Insights on the genetic features of endometrial pathogenic Escherichia coli strains from pyometra in companion animals: Improving the knowledge about pathogenesis.

Endometrial pathogenic E. coli (EnPEC) isolates are involved in endometrial infections in animals and humans. Besides the high aggressiveness of the endometrial infections, the EnPEC virulence profile and pathogenesis are still little known. In this study, we have sequenced and analyzed an EnPEC strain from canine pyometra (E. coli_LBV005/17), following a molecular characterization of the virulence profile and phylogenetic evolution of an EnPEC collection from canines and felines (45 strains). Most of the strains belonged to phylo-group B2, and display a high virulence profile. In particular we highlight the classification of the E. coli_LBV005/17 as sequence type 131 (ST131), in addition to other five strains, as observed by gyrB phylogenetic analysis. Also, the phylogenetic position of EnPEC strains from pyometra in companion animals suggests that their origins are from both extraintestinal and commensal E. coli strains. Accordingly to Principal Coordinates Analysis (PCoA) and phylogenetic analysis we can propose that EnPEC strains have neither the same genetic profile, nor a unique common ancestral. In summary, the present work characterize an EnPEC genome from bitch pyometra and the genetic profile of 45 EnPEC strains from companion animals pyometra, being the commonest virulence pattern: fimA, papC, hlyA, hlyE, cnf1, entB, iroN, irp1, bssS, bssR, and hmsP. These data improving the background knowledge of this E. coli pathotype related to pyometra in companion animals and may support new methods to prevent the disease evolution.

RANKL Triggers Treg-Mediated Immunoregulation in Inflammatory Osteolysis.

The chronic inflammatory immune response triggered by the infection of the tooth root canal system results in the local upregulation of RANKL, resulting in periapical bone loss. While RANKL has a well-characterized role in the control of bone homeostasis/pathology, it can play important roles in the regulation of the immune system, although its possible immunoregulatory role in infectious inflammatory osteolytic conditions remains largely unknown. Here, we used a mouse model of infectious inflammatory periapical lesions subjected to continuous or transitory anti-RANKL inhibition, followed by the analysis of lesion outcome and multiple host response parameters. Anti-RANKL administration resulted in arrest of bone loss but interfered in the natural immunoregulation of the lesions observed in the untreated group. RANKL inhibition resulted in an unremitting proinflammatory response, persistent high proinflammatory and effector CD4 response, decreased regulatory T-cell (Treg) migration, and lower levels of Treg-related cytokines IL-10 and TGFb. Anti-RANKL blockade impaired the immunoregulatory process only in early disease stages, while the late administration of anti-RANKL did not interfere with the stablished immunoregulation. The impaired immunoregulation due to RANKL inhibition is characterized by increased delayed-type hypersensitivity in vivo and T-cell proliferation in vitro to the infecting bacteria, which mimic the effects of Treg inhibition, reinforcing a possible influence of RANKL on Treg-mediated suppressive response. The adoptive transfer of CD4+FOXp3+ Tregs to mice receiving anti-RANKL therapy restored the immunoregulatory capacity, attenuating the inflammatory response in the lesions, reestablishing normal T-cell response in vivo and in vitro, and preventing lesion relapse upon anti-RANKL therapy cessation. Therefore, while RANKL inhibition efficiently limited the periapical bone loss, it promoted an unremitting host inflammatory response by interfering with Treg activity, suggesting that this classic osteoclastogenic mediator plays a role in immunoregulation.

Endolymphatic sac drainage for the treatment of Ménière's disease.

OBJECTIVE: To describe the results obtained with endolymphatic sac drainage in patients with Ménière's disease. METHOD: A retrospective case review study was conducted of 95 Ménière's disease patients who underwent endolymphatic sac drainage in a tertiary care referral centre, after failing a long course of medical management. The main outcome measures were vertigo control and hearing preservation. RESULTS: In patients with unilateral disease, vertigo control was obtained in 94.3 per cent of patients. A significant improvement in cochlear function was seen in 14 per cent of patients, and hearing was preserved or improved in 88 per cent. For the bilateral group, vertigo control was obtained in 85.7 per cent of patients and cochlear function improved in 28 per cent. Hearing preservation was attained in 71 per cent of these patients. CONCLUSION: Endolymphatic sac drainage is a good surgical option for patients with incapacitating endolymphatic hydrops, providing a high percentage of vertigo control and hearing preservation.

Glioblastoma cells inhibit astrocytic p53-expression favoring cancer malignancy.

The tumor microenvironment has a dynamic and usually cancer-promoting function during all tumorigenic steps. Glioblastoma (GBM) is a fatal tumor of the central nervous system, in which a substantial number of non-tumoral infiltrated cells can be found. Astrocytes neighboring these tumor cells have a particular reactive phenotype and can enhance GBM malignancy by inducing aberrant cell proliferation and invasion. The tumor suppressor p53 has a potential non-cell autonomous function by modulating the expression of secreted proteins that influence neighbor cells. In this work, we investigated the role of p53 on the crosstalk between GBM cells and astrocytes. We show that extracellular matrix (ECM) from p53(+/-) astrocytes is richer in laminin and fibronectin, compared with ECM from p53(+/+) astrocytes. In addition, ECM from p53(+/-) astrocytes increases the survival and the expression of mesenchymal markers in GBM cells, which suggests haploinsufficient phenotype of the p53(+/-) microenvironment. Importantly, conditioned medium from GBM cells blocks the expression of p53 in p53(+/+) astrocytes, even when DNA was damaged. These results suggest that GBM cells create a dysfunctional microenvironment based on the impairment of p53 expression that in turns exacerbates tumor endurance.

Activated microglia mediate synapse loss and short-term memory deficits in a mouse model of transthyretin-related oculoleptomeningeal amyloidosis.

Oculoleptomeningeal amyloidosis (OA) is a fatal and untreatable hereditary disease characterized by the accumulation of transthyretin (TTR) amyloid within the central nervous system. The mechanisms underlying the pathogenesis of OA, and in particular how amyloid triggers neuronal damage, are still unknown. Here, we show that amyloid fibrils formed by a mutant form of TTR, A25T, activate microglia, leading to the secretion of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and nitric oxide. Further, we found that A25T amyloid fibrils induce the activation of Akt, culminating in the translocation of NFκB to the nucleus of microglia. While A25T fibrils were not directly toxic to neurons, the exposure of neuronal cultures to media conditioned by fibril-activated microglia caused synapse loss that culminated in extensive neuronal death via apoptosis. Finally, intracerebroventricular (i.c.v.) injection of A25T fibrils caused microgliosis, increased brain TNF-α and IL-6 levels and cognitive deficits in mice, which could be prevented by minocycline treatment. These results indicate that A25T fibrils act as pro-inflammatory agents in OA, activating microglia and causing neuronal damage.

PLP1 duplication at the breakpoint regions of an apparently balanced t(X;22) translocation causes Pelizaeus-Merzbacher disease in a girl.

PLP1 (proteolipid protein1 gene) mutations cause Pelizaeus-Merzbacher disease (PMD), characterized by hypomyelination of the central nervous system, and affecting almost exclusively males. We report on a girl with classical PMD who carries an apparently balanced translocation t(X;22)(q22;q13). By applying array-based comparative genomic hybridization (a-CGH), we detected duplications at 22q13 and Xq22, encompassing 487-546 kb and 543-611 kb, respectively. The additional copies were mapped by fluorescent in situ hybridization to the breakpoint regions, on the derivative X chromosome (22q13 duplicated segment) and on the derivative 22 chromosome (Xq22 duplicated segment). One of the 14 duplicated X-chromosome genes was PLP1.The normal X chromosome was the inactive one in the majority of peripheral blood leukocytes, a pattern of inactivation that makes cells functionally balanced for the translocated segments. However, a copy of the PLP1 gene on the derivative chromosome 22, in addition to those on the X and der(X) chromosomes, resulted in two active copies of the gene, irrespective of the X-inactivation pattern, thus causing PMD. This t(X;22) is the first constitutional human apparently balanced translocation with duplications from both involved chromosomes detected at the breakpoint regions.

Microglial stress inducible protein 1 promotes proliferation and migration in human glioblastoma cells.

Microglial activation is a key event in the progression and infiltration of tumors. We have previously demonstrated that the co-chaperone stress inducible protein 1 (STI1), a cellular prion protein (PrP(C)) ligand, promotes glioblastoma (GBM) proliferation. In the present study, we examined the influence of microglial STI1 in the growth and invasion of the human glioblastoma cell line GBM95. We demonstrated that soluble factors secreted by microglia into the culture medium (microglia conditioned medium; MG CM) caused a two-fold increase in the proliferation of GBM95 cells. This effect was reversed when STI1 was removed from the MG CM. In this context, we have shown that microglial cells synthesize and secrete STI1. Interestingly, no difference was observed in proliferation rates when GBM cells were maintained in MG CM or MG CM containing an anti-PrP(C) neutralizing antibody. Moreover, rec STI1 and rec STI1(Δ230-245), which lack the PrP(C) binding site, both promoted similar levels of GBM95 proliferation. In the migration assays, MG CM favored the migration of GBM95 cells, but migration failed when STI1 was removed from the MG CM. We detected metalloproteinase 9 (MMP-9) activity in the MG CM, and when cultured microglia were treated with an anti-STI1 antibody, MMP-9 activity decreased. Our results suggest that STI1 is secreted by microglia and favors tumor growth and invasion through the participation of MMP-9 in a PrP(C)-independent manner.

Marine habitats map of "Isla del Caño", Costa Rica, comparing Quickbird and Hymap images classification results.

Isla del Caño is a marine protected area on the south Pacific coast of Costa Rica, surrounded by coral reefs and coral communities. The ecology of these coral reefs has been studied for over 20 years, but they have not been mapped. Maps are considered a great research, planning, management and monitoring tool. Medium to high resolution images (HyMap 2005 and Quickbird 2007 respectively) were processed and classified in order to test and compare their accuracy in producing a marine habitat map. Manta tow surveys were performed in the field for identification of 7 marine habitats 1. Coral community; 2. Coral reef; 3. Bed rock; 4. Sand; 5. Sand with boulders; 6. Sand with rodolyths; and 7. Deep water. The overall accuracy was slightly higher using Quickbird (87%) than using Hymap (60%), however the difference was not significant. The map produced using Quickbird was selected to represent the marine habitats of Isla del Caño. This map will help to analyze the adequate size and zoning of the marine protected area.

Marine habitats map of "Isla del Caño", Costa Rica, comparing Quickbird and Hymap images classification results

Isla del Caño is a marine protected area on the south Pacific coast of Costa Rica, surrounded by coral reefs and coral communities. The ecology of these coral reefs has been studied for over 20 years, but they have not been mapped. Maps are considered a great research, planning, management and monitoring tool. Medium to high resolution images (HyMap 2005 and Quickbird 2007 respectively) were processed and classified in order to test and compare their accuracy in producing a marine habitat map. Manta tow surveys were performed in the field for identification of 7 marine habitats 1. Coral community; 2. Coral reef; 3. Bed rock; 4. Sand; 5. Sand with boulders; 6. Sand with rodolyths; and 7. Deep water. The overall accuracy was slightly higher using Quickbird (87%) than using Hymap (60%), however the difference was not significant. The map produced using Quickbird was selected to represent the marine habitats of Isla del Caño. This map will help to analyze the adequate size and zoning of the marine protected area. Rev. Biol. Trop. 58 (1): 373-381. Epub 2010 March 01.

Isla del Caño es un área marina protegida en la costa del Pacífico de Costa Rica y está rodeada de arrecifes coralinos. La ecología de estos arrecifes coralinos ha sido estudiada a lo largo de 20 años pero todavía no habían sido mapeados. Los mapas son considerados una buena herramienta de investigación, planificación, manejo y monitoreo. Imágenes de mediana y alta resolución (Hymap 2005 y Quickbird 2007 respectivamente) fueron procesadas y clasificadas con el fin de evaluar y comparar su desempeño en la elaboración de un mapa de hábitats marinos. En el campo se realizaron sondeos tipo Manta para la identificación de siete hábitats marinos: 1. Comunidad coralina; 2. Arrecife coralino; 3. Roca; 4. Arena; 5. Arena con cantos; 6. Arena con rodolitos; y 7. Agua profunda. La exactitud de la clasificación resultó un poco mayor usando Quickbird (87%) que Hymap (60%), sin embargo la diferencia no era significativa. Se seleccionó el mapa resultante de la clasificación con Quickbird para representar los hábitats marinos de Isla del Caño. Este mapa puede ayudar a analizar el tamaño adecuado y la zonificación del área marina protegida.

Non-colonial coral macro-borers as indicators of coral reef status in the South Pacific of Costa Rica.

Coral reef status was surveyed in three south Pacific coral reefs of Costa Rica, one in Caño Island and two in Golfo Dulce, and the density, richness and distribution of non-colonial macro borers (> 1 mm) was determined in dead and live coral fragments from these reefs. Based upon traditional indicators of degradation such as high particulate suspended matter and low live coral cover, the reefs at Caño Island are in better condition than those at Golfo Dulce. Reef degradation in Golfo Dulce is mainly due to high loads of terrestrial sediments as a consequence of watersheds deforestation. In this study, 36 coral boring species are reported for the eastern Pacific. At the family level, there is high endemism (10%) and greater affinity with the Indo-Pacific (34%), as compared with the eastern Atlantic and Mediterranean (29%) and western Atlantic and Caribbean (27%). The dominant non-colonial macro boring families at the study reefs are mytilid bivalves, eunicid polychaetes and aspidosiphonid sipunculans, with the bivalves considered the main internal bioeroders due to their greater body size and abundances. The level of mortality of the coral colonies and the general level of reef degradation influenced the composition of non-colonial macro-borers. Diversity and total macro-borer density, especially aspidosiphonid density, is higher in corals with greates dead than live cover. In the healthiest coral colonies (less than 50% of partial mortality), mytilids domination, macro-borer diversity and total density, is higher in Golfo Dulce, where reefs are more degraded. In the most affected coral colonies (more than 50% dead), macro-borers total density, especially aspidosiphonids density, is higher, of the healthiest reef of this study, Platanillo. Bivalve relative abundance increases and sipunculan relative abundance decreases with increasing site degradation. In conclusion bioeroder variables can also be used as reef health indicators.

[The mangrove and others vegetation associations in de Gandoca lagoon, Limón, Costa Rica]. / El manglar y otras asociaciones vegetales de la laguna de Gandoca, Limón, Costa Rica.

Six plant associations were identified at Gandoca Lagoon by photointerpretation and field verification: a) mangroves, b) palm trees swamp, and palm trees with Acrostichum aureum and A. danaefolium, c) mixed palm trees, d) very humid tropical rain forest, and e) tropical beach vegetation. The mangroves cover 12.5 ha surrounding the lagoon and extend 2 km up the Gandoca River. Rhizophora mangle (red mangrove) was the dominant species, with Avicennia germinans (black mangrove), Laguncularia racemosa (white mangrove) and Conocarpus erectus (buttonwood) also present. Moving inland the mangroves grade into a tropical rain forest. Gandoca, the largest and best preserved mangrove of Caribbean Costa Rica, tripled its area from 1976 to 2000. Possible causes include sedimentation and the Limón earthquake, which may have subside the lagoon area.

[Race and left ventricle hypertrophy without hypertension and pulmonary heart disease]. / Raça e hipertrofia do ventrículo esquerdo na ausência de hipertensão e de doenças cardiopulmonares.

PURPOSE: To analyze the association between black people and left ventricular hypertrophy (LVH) in the absence of hypertension and/or other cardiopulmonar disease. METHODS: Data were collected from necropsies carried out in the Anatomopathologic Service (APS). Hospital Edgard Santos from 1970 to 1986, Salvador. It were included only subjects at ages > or = 20 years and free of hypertension and any cardiopulmonar disease. A LV wall > 1.6cm was considered as LVH (standardized criteria of the APS). Controls variables were age, sex, and absence of the mentioned diseases. It was used a case-control epidemiological study design and the association measured by "odds ratio" (OR) for no matched case-control study. RESULTS: From the 208 subjects studied, 48 (23.1%) had LVH. There was no difference in the frequency of right ventricular hypertrophy between cases and controls (p > 0.05). The mean of heart weight was higher for LVH cases (p < 0.001), but there was no evidence of association between blacks and LVH (OR = 1.05, p > 0.05 and confidence interval at 95% = 0.8, 1.31. The highest odds possible for the association in this study (assuming that all 3 LVH losses were black subjects) would be 1.5, also no statistically significant. CONCLUSION: In the absence of hypertension and other cardiopulmonar diseases, LHV is common in necropsies in Salvador, Brazil, with similar frequencies in blacks, whites and mullatos and seems not be a risk factor for hypertension in black people.