Expresión y rol del slpi en la inmunidad innata del ojo asociado a procesos inflamatorios e infecciosos oculares.

La investigación oftalmológica actual se focaliza en el estudio de los procesos cicatrizales normales y patológicos del globo ocular. Nuestro proyecto se focaliza en el estudio de los mecanismos fisiopatológicos oftalmológicos relacionados a la inmunidad innata y procesos inflamatorios e infecciosos oculares. Actualmente la terapia antimicrobiana y de cicatrización tisular pretende comprender y desarrollar sinergia entre los mecanismos involucrados en la inmunidad innata y cicatrizacion del huésped. En el área oftalmológica, son de crítica importancia el conocimiento y modulación de mencionados procesos oculares. Modulando y estimulando la presencia de los péptidos antimicrobianos y anti inflamatorios, se obtendría una prevención, modulación de los principales procesos fisiopatologicos oculares, permitiendo estudiar y aplicar una nueva área de tratamiento en las enfermedades oculares.De esta forma se podrá evitar secuelas devastadoras de dichas complicaciones que en un gran porcentaje llevan a la ceguera del paciente, asociado a la destrucción de tejidos específicos como la transparencia corneal o la perdida de neuronas retinales. Los péptidos antimicrobianos y anti-inflamatorios prometen ser un método terapéutico eficaz, natural, libre de efectos secundarios y adversos. Basados en estudios publicados (propios y de otros autores) realizados en animales, demuestran péptidos con capacidad antibacteriana e inmuno moduladora que se expresarían en tejidos oculares normales, actuando como agentes antimicrobianos de la inmunidad innata del ojo y así también como regulador de la actividad inflamatoria. Estos péptidos antimicrobianos – anti inflamatorios podrían utilizarse a futuro para el desarrollo en la prevención y tratamiento de procesos infecciosos oculares, en formulaciones simples o combinadas a otras sustancias antibióticas antimicrobianas en forma sinérgica. Secretory leukocyte protease inhibitor (SLPI), proteína de 12 kDa de peso molecular cuya función es inhibir proteasas de la matriz extracelular y modular la inmunidad del tejido involucrado. El objetivo es estudiar la presencia del péptido antimicrobiano SLPI en relación con el remodelado cicatrizal de la matriz extracelular de la cornea y otras estructuras intra oculares, como así también su expresión en relación a procesos Inflamatorios e infecciosos del ojo como agente antimicrobiano de la inmunidad innata del huésped.

Ultraestructura de los acinos sudoríparos de las glándulas pelvianas de Chaetophractus villosus (Mammalia, Dasypodidae)

The acini of pelvian glands of Chaetophractus villosus (Desmarest, 1804) consisted of an inner layer of secretory cells and an outer layer of myoepithelial cells. Secretory cells have numerous secretory vacuoles. The secretion is released by exocytosis. Myoepithelial cells have numerous myofilaments that occupy much of the cytoplasm. There is a third cell type with an extremely electron-lucent cytoplasm.

Estudos sobre thrombose cardiaca e endocardite parietal de origem não valvular

1.-Since the parietal endocarditis represents a chapter generally neglected, owing to the relative lack of cases, and somewhat confused because there various terms have been applied to a very same morbid condition, it justifies the work which previously we tried to accomplish, of nosographic classification. Taking into account the functional disturbances and the anatomical changes, all cases of parietal endocarditis referred to in the litterature were distributed by the following groups: A-Group-Valvulo-parietal endocarditis. 1st . type-Valvulo-parietal endocarditis per continuum. 2nd. type-Metastatic valvulo-parietal endocarditis. 3rd. type-Valvulo-parietal endocarditis of the mitral stenosis. B-Group-Genuine parietal endocarditis. a) with primary lesions in the myocardium. b) with primary lesions in the endocardium. 4th type-Fibrous chronic parietal endocarditis (B A Ü M L E R), « endocarditis parietalis simplex». 5th type-Septic acute parietal endocarditis (LESCHKE), «endocarditis parietalis septica». 6th type-Subacute parietal endocarditis (MAGARINOS TORRES), «endocarditis muralis lenta». 2.-Studying a group of 14 cases of fibrous endomyocarditis with formation of thrombi, and carrying together pathological and bacteriological examinations it has been found that some of such cases represent an infectious parietal endocarditis, sometimes post-puerperal, of subacute or slow course, the endocardic vegetations being contamined by pathogenic microörganisms of which the most frequent is the Diplococcus pneumoniae, in most cases of attenuated virulence. Along with the infectious parietal endocarditis, there occur arterial and venous thromboses (abdominal aorta, common illiac and femural arteries and external jugular veins). The case 5,120 is a typical one of this condition which we name subacute parietal endocarditis (endocarditis parietalis s. muralis lenta). 3.-The endocarditis muralis lenta encloses an affection reputed to be of rare occurrence, the «myocardite subaigüe primitive», of which JOSSERAND and GALLAVARDIN published in 1901 the first cases, and ROQUE and LEVY, another, in 1914. The «myocardite subaigüe primitive» was, wrongly, in our opinion, included by WALZER in the syndrome of myocardia of LAUBRY and WALZER, considering that, in the refered cases of JOSSERAND and GALLAVARDIN and in that of ROQUE and LEVY, there are described rather considerable inflammatory changes in the myocardium and endocardium. The designation «myocardia» was however especially created by LAUBRY and WALZER for the cases of heart failure in which the most careful aetiologic inquiries and the most minucious clinical examination were unable to explain, and in which, yet, the post-mortem examination did not reveal any anatomical change at all, it being forcible to admit, then, a primary functional change of the cardiac muscle fibre. This special cardiac condition is thoroughly exemplified in the observation that WALZER reproduces on pages 1 to 7 of his book. 4.-The clinical picture of the subacute parietal endocarditis is that of heart failure with oedemas, effusion in the serous cavities and passive chronic congestion of the lungs, liver, kideys and spleen associated, to that of an infectious disease of subacute course. The fever is rather transient oscillating around 99.5 F., being intersected with apyretic periods of irregular duration; it is not dependent on any evident extracardiac septic infection. In other cases the fever is slight, particularly in the final stage of the disease, when the heart failure is well established. The rule is to observe then, hypothermy. The cardiac-vascular signs consist of enlargement of the cardiac dullness, smoothing of the cardiac sounds, absence of organic murmurs and accentuated and persistent tachycardia up to a certain point independent of fever. The galloprhythm is present, in most cases. The signs of the pulmonary infarct are rather expressed by the aspect of the sputum, which is foamy and blood-streaked than by the classic signs. Cerebral embolism was a terminal accident on various cases. Yet, in some of them, along with the signs of septicemia and of cardiac insufficiency, occurred vascular, arterial (abdominal aorta, common illiac and femurals arteries) and venous (extern jugular veins) thromboses. 5. The autopsy revealed an inflammatory process located on the parietal endocardium, accompanied by abundant formation of ancient and recent thrombi, being the apex of the left ventricle, the junction of the anterior wall of the same ventricle, with the interventricular septum, and the right auricular appendage, the usual seats of the inflammatory changes. The region of the left branch of HIS’ bundle is spared. The other changes found consist of fibrosis of the myocardium (healed infarcts and circumscribed interstitial myocarditis), of recent visceral infarcts chiefly in lungs, spleen and brain, of recent or old infarcts in the kidneys (embolic nephrocirrhosis) and in the spleen, and of vascular thromboses (abdominal aorta, common illiacs and femurals arteries and external jugular veins), aside from hydrothorax, hydroperitoneum, cutaneous oedema, chronic passive congestion of the liver, lungs, spleen and kidneys and slight ictericia. 6. In the subacute parietal endocarditis the primary lesions sometimes locate themselves at the myocardium, depending on the ischemic necrosis associated to the arteriosclerosis of the coronariae arteries, or on an specific myocarditis. Other times, the absence of these conditions is suggestive of a primary attack to the parietal endocardium which is then the primary seat of the lesions. It matters little whatever may be the initial pathogenic mechanism; once injured the parietal endocardium and there being settled the infectious injury, the endocarditis develops with peculiar clinical and anatomical characters of remarkable uniformity, constituting an anatomo-clinical syndrome. 7.-The histologic sections show that recent lesions…

Reticulo-endotheliomatose maligna

A spindle-cell sarcoma (fig. 5) apparently originating from the dura (fig. 4) was found at the autopsy of a male, mulato, 17 years of age. The bones of the skull (occipital and both parietals) were penetrated and destroyed (fig. 1 and 2). The nervous tissue was not penetrated, the only change in the brain being a depressed area where the tumor was included. Metastatic nodules were found in the liver (fig. 3),hepatic lymphnodes (fig. 14), spleen (fig. 12) and suprarenal bodies (fig. 15). The structure, however, in all those different locations was that of a typical endothelioma (figs. 8, 11 and 13). The cells are of large and moderate size, of polyhedral form, with vesicular nuclei, diminutive nucleoli and clear cytoplasm. (Figs. 6 and 8). They are arranged about a central lumen which represents a rudimentary vessel (figs. 9 and 13). Other areas are composed of cells without concentric arrangement (figs. 4 and 10). In small areas, the colums of liver cells are marginated in one side by typical sinusoids, while in the other side tumor cells arranged about a narrow lumen are seen suggesting a pathological (neoplastic) sinusoid (figs. 7 and 9). The case is considered as a multiple diffuse endothelioma. The origin of the tumor is referred to the reticulo-endothelial apparatus of the liver, the spleen, the suprarenal bodies and the lymph nodes, the structure being rather uniform in those organs. In the dura, the endothelioma reproduces the structure and presents the general character of a fibroblastic sarcoma; in some places, however, the structure of endothelioma could be found (fig.6). It corresponds to the reticulo-endotheliomatosis maligna according to Puhr's grouping of progressive changes in the reticulo-endothelial apparatus which is a follows: 1. HYPERPLASTIC - 1. Mnnocytic leukemia. 2. a) Aleukemic reticulosis (Goldschmid and Isaac). b) Idiopathic sarcoma of skin (Kaposi). c) Cutaneous sarcoid (Spiegler). 3. Secretory reticulosis. a) Gaucher's disease. b) Generalized xanthomatosis. c) Spleno-hepatomegaly with lipoidic cells (Pick). II. BLASTOMATOSUS OR NEOPLASTIC - 1. Benign - a) Circumscribed tumors. a) Epulis sarcomatosa; b) Benign giant-cells sarcoma of the bone - marrow of long bones. b) Generalized brown tumors of osteitis fibrosa. 2. Malignant - a) Circumscribed haemangio - endothelioma (reticulo- endothelioma (maligum). of {liver, spleen, bone-marrow. b) Generalized haemangio-endotheliomatosis (reticulo-endotheliomatosis maligna) (Grabowski).

Insulin secretion is regulated by the glucose-dependent production of islet beta cell macrophage migration inhibitory factor.

Macrophage migration inhibitory factor (MIF), originally identified as a cytokine secreted by T lymphocytes, was found recently to be both a pituitary hormone and a mediator released by immune cells in response to glucocorticoid stimulation. We report here that the insulin-secreting beta cell of the islets of Langerhans expresses MIF and that its production is regulated by glucose in a time- and concentration-dependent manner. MIF and insulin colocalize by immunocytochemistry within the secretory granules of the pancreatic islet beta cells, and once released, MIF appears to regulate insulin release in an autocrine fashion. In perifusion studies performed with isolated rat islets, immunoneutralization of MIF reduced the first and second phase of the glucose-induced insulin secretion response by 39% and 31%, respectively. Conversely, exogenously added recombinant MIF was found to potentiate insulin release. Constitutive expression of MIF antisense RNA in the insulin-secreting INS-1 cell line inhibited MIF protein synthesis and decreased significantly glucose-induced insulin release. MIF is therefore a glucose-dependent, islet cell product that regulates insulin secretion in a positive manner and may play an important role in carbohydrate metabolism.

Transport of methotrexate by the in vitro isolated rabbit proximal tubule.

The tubular transport of [3H]methotrexate was studied in isolated nonperfused and perfused superficial proximal tubular segments of rabbit kidneys. Reabsorption represented only 5% of perfused methotrexate, and appeared to be mostly of passive nature inasmuch as it was not modified by reducing the temperature or by ouabain. Cellular accumulation in nonperfused segments and secretion in perfused tubules were highest in the S2 segment and lower in the S3 and S1 segments. Secretion against a bath-to-lumen concentration gradient was observed only in S2 segments (with a maximum methotrexate secretory rate of 478 +/- 48 fmol/mm.min and an apparent Km of transport of 363 +/- 32 microM), and was inhibited by probenecid and folate. The low capacity for methotrexate secretion may be explained by a low capacity of transport across the basolateral membrane of the proximal cell as methotrexate was accumulated only to a low extent in nonperfused tubules (tissue water to medium concentration ratio of 8.2 +/- 1 in S2 segments). During secretion a small amount of methotrexate was metabolized; the nature of the metabolite(s) remains to be defined.

Paper de la PAP en la resposta inflamatoria durant la pancreatitis aguda

Estudi elaborat a partir d’una estada al Institut national de la santé et de la recherche médicale, França, entre setembre i octubre del 2006. La PAP va ser identificada inicialment com una proteïna de secreció, que apareixia al suc pancreàtic després de la inducció d’una pancreatitis aguda experimental. Per la PAP s’ha suggerit diferents funcions, algunes no relacionades en aparença, però les més interessants en el camp de la pancreatitis són les activitats antiapoptótiques, mitogéniques i, especialment, antiinflamatóries. Per aprofundir en aquests aspectes de la PAP, s’ha emprat un model de ratolí PAP-/- per observar els efectes de la deleció del gen de la PAP durant la pancreatitis aguda.Per induir la pancreatitis es va fer servir el model de administració de ceruleina a dosi supra-màximes. En aquestes condicions es va observar que en els animals PAP-/-, la severitat del procés era menor. Els marcadors de necrosi pancreàtica, lipasa i amilasa, van presentar nivells menors en els animals PAP-/- que en els corresponents wild type. Per contra, el nombre de PMN infiltrats i la producció de citoquines pro-inflamatories va ser major en el pàncreas dels animals PAP-/-. La intensitat de la resposta inflamatòria observada suggereix que en condicions fisiològiques, el paper anti-inflamatori de la PAP es prou rellevant. Això ja s’havia suggerit en estudis in vitro, en els que es va demostrar que l’activitat antiinflamatòria de la PAP depenia de la via de transducció de senyal Jak/STAT/SOCS3. Aquesta hipòtesi s’ha comprovat in vivo, monitoritzant el nivell d’activació de STAT3 en els pàncreas dels animals després de la inducció de la pancreatitis.Tot plegat confirma que les funcions antiinflamatòries descrites in vitro per la PAP també es poden observar in vivo, de manera que la PAP sembla ser un agent important en la resposta de les cèl•lules pancreàtiques durant la pancreatitis aguda.

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Primary ciliary dyskinesia (PCD) is an autosomal recessive disease with an incidence estimated between 1:2,000 and 1:40,000. Ciliated epithelia line the airways, nasal and sinus cavities, Eustachian tube and fallopian tubes. Congenital abnormalities of ciliary structure and function impair mucociliary clearance. As a consequence, patients present with chronic sinopulmonary infections, recurrent glue ear and female subfertility. Similarities in the ultrastructure of respiratory cilia, nodal cilia and sperm result in patients with PCD also presenting with male infertility, abnormalities of left-right asymmetry (most commonly situs inversus totalis) and congenital heart disease. Early diagnosis is essential to ensure specialist management of the respiratory and otological complications of PCD. Diagnostic tests focus on analysis of ciliary function and electron microscopy structure. Analysis is technically difficult and labour intensive. It requires expertise for interpretation, restricting diagnosis to specialist centres. Management is currently based on the consensus of experts, and there is a pressing need for randomised clinical trials to inform treatment.

Nasal immunization of mice with human papillomavirus type 16 virus-like particles elicits neutralizing antibodies in mucosal secretions.

To specifically induce a mucosal antibody response to purified human papillomavirus type 16 (HPV16) virus-like particles (VLP), we immunized female BALB/c mice orally, intranasally, and/or parenterally and evaluated cholera toxin (CT) as a mucosal adjuvant. Anti-HPV16 VLP immunoglobulin G (IgG) and IgA titers in serum, saliva, and genital secretions were measured by enzyme-linked immunosorbent assay (ELISA). Systemic immunizations alone induced HPV16 VLP-specific IgG in serum and, to a lesser extent, in genital secretions but no secretory IgA. Oral immunization, even in the presence of CT, was inefficient. However, three nasal immunizations with 5 microgram of VLP given at weekly intervals to anesthetized mice induced high (>10(4)) and long-lasting (>15 weeks) titers of anti-HPV16 VLP antibodies in all samples, including IgA and IgG in saliva and genital secretions. CT enhanced the VLP-specific antibody response 10-fold in serum and to a lesser extent in saliva and genital secretions. Nasal immunization of conscious mice compared to anesthetized mice was inefficient and correlated with the absence of uptake of a marker into the lung. However, a 1-microgram VLP systemic priming followed by two 5-microgram VLP intranasal boosts in conscious mice induced both HPV16 VLP-specific IgG and IgA in secretions, although the titers were lower than in anesthetized mice given three intranasal immunizations. Antibodies in serum, saliva, and genital secretions of immunized mice were strongly neutralizing in vitro (50% neutralization with ELISA titers of 65 to 125). The mucosal and systemic/mucosal HPV16 VLP immunization protocols that induced significant titers of neutralizing IgG and secretory IgA in mucosal secretions in mice may be relevant to genital HPV VLP-based human vaccine trials.

Aspergillus fumigatus fungus ball in hospitalized patients with chronic pulmonary disease: usefulness of double immunodiffusion test as a screening procedure

Double immunodiffusion (DID) was used as a screening test for the diagnosis of aspergillosis. Three hundred and fifty patients were tested, all of them referred from a specialized chest disease hospital and without a definitive etiological diagnosis. When DID was positive addtional information such as clinical history and radiographic findings were requested and also surgical specimens were obtained whenever possible. Specific precipitin hamds for Aspergillus fumigatus antigen were found in 29 (8.3%) of 350 patients sera. Nineteen (65.5%) of the 29 patients with positive serology were recognized as having a fungus ball by X-rays signs in 17 or by pathological examination in 2 or by both in 8 patients. This two-year prospective study has shown that pulmonary aspergillos is a considerable problem among patiens admitted to a Chest Diseases Hospital, especially in those with pulmonary cavities or bronchiectasis.

A gene knockout approach in mice to identify glucose sensors controlling glucose homeostasis.

A key aspect of glucose homeostasis is the constant monitoring of blood glucose concentrations by specific glucose sensing units. These sensors, via stimulation of hormone secretion and activation of the autonomic nervous system (ANS), regulate tissue glucose uptake, utilization or production. The best described glucose detection system is that of the pancreatic beta-cells which controls insulin secretion. Secretion of other hormones, in particular glucagon, and activation of the ANS, are regulated by glucose through sensing mechanisms which are much less well characterized. Here I review some of the studies we have performed over the recent years on a mouse model of impaired glucose sensing generated by inactivation of the gene for the glucose transporter GLUT2. This transporter catalyzes glucose uptake by pancreatic beta-cells, the first step in the signaling cascade leading to glucose-stimulated insulin secretion. Inactivation of its gene leads to a loss of glucose sensing and impaired insulin secretion. Transgenic reexpression of the transporter in GLUT2/beta-cells restores their normal secretory function and rescues the mice from early death. As GLUT2 is also expressed in other tissues, these mice were then studied for the presence of other physiological defects due to absence of this transporter. These studies led to the identification of extra-pancreatic, GLUT2-dependent, glucose sensors controlling glucagon secretion and glucose utilization by peripheral tissues, in part through a control of the autonomic nervous system.

Influence of Trypanosoma cruzi strain on the pathogenesis of chronic myocardiopathy in mice

The murine model of chronic Chaga's myocardiopathy was developed in 201 inbred and outbred mice. The experimental groups consisted of 1st: 73 inbred AKR and A/J mice inoculated with one of the following. Trypanosoma cruzi strains: Peruvian (Type I), 12 SF (Type II) or Colombian (Type III); 2nd: 128 outbred Swiss mice, chronically infected either with Type II or Type III strains isolated from human patients from different geographical areas. All T. cruzi strains were previoulsly characterized by their morphobiological behaviour in mice and by isoenzymatic patterns. For the 1st group the inoculum was 5 x 10**4 for the Peruvian strain and 1 x 10**5 for the 12 SF and Colombian strains. In the 2nd group-Swiss mice the inoculum size varied from 2 x 10**4 to 2 x 10**5. The inbred animals were killed at a 3 time-point scale (90, 180 and 240 days) post-infection. The Swiss mice were killed from 180 to 660 days after infection. The evaluation of parasitemia and serology (xeodiagnosis and indirect immunofluorescent test) was performed. The incidence of macroscopic alterations of the heart and cardiac index were evaluated. Histopathological lesions of the myocardium were graded. The influence of T. cruzi strain on the intensity of cardiac lesions was evaluated by the Chi-square test; the incidence of inflammatory lesions and its relationship to the parasite strain was evaluated by the Fisher test. The influence of the duration of infection was evaluated by using the Gamma Coefficient of Kruskal and Goodman and its measure of significance. Slight to severe microscopic alterations occurred in 85% of the chronically infected nice. There were a clear predominance on the incidence and intensity of inflammatory and fibrotic alterations for the mice infected with Type III strains. Statistical analysis has shown significant differences among the infected groups, in the inflammatory and fibrotic lesions. Macroscopic alterations (right cavities dilatation and apex aneurism of left ventricle), differed in incidence according to mice strains; in Swiss and AKR mice, significant differences were seen in mice infected with different T. cruzi strains, but the A/J mice failed to show significant differences correlated with different parasite strains. The duration of infection, from 90 to 240 days, could not be correlated with the degree of lesions in the several groups.

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.

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.

Hypertension pulmonaire associée aux maladies du coeur gauche: quelle définition et quelle prise en charge en 2013 [Pulmonary hypertension in left heart disease: how to define it and how to manage it in 2013?].

Pulmonary hypertension is a frequent complication of left heart disease arising from a wide range of cardiac disorders and is associated with poor prognosis. Its pathophysiology is complex with both passive mechanisms of elevated filling pressures in left cavities and occasionally reactive mechanisms of arterial vasoconstriction and remodelling to interplay. This stage, called <out-of-proportions> pulmonary hypertension, further worsens the heart failure patients' prognosis but is still a matter of debate concerning the criteria to apply for its diagnosis and concerning the best way to manage it. This article gives an overview of the importance and pathophysiology of pulmonary hypertension associated with left heart disease, and discusses the challenges associated with its diagnosis and treatment.