Characterization of oral double balloon endoscopy in the dog.

Exploration of the canine small intestine using conventional endoscopy is restricted to the duodenum and/or the ileum. Double balloon endoscopy (DBE) is a 'push and pull' technique that has been described in humans and permits a complete exploration of the small intestine. In this study, oral DBE was performed on 12 healthy dogs (10-34kg) to characterize for the first time the efficiency, exploration dynamics and safety of the technique. DBE was successful in 83% of dogs; the average estimated insertion depth of the endoscope was 287±36cm, and the average duration of the exploration was 84±8min. No complications or relevant adverse clinical effects were observed, and there was no indication of post-procedure pancreatitis based on serology of two specific markers of pancreatitis (amylase and lipase) and the immediate nonspecific inflammatory mediator C-reactive protein. The study showed that oral DBE is viable and safe in the dog, allowing for the diagnosis and treatment of gastrointestinal diseases deep in the small intestine to an extent that has not previously been possible using conventional endoscopy.

Anatomical study of fibrous structures of the medial aspect of the canine elbow joint.

Forces exerted by fibrous structures on the medial aspect of the canine elbow joint have been reported to be involved in elbow pathology. The purpose of this paper is to assess the relationships of the ligamentous and muscular structures of this region of the canine elbow joint, and how they relate to the medial coronoid process. Six cadavers of adult German shepherd cross-breed dogs were used in this study to make intra-articular and vascular injections of this region. Some joints were dissected and some were frozen to saw sagittal or dorsal cryosections to assess the relationships of the myotendinous structures. The brachialis muscle tendon passed through the division of the bicipital tendon of insertion which formed a fibrous tendon sheath that was reinforced by the oblique ligament. The biceps' brachii's main insertion is the radial tuberosity where it inserts along with the cranial branch of the oblique ligament and the cranial branch of the medial collateral ligament. Rotational and compression forces exerted by the insertion of the biceps brachii-brachialis tendon complex onto the ulna might influence medial coronoid disease. Therefore, sectioning these tendons could be considered as a treatment for medial coronoid disease.

Morphometric study of the layers of the canine small intestine at five sampling sites.

The histology of the canine intestine has not been accurately defined. To establish the precise thickness of its different layers, whole wall samples of the small intestine were removed from 41 cadavers at five standardised sampling sites (duodenum, proximal jejunum, distal jejunum, proximal ileum and distal ileum). The total thickness was estimated by morphometry, as was the thickness of the mucosa, muscularis mucosae, submucosa and muscularis externa. In addition, the size of the lymphoid aggregates in the submucosa and the thickness of the circular and longitudinal layers within both the muscularis mucosae and the muscularis externa were estimated. The total intestinal thickness depended very much upon the thickness of the mucosa and submucosa. The mucosa decreased progressively from proximal to distal parts of the small intestine (47% reduction). The thickness of the submucosa, however, changed little from the duodenum to the distal jejunum, but increased significantly in the ileum; this change was positively correlated with the amount of lymphoid tissue. Sex influenced the thickness of the intestinal wall, with males displaying higher thickness values along the small intestine. Conversely, no correlation between bodyweight and intestinal thickness was found for any of the five sampling sites. This study gives absolute and relative values for the thickness of the layers of the dog intestine which might help in the diagnosis of small intestinal pathology from postmortem samples and/or endoscopic biopsies.

Morphometry of the canine intestine with reference to the use of double balloon endoscopy.

Double balloon endoscopy (DBE) enables the diagnosis and treatment of small intestinal disease. The dog is a potential animal model for DBE training and also a candidate for the clinical application of this technique. However, the anatomy of the canine small intestine may limit the use of DBE by restricting the push-and-pull manoeuvres required for DBE. To establish what these limitations are, the length and width of different portions of the small intestine as well as the mesenteric dependences were measured in 55 dogs. Several external parameters related to body size plus the age, weight, sex and breed were recorded. Potential restriction of the progress of the enteroscope due to narrow intestinal diameter is expected in the jejunum of mid- and small size dogs.

N-acetylcysteine inhibits TNF-alpha, sTNFR, and TGF-beta1 release by alveolar macrophages in idiopathic pulmonary fibrosis in vitro.

BACKGROUND: N-acetylcysteine (NAC) can act as an antioxidant. NAC slows the rate of decline of lung function in idiopathic pulmonary fibrosis (IPF) patients concurrently treated with prednisone and azathioprine. OBJECTIVE: In this study we investigated the effect of NAC on the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12 (p70), IL-18, transforming growth factor (TGF)-beta1, and the soluble TNF receptors (sTNFR1 and sTNFR2) by alveolar macrophages (AM) in IPF patients. DESIGN: AMs were harvested by bronchoalveolar lavage (BAL) from 16 IPF patients and were cultured for 24 h with RPMI medium alone, or with lipopolysaccharide (LPS) (100 ng/ml), in the presence or absence of NAC at various concentrations. RESULTS: NAC suppressed the production of TNF-alpha, its soluble receptors, and TGF-beta1 by AMs in a dose-dependent manner. At the highest concentration of NAC (10 mM), the spontaneous or LPS-stimulated production ofTNF-alpha, sTNFR1, sTNFR2, and TGF-beta1 were significantly reduced. The LPS-stimulated IL-1beta production was also suppressed by 10 mM NAC. CONCLUSIONS: NAC has the potential to down-regulate the production of TNF-alpha and their soluble receptors, as well as TGF-beta1 and LPS-stimulated IL-1beta, by AM in IPF in vitro. NAC may have anti-inflammatory and anti-fibrotic effects.

Thalidomide reduces IL-18, IL-8 and TNF-alpha release from alveolar macrophages in interstitial lung disease.

Thalidomide exhibits diverse actions of anti-inflammation, immunomodulation and anti-angiogenesis. The efficacy of thalidomide treatment in sarcoidosis with lupus pernio is thought to be due to inhibition of tumour necrosis factor (TNF)-alpha. The mechanisms that underlie the properties of thalidomide are still unclear in interstitial lung disease. The current authors investigated the potential inhibitory effects of thalidomide at concentrations of 0.1, 0.01 and 0.001 mM on the production of transforming growth factor-beta, TNF-alpha, interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12p70, IL-12p40 and IL-18 by alveolar macrophages from bronchoalveolar lavage in patients with sarcoidosis (n = 8), hypersensitivity pneumonitis (HP; n = 8) and idiopathic pulmonary fibrosis (IPF; n = 12). In sarcoidosis and HP patients, thalidomide induced a dose-dependent, partial suppression of lipopolysacchride (LPS)-stimulated TNF-alpha, IL-12p40 and IL-18 release. At the highest thalidomide concentration (0.1 mM), LPS-stimulated IL-8 production was also suppressed. In IPF patients, although spontaneous production of TNF-alpha, IL-12p40, IL-18 and IL-8 was lower than in sarcoidosis and HP patients, with LPS stimulation the cytokines were significantly elevated and also partially inhibited by thalidomide. In conclusion, thalidomide has the potential to improve the therapeutic regimens for sarcoidosis, hypersensitivity pneumonitis and idiopathic pulmonary fibrosis by reducing tumour necrosis factor-alpha, interleukin-12p40, interleukin-18 and interleukin-8 production.

Immunocytochemical localization of metabotropic (mGluR2/3 and mGluR4a) and ionotropic (GluR2/3) glutamate receptors in adrenal medullary ganglion cells.

The localization of metabotropic glutamate receptors of groups II (mGluR2/3) and III (mGluR4a) and the subunits 2 and 3 of alfa-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) ionotropic glutamate receptors (GluR2/3) was investigated with immunocytochemical methods in the rat adrenal gland. MGluR2/3, mGluR4a and GluR2/3 immunoreactivities were observed in large-sized, centrally located type I adrenal medullary ganglion neurons. Furthermore, the small-sized type II adrenal ganglion neurons identified by their immunoreactivity to brain nitric oxide synthase (bNOS), also expressed mGluR2/3, mGluR4a and GluR2/3. These cells were disposed in the peripheral portion of the adrenal medulla. None of the type I neurons were positively labeled for bNOS. These morphological observations suggest that activation of glutamate receptors in ganglion neurons may be instrumental in the control of adrenal endocrine systems as well as blood regulation.

[Supernumerary nostril]. / Tercera ventana nasal.

Supernumerary nostril is an extremely rare congenital anomaly as a result of aberrant embryological development. Frequently it leads to a narrow cavity; in the end, opening into the nearest nasal cavity or just stops blindly after a short course. It can appear isolated or in association with other anomalies. Treatment is always surgical. We present a case of a triple nostril, associated to a congenital auricular hypoplasia and we also review the existing literature.

Tercera ventana nasal / Supernumerary nostril

La tercera ventana nasal es una anomalía congénita muy rara que resulta de un desarrollo embriológico aberrante. La mayor parte de las veces se continúa hacia atrás en una cavidad nasal que a su vez puede comunicar con la fosa natural más cercana o acabar en fondo de saco. Puede aparecer aislada o asociada a otras anomalías. El tratamiento es quirúrgico. Presentamos un caso de tercera ventana nasal con hipoplasia de pabellón auricular derecho asociada, revisando la literatura existente (AU)

Supernumerary nostril is an extremely rare congenital anomaly as a result of aberrant embryological development. Frequently it leads to a narrow cavity; in the end, opening into the nearest nasal cavity or just stops blindly after a short course. It can appear isolated or in association with other anomalies. Treatment is always surgical. We present a case of a triple nostril, associated to a congenital auricular hypoplasia and we also review the existing literature (AU)

Cell proliferation, nuclear ploidy, and EGFr and HER2/neu tyrosine kinase oncoproteins in infiltrating ductal breast carcinoma.

The purpose of this work was to study if the variations in the nuclear DNA content and the expression of EGFr and HER2/neu transmembrane oncoproteins were related and if this influences the rate of cell proliferation modifying, in each case, the potential aggressivity of the neoplasia. Thirty-four ductal breast carcinoma tissue samples of tumors of up to 2 cm in diameter were analyzed by flow cytometry. HER2/neu and EGFr were measured by immunohistochemical methods. Twenty cases were diploid (DNA index of 1) and 14 cases were aneuploid (DNA index other than 1). The expression of EGFr and HER2/neu was significantly higher in aneuploid tumors compared with diploid tumors. The cell proliferation rate was significantly higher in tumors with an aneuploid pattern. The expression of EGFr was associated with a higher rate of cell proliferation. The higher expression of EGFr and HER 2/neu oncoproteins in aneuploid tumors suggests that the increased proliferative activity of aneuploid carcinomas is influenced by the activity of such oncoproteins, which favors a more aggressive biological behavior.

Rat neonatal intestinal wall-free graft.

An experimental study was carried out in order to evaluate the regeneration capacity of the neonatal intestinal wall in ischaemia and its repercussion over organs of the immune system such as the spleen. We isolated a reservoir of small intestine in adult Sprague-Dawley rats that, after having been everted and placed at a subcutaneous level, was grafted with a free small intestine segment of neonatal Sprague-Dawley rats and analysed 3, 15 and 30 days after grafting. Samples obtained were stained with Martin's trichromic stain and studied at the light microscopic level. A total regeneration of the crypt architecture, formed by absorptive enterocytes, was observed in the interior of the reservoirs. A quantitative immunohistochemical study with monoclonal antibodies against CD4, CD8, MHC I and MHC II was carried out in the spleen of these animals. An additional immunohistochemical study was also performed in the small intestine reservoirs and spleen of transplanted animals using nitric oxide synthase (NOS) antibodies. Three days after transplantation NOS immunoreactive cells were observed in the reservoirs with transplanted neonatal small intestine. Antigenic stimulation produced, in the spleen red pulp of transplanted animals, a significant increase (P < 0.001) in the percentage of NOS, CD8 and MHC I immunoreactive cells in relation with values observed in control animals. These morphometric changes could be related with the stimulation that nitric oxide produces in the proliferation of the cytotoxic T cells.

Characterization of a family of Arabidopsis genes related to xyloglucan fucosyltransferase1.

To understand primary cell wall assembly in Arabidopsis, we have focused on identifying and characterizing enzymes involved in xyloglucan biosynthesis. Nine genes (AtFUT2-10) were identified that share between 47% and 62% amino acid similarity with the xyloglucan-specific fucosyltransferase AtFUT1. Reverse transcriptase-PCR analysis indicates that all these genes are expressed. Bioinformatic analysis predicts that these family members are fucosyltransferases, and we first hypothesized that some may also be involved in xyloglucan biosynthesis. AtFUT3, AtFUT4, and AtFUT5 were expressed in tobacco (Nicotiana tabacum L. cv BY2) suspension culture cells, and the resulting proteins did not transfer fucose (Fuc) from GDP-Fuc to tamarind xyloglucan. AtFUT3, AtFUT4, and AtFUT5 were overexpressed in Arabidopsis plants. Leaves of plants overexpressing AtFUT4 or AtFUT5 contained more Fuc than wild-type plants. Stems of plants overexpressing AtFUT4 or AtFUT5 contained more xylose, less arabinose, and less galactose than wild-type plants. We suggest that the AtFUT family is likely to include fucosyltransferases important for the synthesis of wall carbohydrates. A targeted analysis of isolated cell wall matrix components from plants altered in expression of these proteins will help determine their specificity and biological function.

Eficacia terapéutica del Nimesulide en el tratamiento de la dorso lumbalgia de origen traumático / Therapeutic effectiveness of the nimesulide in the treatment of the low back pain of traumatic origin

Se realizó un estudio abierto, no controlado, multicéntrico, realizado con intención de tratar, en 60 pacientes con dorsolumbalgia mecánica, en el cual evaluó la efectividad y seguridad de nimesulide 100 mg dos veces al día, administrado durante 15 días. El tratamiento produjo un alivio significativo en los escores de dolor entre el día de inicio del tratamiento y los 5 días, con un alivio adicional importante entre los días 5 y 10, sin alivios adicionales para el día 15. El tiempo medio para la desaparición total del dolor fue de 8,5 días. La limitación funcional fue evaluada el día 0 y el día 15, obteniéndose una mejoría significativa desde el punto de vista estadístico. Se reportaron efectos adversos en 4 de los pacientes, de los cuales sólo uno ameritó la suspensión del tratamiento

[Endoscopic surgery of nasopharyngeal angiofibroma by double embolization]. / Cirugía endoscópica del angiofibroma nasofaríngeo mediante doble embolización.

Juvenile nasopharyngeal angiofibroma is a source of controversy with respect to therapy because of the many therapeutic modalities that exist, such as classic surgical techniques and newer techniques like nasal endoscopic surgery, which have emerged with the advent of new and better diagnostic techniques. Endoscopic surgery is less aggressive and produces less morbidity, but its use often depends on the size and extension of the tumor. A clinical case is presented with the subsequent diagnostic steps and surgical treatment. Rigid endoscopy was used for double cynoacrylate embolization, by angiography before operation and intratumoral injection during surgery.

Cirugía endoscópica del angiofibroma nasofaríngeo mediante doble embolización / Endoscopic surgery of nasopharyngeal angiofibroma by double embolization

El angiofibroma nasofaríngeo juvenil es motivo de controversia respecto a la actitud terapéutica a tomar, debido a las variadas modalidades que existen en la actualidad, entre las que están las técnicas quirúrgicas clásicas y otras más novedosas como la cirugía endoscópica nasal, surgidas por el advenimiento de nuevas y mejores técnicas diagnósticas. La cirugía endoscópica es menos agresiva, con menor morbilidad, pero muchas veces supeditada al tamaño y extensión del tumor. Se presenta un caso clínico, con los pasos diagnósticos realizados y posterior cirugía. En la intervencíón se usa el endoscopio rígido, tras doble embolización: mediante angiografía, previo a la intervención y durante la cirugía, inyectando intratumoralmente cianocrilato (AU)

Juvenile nasopharyngeal angiofibroma is a source of controversy with respect to therapy because of the many therapeutic modalities that exist, such as classic surgical techniques and newer techniques like nasal endoscopic surgery, which have emerged with the advent of new and better diagnostic techniques. Endoscopic surgery is less aggressive and produces less morbidity, but its use often depends on the size and extension of the tumor. A clinical case is presented with the subsequent diagnostic steps and surgical treatment. Rigid endoscopy was used for double cynoacrylate embolization, by angiography before operation and intratumoral injection during surgery (AU)

Immunolocalization of the mGluR1b splice variant of the metabotropic glutamate receptor 1 at parallel fiber-Purkinje cell synapses in the rat cerebellar cortex.

Several metabotropic glutamate receptor (mGluR) subtypes have been identified in the cerebellar cortex that are targeted to different compartments in cerebellar cells. In this study, preembedding immunocytochemical methods for electron microscopy were used to investigate the subcellular distribution of the mGluR1b splice variant in the rat cerebellar cortex. Dendritic spines of Purkinje cells receiving parallel fiber synaptic terminals were immunoreactive for mGluR1b. With a preembedding immunogold method, approximately 25% of the mGluR1b immunolabeling was observed perisynaptically within 60 nm from the edge of the postsynaptic densities. Values of extrasynaptic gold particles beyond the first 60 nm were maintained at between 10 and 18% along the whole intracellular surface of the dendritic spine membranes of Purkinje cells. For comparison, the distribution of mGluR1a was studied. A predominant (approximately 37%) perisynaptic localization of mGluR1a was seen in dendritic spines of Purkinje cells, dropping the extrasynaptic labeling to 15% in the 60-120-nm bin from the edge of the postsynaptic specialization. Our results reveal that mGluR1b and mGluR1a are localized to the same subcellular compartments in Purkinje cells but that the densities of the perisynaptic and extrasynaptic pools were different for both isoforms. The compartmentalization of mGluR1b and mGluR1a might serve distinct requirements in cerebellar neurotransmission.

Transgenic plants of cassava (Manihot esculenta) with resistance to Basta obtained by Agrobacterium-mediated transformation.

Transgenic plants of cassava (Manihot esculenta) resistant to the herbicide Basta were obtained through Agrobacterium-mediated transformation. The plants also expressed the uidA gene and two were positive for PCR- and/or Southern-based detection of the nptII gene. Somatic-embryo-derived cotyledons were used as source of explants. A non-disarmed Agrobacterium strain (CIAT 1182) was used to transfer the genes of interest into cassava cultivar MPer183. Greenhouse tests of resistance to Basta (Hoechst) showed three plant lines with different levels of tolerance to the herbicide. Based on Southern tests of transgenesis, the transformation efficiency was 1%. The results constitute the first report of the bar gene conferring herbicide resistance to cassava plants.

Distribution of the group II metabotropic glutamate receptors (mGluR2/3) in the enteric nervous system of the rat.

We used affinity purified antisera specific for group II metabotropic glutamate receptors in combination with a pre-embedding immunocytochemical method for light microscopy to investigate the localization of mGluR2/3 receptors in the enteric nervous system of the rat small intestine. In the submucosal plexuses of the jejunum and ileum, strongly mGluR2/3 immunoreactive cells were distributed between the circular muscular layer and the muscularis mucosa. In addition, oval or rounded immunostained ganglion cells appeared located in the myenteric plexuses of both intestinal regions. Nerve fibers intensely stained were observed extending between adjacent myenteric ganglia. The localization of mGluR2/3 receptors in enteric neurons might have functional implications in the physiology and pathology of the gut.

Stoichiometric shifts in the common bean mitochondrial genome leading to male sterility and spontaneous reversion to fertility

The plant mitochondrial genome is characterized by a complex, multipartite structure. In cytoplasmic male-sterile (CMS) common bean, the sterility-inducing mitochondrial configuration maps as three autonomous DNA molecules, one containing the sterility-associated sequence pvs-or f 239. We constructed a physical map of the mitochondrial genome from the direct progenitors to the CMS cytoplasm and have shown that it maps as a single, circular master configuration. With long-exposure autoradiography of DNA gel blots and polymerase chain reaction analysis, we demonstrate that the three-molecule CMS-associated configuration was present at unusually low copy number within the progenitor genome and that the progenitor form was present substoichiometrically within the genome of the CMS line. Furthermore, upon spontaneous reversion to fertility, the progenitor genomic configuration as well as the molecule containing the pvs-or f 239 sterility-associated sequence were both maintained at substoichiometric levels within the revertant genome. In vitro mitochondrial incubation results demonstrated that the genomic shift of the pvs-or f 239-containing molecule to substoichiometric levels upon spontaneous reversion was a reversible phenomenon. Moreover, we demonstrate that substoichiometric forms, apparently silent with regard to gene expression, are transcriptionally and translationally active once amplified. Thus, copy number suppression may serve as an effective means of regulating gene expression in plant mitochondria.

A cytoplasmic male sterility-associated mitochondrial peptide in common bean is post-translationally regulated.

Cytoplasmic male sterility in the common bean plant is associated with a dominant mitochondrial mutation designated pvs-or f 239 (for Phaseolus vulgaris sterility sequence open reading frame 239). The sequence is transcribed in both vegetative and reproductive tissues, but the translation product, ORF239, is present only in reproductive tissues. We present evidence to support a model of post-translational regulation of ORF239 expression based on the following observations. In organello translation experiments using purified mitochondria from young seedlings demonstrated accumulation of ORF239 only when a protease inhibitor was included. Proteolytic activity against ORF239 was observed in mitochondrial extracts fractionating with the mitochondrial inner membrane. The DNA sequence encoding a serine-type protease, similar to the lon protease gene of Escherichia coli, was cloned from the Arabidopsis genome. The expression product of this sequence demonstrated proteolytic activity against ORF239 in vitro, with features resembling the activity detected in mitochondrial inner membrane preparations. Antibodies generated against the overexpressed Lon homolog reduced proteolytic activity against ORF239 when added to mitochondrial extracts. Our data suggest that ORF239 was undetected in vegetative tissue due to rapid turnover by at least one mitochondrial protease that acts against ORF239 post-translationally.