protein, which prevents apoptosis by inhibiting the pro-apoptotic activation of upstream caspases. Here we show that the human intracellular serine protease inhibitor (serpin), protease inhibitor 9 (PI9), inhibits TNF-, TNF-related apoptosis-inducing ligand- and Fas ligand-mediated apoptosis in certain TNF-sensitive cell lines. The reactive center P1 residue of PI9 was required for this inhibition since PI9 harboring a Glu --> Ala mutation in its reactive center failed to impair death receptor-induced cell death. This suggests a classical serpin-protease interaction. Indeed, PI9 inhibited apoptotic death by directly interacting with the intermediate active forms of caspase-8 and -10. This indicates that PI9 can regulate pro-apoptotic apical caspases.

Triatoma jurbergi sp. n. do Norte do Estado do Mato Grosso, Brasil (Hemiptera, Reduviidae, Triatominae) com uma Atualização das Sinonímias e outros Táxons

Triatoma jurbergi n. sp. is described based on nine specimens of both sexes deposited in the Rodolfo Carcavallo Collection in the Oswaldo Cruz Institute Entomological Collection. The new species can be separated from the closely related Triatoma guazu Lent & Wygodzinsky, 1979 by several characters. The most important are longer anteocular region; thin and pointed juga; the shape of the eyes without concavity in the posterior edge; much longer second rostral segment, passing the posterior edge of eye; the absence of a ventral longitudinal depression on the abdomen; the general color redish, brown and orange and the male genitalia, mainly in the vesica lightly chitinized and smaller, the phallosome with apical projection and the pointed apex of the endosome process.

Polarity of endogenous and exogenous glycosyl-phosphatidylinositol-anchored membrane proteins in Madin-Darby canine kidney cells.

Madin-Darby canine kidney cells (MDCK) were transfected with a cDNA encoding the glycosyl-phosphatidylinositol (GPI)-anchored protein mouse Thy-1 in order to study the steady-state surface distribution of exogenous and endogenous GPI-linked proteins. Immunofluorescence of transfected cells grown on collagen-coated coverslips showed that expression of Thy-1 was variable throughout the epithelium, with some cells expressing large amounts of Thy-1 adjacent to very faintly staining cells. Selective surface iodination of cells grown on collagen-coated or uncoated transwell filters followed by immunoprecipitation of Thy-1 demonstrated that all the Thy-1 was present exclusively in the apical plasma membrane. Although cells grown on uncoated filters had much smaller amounts of Thy-1, it was consistently localized on the apical surfaces. Immunofluorescent localization of Thy-1 on 1 micron frozen sections of filter-grown cells demonstrated that all the Thy-1 was on the apical surface and there was no detectable intracellular pool. Phosphatidylinositol-specific phospholipase C digestion of intact iodinated monolayers released Thy-1 only into the apical medium, indicating that Thy-1 was processed normally in transfected cells and was anchored by a GPI-tail. In agreement with previous findings, endogenous GPI-linked proteins were found only on the apical plasma membrane. These results suggest that there is a common mechanism for sorting and targeting of GPI-linked proteins in polarized epithelial cells.

Glut9 is a major regulator of urate homeostasis and its genetic inactivation induces hyperuricosuria and urate nephropathy.

Elevated plasma urate levels are associated with metabolic, cardiovascular, and renal diseases. Urate may also form crystals, which can be deposited in joints causing gout and in kidney tubules inducing nephrolithiasis. In mice, plasma urate levels are controlled by hepatic breakdown, as well as, by incompletely understood renal processes of reabsorption and secretion. Here, we investigated the role of the recently identified urate transporter, Glut9, in the physiological control of urate homeostasis using mice with systemic or liver-specific inactivation of the Glut9 gene. We show that Glut9 is expressed in the basolateral membrane of hepatocytes and in both apical and basolateral membranes of the distal nephron. Mice with systemic knockout of Glut9 display moderate hyperuricemia, massive hyperuricosuria, and an early-onset nephropathy, characterized by obstructive lithiasis, tubulointerstitial inflammation, and progressive inflammatory fibrosis of the cortex, as well as, mild renal insufficiency. In contrast, liver-specific inactivation of the Glut9 gene in adult mice leads to severe hyperuricemia and hyperuricosuria, in the absence of urate nephropathy or any structural abnormality of the kidney. Together, our data show that Glut9 plays a major role in urate homeostasis by its dual role in urate handling in the kidney and uptake in the liver.

Scolex Morphology of Proteocephalid Cestodes Parasites of Neotropical Freshwater Fishes

The morphology of the scolices and metascolices of 29 proteocephalid species, parasites of freshwater fishes from Brazil was compared by using scanning electron microscope, light and laser microscopy. The following species were evaluated: Proteocephalus vazzolerae, P. piramutab, Robertiella paranaensis, Travassiella avitellina, Monticellia loyolai, M. spinulifera, M. belavistensis, Houssayela sudobim, Zygobothrium megacephalum, Gibsoniela mandube, Cangatiella arandasi, Nomimoscolex sudobim, N. lopesi, N. admonticellia, N. piraeeba, N. pirarara, Harriscolex kaparari, Crepidobothrium eirasi, Spatulifer rugosa, Megathylacus brooksi, Choanoscolex abscisus, Amphoteromorphus peniculus, A. piraeeba, Paramonticellia itaipuensis, Peltidocotyle rugosa, Othinoscolex lenha, Rudolphiella rugata, R. piranabu, Jauella glandicephalus. Some features as overall shape of scolex, suckers, apical sucker, frontal glands and several kinds of metascolex were analyzed. The taxonomic importance of scolex and metascolex is discussed.