PF-1355, a mechanism-based myeloperoxidase inhibitor, prevents immune complex vasculitis and anti-glomerular basement membrane glomerulonephritis.

Small vessel vasculitis is a life-threatening condition and patients typically present with renal and pulmonary injury. Disease pathogenesis is associated with neutrophil accumulation, activation, and oxidative damage, the latter being driven in large part by myeloperoxidase (MPO), which generates hypochlorous acid among other oxidants. MPO has been associated with vasculitis, disseminated vascular inflammation typically involving pulmonary and renal microvasculature and often resulting in critical consequences. MPO contributes to vascular injury by 1) catabolizing nitric oxide, impairing vasomotor function; 2) causing oxidative damage to lipoproteins and endothelial cells, leading to atherosclerosis; and 3) stimulating formation of neutrophil extracellular traps, resulting in vessel occlusion and thrombosis. Here we report a selective 2-thiouracil mechanism-based MPO inhibitor (PF-1355 [2-(6-(2,5-dimethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide) and demonstrate that MPO is a critical mediator of vasculitis in mouse disease models. A pharmacokinetic/pharmacodynamic response model of PF-1355 exposure in relation with MPO activity was derived from mouse peritonitis. The contribution of MPO activity to vasculitis was then examined in an immune complex model of pulmonary disease. Oral administration of PF-1355 reduced plasma MPO activity, vascular edema, neutrophil recruitment, and elevated circulating cytokines. In a model of anti-glomerular basement membrane disease, formerly known as Goodpasture disease, albuminuria and chronic renal dysfunction were completely suppressed by PF-1355 treatment. This study shows that MPO activity is critical in driving immune complex vasculitis and provides confidence in testing the hypothesis that MPO inhibition will provide benefit in treating human vasculitic diseases.

Comparative Analysis of Virulence Profiles of Serratia marcescens Isolated from Diverse Clinical Origins in Mexican Patients.

Background: Serratia marcescens is an enteric bacterium with increasing incidence in clinical settings, attributed mainly to the opportune expression of diverse virulence determinants plus a wide intrinsic and acquired antibiotic resistance. Methods: The aim of this study was to compare the virulence factor profiles of 185 Serratia marcescens isolates from different clinical origins. In vitro proteolytic and hemolytic activities, biofilm formation, and motility were assessed in each strain. Additionally, the pathogenicity of four hypervirulent strains was analyzed in vivo in Galleria mellonella. Results: We found that bacterial isolates from wound/abscess and respiratory tract specimens exhibited the highest protease activity along with a strong biofilm production, while uropathogenic isolates showed the highest hemolytic activity. Swarming and swimming motilities were similar among all the strains. However, respiratory tract isolates showed the most efficient motility. Two hyperhemolytic and two hyperproteolytic strains were detected; the latter were more efficient killing Galleria mellonella with a 50%-60% larval mortality 48 hours after challenge. Conclusion: A correlation was found between biofilm formation and proteolytic and hemolytic activities in biopsy specimens and bloodstream isolates, respectively. Overall, it becomes critical to evaluate and compare the clinical strains virulence diversity in order to understand the underlying mechanisms that allow the establishment and persistence of opportunistic bacterial infections in the host.

Myeloperoxidase inhibition in mice alters atherosclerotic lesion composition.

Myeloperoxidase (MPO) is a highly abundant protein within the neutrophil that is associated with lipoprotein oxidation, and increased plasma MPO levels are correlated with poor prognosis after myocardial infarct. Thus, MPO inhibitors have been developed for the treatment of heart failure and acute coronary syndrome in humans. 2-(6-(5-Chloro-2-methoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide PF-06282999 is a recently described selective small molecule mechanism-based inactivator of MPO. Here, utilizing PF-06282999, we investigated the role of MPO to regulate atherosclerotic lesion formation and composition in the Ldlr-/- mouse model of atherosclerosis. Though MPO inhibition did not affect lesion area in Ldlr-/- mice fed a Western diet, reduced necrotic core area was observed in aortic root sections after MPO inhibitor treatment. MPO inhibition did not alter macrophage content in and leukocyte homing to atherosclerotic plaques. To assess non-invasive monitoring of plaque inflammation, [18F]-Fluoro-deoxy-glucose (FDG) was administered to Ldlr-/- mice with established atherosclerosis that had been treated with clinically relevant doses of PF-06282999, and reduced FDG signal was observed in animals treated with a dose of PF-06282999 that corresponded with reduced necrotic core area. These data suggest that MPO inhibition does not alter atherosclerotic plaque area or leukocyte homing, but rather alters the inflammatory tone of atherosclerotic lesions; thus, MPO inhibition could have utility to promote atherosclerotic lesion stabilization and prevent atherosclerotic plaque rupture.

Chronic ethanol consumption in rats produces residual increases in anxiety 4 months after withdrawal.

The present study investigated the long-term effects of ethanol consumption in rats. Subjects were maintained on either an ethanol (alcohol) (2.7-6.7%, v/v) or an isocaloric liquid control diet for 26 consecutive days (M=13.7 g/kg/day). Testing for working memory was conducted in a Morris water maze (2 trials/day for 8 days) and commenced after either a short (19 days) or long (120 days) abstinence period. This was followed by assessment of 72 h retention of passive avoidance. Animals were killed either 41 (short abstinence) or 152 days (long abstinence) post-ethanol and their brains stained with cresyl violet. Assessments of dorsal-ventral and medial-lateral cortical vertices were measured in sections derived from eight coronal planes extending +4.20 to -4.16 mm from Bregma. Results indicated that subjects in the ethanol/long abstinence group exhibited increased state anxiety due to their propensity to be thigmotaxtic (i.e., wall-hugging) in the water maze. Unfortunately, such a swim pattern precluded assessment of working memory in our subjects. No evidence of ethanol-induced memory decrements were observed on retention of passive avoidance. There was some evidence that animals in the ethanol/long abstinent group suffered cortical thinning and slight compression of the CA1 layer within the hippocampus, although age might have contributed to the former effect. It was concluded that chronic ethanol consumption increases anxiety even after an extended period of withdrawal and may conspire with age to affect cortical integrity.

2-Aminopyridine-Based Mitogen-Activated Protein Kinase Kinase Kinase Kinase 4 (MAP4K4) Inhibitors: Assessment of Mechanism-Based Safety.

Studies have linked the serine-threonine kinase MAP4K4 to the regulation of a number of biological processes and/or diseases, including diabetes, cancer, inflammation, and angiogenesis. With a majority of the members of our lead series (e.g., 1) suffering from time-dependent inhibition (TDI) of CYP3A4, we sought design avenues that would eliminate this risk. One such approach arose from the observation that carboxylic acid-based intermediates employed in our discovery efforts retained high MAP4K4 inhibitory potency and were devoid of the TDI risk. The medicinal chemistry effort that led to the discovery of this central nervous system-impaired inhibitor together with its preclinical safety profile is described.

Myeloperoxidase Inhibition Improves Ventricular Function and Remodeling After Experimental Myocardial Infarction.

PF-1355 is an oral myeloperoxidase (MPO) inhibitor that successfully decreased elevated MPO activity in mouse myocardial infarction models. Short duration PF-1355 treatment for 7 days decreased the number of inflammatory cells and attenuated left ventricular dilation. Cardiac function and remodeling improved when treatment was increased to 21 days. Better therapeutic effect was further achieved with early compared with delayed treatment initiation (1 h vs. 24 h after infarction). In conclusion, PF-1355 treatment protected a mouse heart from acute and chronic effects of MI, and this study paves the way for future translational studies investigating this class of drugs in cardiovascular diseases.

Metástasis cutánea de carcinoma diferenciado de tiroides. A propósito de un caso / Cutaneous metastasis of differentiated thyroid carcinoma. About a case

El cáncer de tiroides (CT) así como otros tipos de tumores malignos tienen la capacidad de invadir e infiltrar otros tejidos tanto contiguos como tejidos y órganos distantes produciendo lo que se conoce como metástasis (MT). Esta es una característica propia de la mayoría de los tumores cancerígenos, y el mecanismo por el cual lo realizan se produce de distintas maneras dependiendo del tipo histológico del tumor, así como de sus particularidades de diseminación propias sea vía linfática, hematógena, nerviosa o por invasión local. Hablando particularmente del cáncer de tiroides (CT) y siendo más específicos el cáncer papilar (CPT), es la variante histológica maligna que con más prevalencia e incidencia se presenta en esta glándula, además que es el cáncer tiroideo que presenta el mejor pronostico y más alternativas de tratamiento por su patrón de crecimiento lento, y por su baja capacidad de producir metástasis (MT) a distancia, presentado se en menos del 10% de pacientes con este diagnóstico. Pese a esta baja tasa de enfermedad metastásica, esta patología ha presentado un crecimiento notable en su prevalencia llegando a triplicarse en los últimos 30 años, lo que ha originado de la misma manera un aumento en la prevalencia de casos de metástasis (MT) por esta neoplasia. Cuando se presenta enfermedad metastásica del carcinoma papilar de tiroides (CPT) se localiza con mayor frecuencia en tejido ganglionar seguido por tejido pulmonar y óseo. La diseminación de este tipo de cáncer a otros sitios es infrecuente, y a piel es aún más inusual. Por este motivo se presenta el caso de una paciente femenina de 74 años, con antecedentes de diagnóstico de carcinoma diferenciado de tiroides sometida a varias intervenciones quirúrgicas previas, la cual presenta una lesión dérmica a nivel pre esternal, cuyo estudio histopatológico post resección fue compatible con diseminación metastásica cutánea de este tipo de cáncer.

Thyroid cancer (CT) as well as other types of malignant tumors have the ability to invade and infiltrate other tissues both contiguous and distant tissues and organs producing what is known as metastasis (MT). This is a characteristic of the majority of cancerous tumors, and the mechanism by which they are carried out occurs in different ways depending on the histological type of the tumor, as well as its own dissemination characteristics, whether lymphatic, hematogenous, nervous or local invasion. Speaking particularly of thyroid cancer (CT) and being more specific papillary cancer (CPT), it is the malignant histological variant that has the most prevalence and incidence in this gland, in addition it is the thyroid cancer that has the best prognosis and more treatment alternatives due to their slow growth pattern, and due to their low capacity to produce distance metastases (MT) presented in less than 10% of patients with this diagnosis. Despite this low rate of metastatic disease, this pathology has presented a remarkable growth in its prevalence, tripling in the last 30 years, which has also caused an increase in the prevalence of metastatic cases (MT) due to this neoplasia. When there is metastatic disease of papillary thyroid carcinoma (CPT), it is most often located in lymph tissue followed by lung and bone tissue. The spread of this type of cancer to other sites is uncommon, and the skin is even more unusual. For this reason, the case of a 74-year-old female patient is presented, with a history of diagnosis of differentiated thyroid carcinoma undergoing several previous surgical interventions, which presents a dermal lesion at the pre-sternal level, whose post-resection histopathological study was compatible with metastatic skin spread of this type of cancer

Endothelial protein kinase MAP4K4 promotes vascular inflammation and atherosclerosis.

Signalling pathways that control endothelial cell (EC) permeability, leukocyte adhesion and inflammation are pivotal for atherosclerosis initiation and progression. Here we demonstrate that the Sterile-20-like mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4), which has been implicated in inflammation, is abundantly expressed in ECs and in atherosclerotic plaques from mice and humans. On the basis of endothelial-specific MAP4K4 gene silencing and gene ablation experiments in Apoe(-/-) mice, we show that MAP4K4 in ECs markedly promotes Western diet-induced aortic macrophage accumulation and atherosclerotic plaque development. Treatment of Apoe(-/-) and Ldlr(-/-) mice with a selective small-molecule MAP4K4 inhibitor also markedly reduces atherosclerotic lesion area. MAP4K4 silencing in cultured ECs attenuates cell surface adhesion molecule expression while reducing nuclear localization and activity of NFκB, which is critical for promoting EC activation and atherosclerosis. Taken together, these results reveal that MAP4K4 is a key signalling node that promotes immune cell recruitment in atherosclerosis.

Discovery of an in Vivo Tool to Establish Proof-of-Concept for MAP4K4-Based Antidiabetic Treatment.

Recent studies in adipose tissue, pancreas, muscle, and macrophages suggest that MAP4K4, a serine/threonine protein kinase may be a viable target for antidiabetic drugs. As part of the evaluation of MAP4K4 as a novel antidiabetic target, a tool compound, 16 (PF-6260933) and a lead 17 possessing excellent kinome selectivity and suitable properties were delivered to establish proof of concept in vivo. The medicinal chemistry effort that led to the discovery of these lead compounds is described herein together with in vivo pharmacokinetic properties and activity in a model of insulin resistance.

Residual behavioral and neuroanatomical effects of short-term chronic ethanol consumption in rats.

The residual effects of short-term chronic ethanol consumption were investigated in rats maintained on an ethanol liquid diet for 26 consecutive days (mean intake = 16.1 g/kg/day). Animals were assessed for spontaneous motor activity (12 days post-ethanol), spatial working memory (17 days post-ethanol), spatial reference memory (184 days post-ethanol), and retention of passive avoidance (201 days post-ethanol). Measurements of brain weights and cortical thickness vertices within the dorsomedial and ventrolateral cortex of eight coronal planes were determined 260 days post-ethanol. Two-dimensional cell profile densities within six coronal planes and within CA1 region of the hippocampus were also obtained, along with the total volumetric measurement of the hippocampus proper. Results indicated between group differences when subjects were assessed on working memory with ethanol-treated animals exhibiting longer escape latencies in a Morris water maze, an effect partially attributed to the perseverance of ethanol-treated animals in exhibiting thigmotaxicity. No other ethanol-related behavioral impairment was noted. Neuroanatomically, ethanol-treated rats had thinner cortical mantles (6.3% and 6.6% reductions) within the frontoparietal cortex and had lower two-dimensional cell profile densities within the most caudal cortical region studied. Interestingly, control animals with thicker cortical mantles tended to perform better on the working memory task, whereas the opposite was true for ethanol-treated subjects. These data led to the conclusion that chronic ethanol consumption of a relatively short duration produces working memory impairments, albeit mild, that are partially related to an inability to abandon ineffectual behavioral strategies, and also produces neuroanatomical alterations within the cortex.

Conocimiento y valoración de la calidad ambiental de la Reserva Forestal Bosque Yotoco: perspectiva psicoambiental / Knowledge and environmental assessment of forest reserve forest Yotoco quality: environmental psychology perspective

El presente trabajo de investigación se enmarca en el campo de la Psicología Ambiental y tuvo como propósito describir los conocimientos y las valoraciones que las personas expresaron sobre la calidad ambiental de la Reserva Forestal Bosque de Yotoco (Valle del Cauca, Colombia), así como descubrir sugerencias implícitas o explicitas pertinentes para el mejoramiento de la calidad ambiental del ecosistema en mención. Los participantes fueron algunos de los visitantes a la reserva desde el año 1994 hasta el 2011. Este estudio utilizó una estrategia metodológica cualitativa. Los datos analizados se obtuvieron a través del muestreo al azar de los libros de registro de visitantes, en el cual éstos escriben los comentarios sobre el carácter de su experiencia inmediatamente culminan el recorrido de la misma. Los resultados son ampliamente favorables hacia el reconocimiento de la calidad ambiental de la Reserva en sus aspectos físicos, bióticos y especialmente los psicosociales. Se destacan algunos comentarios relacionados con la apreciación del paisaje, la importancia como fuente originaria y regula-dora de agua, el ser uno de los pocos "sobrevivientes" de bosque en la vertiente oriental de la cordillera occidental colombiana, la buena atención de los funcionarios y su posibilidad para la educación ambiental pertinente.

The present research is framed in the field of environmental psychology and was aimed to describe the knowledge and assessments that people expressed about the environmental quality of the forest Forest Reserve Yotoco (Colombia), and discover relevant implicit or explicit suggestions to improve the environmental quality of the ecosystem in question. Participants were among the visitors to the Reserve during the years 1994 through 2011. This study used a qualitative methodological strategy. The data analyzed were obtained through random sampling of the record books of visitors where they write comments on the nature of their experience immediately culminate the course of it. The results are very favorable towards the recognition of the environmental quality of the Reserve in its physical, biotic and especially psychosocial aspects. The study highlights some reviews related to the appreciation of the landscape, the importance as originating and regulating water supply, one of the few to be "survivors" of forest on the eastern slope of the Colombian Western Cordillera, good care provided by the staff and the possibility for the relevant environmental education.

Intracellular ascorbic acid inhibits transport of glucose by neurons, but not by astrocytes.

It has been demonstrated that glutamatergic activity induces ascorbic acid (AA) depletion in astrocytes. Additionally, different data indicate that AA may inhibit glucose accumulation in primary cultures of rat hippocampal neurons. Thus, our hypothesis postulates that AA released from the astrocytes during glutamatergic synaptic activity may inhibit glucose uptake by neurons. We observed that cultured neurons express the sodium-vitamin C cotransporter 2 and the facilitative glucose transporters (GLUT) 1 and 3, however, in hippocampal brain slices GLUT3 was the main transporter detected. Functional activity of GLUTs was confirmed by means of kinetic analysis using 2-deoxy-d-glucose. Therefore, we showed that AA, once accumulated inside the cell, inhibits glucose transport in both cortical and hippocampal neurons in culture. Additionally, we showed that astrocytes are not affected by AA. Using hippocampal slices, we observed that upon blockade of monocarboxylate utilization by alpha-cyano-4-hydroxycinnamate and after glucose deprivation, glucose could rescue neuronal response to electrical stimulation only if AA uptake is prevented. Finally, using a transwell system of separated neuronal and astrocytic cultures, we observed that glutamate can reduce glucose transport in neurons only in presence of AA-loaded astrocytes, suggesting the essential role of astrocyte-released AA in this effect.

Sodium vitamin C cotransporter SVCT2 is expressed in hypothalamic glial cells.

Kinetic analysis of vitamin C uptake demonstrated that different specialized cells take up ascorbic acid through sodium-vitamin C cotransporters. Recently, two different isoforms of sodium-vitamin C cotransporters (SVCT1/SLC23A1 and SVCT2/SLC23A2) have been cloned. SVCT2 was detected mainly in choroidal plexus cells and neurons; however, there is no evidence of SVCT2 expression in glial and endothelial cells of the brain. Certain brain locations, including the hippocampus and hypothalamus, consistently show higher ascorbic acid values compared with other structures within the central nervous system. However, molecular and kinetic analysis addressing the expression of SVCT transporters in cells isolated from these specific areas of the brain had not been done. The hypothalamic glial cells, or tanycytes, are specialized ependymal cells that bridge the cerebrospinal fluid with different neurons of the region. Our hypothesis postulates that SVCT2 is expressed selectively in tanycytes, where it is involved in the uptake of the reduced form of vitamin C (ascorbic acid), thereby concentrating this vitamin in the hypothalamic area. In situ hybridization and optic and ultrastructural immunocytochemistry showed that the transporter SVCT2 is highly expressed in the apical membranes of mouse hypothalamic tanycytes. A newly developed primary culture of mouse hypothalamic tanycytes was used to confirm the expression and function of the SVCT2 isoform in these cells. The results demonstrate that tanycytes express a high-affinity transporter for vitamin C. Thus, the vitamin C uptake mechanisms present in the hypothalamic glial cells may perform a neuroprotective role concentrating vitamin C in this specific area of the brain.

Neumonía y falla respiratoria por influenza AHINl en la unidad de cuidados intensivos del Hospital Metropolitano de Quito / Pneumonia and influenza AHINl respiratory failure in the intensive care unit of Hospital Metropolitano Quito

A finales de marzo de 2009, se produjo el brote de una enfermedad respiratoria en el poblado de La Gloria en México. Un mes más tarde esta infección respiratoria resultó ser la llamada "gripe porcina" producida por un agente etiológico nuevo, el virus influenza AHINI. Esta enfermedad rápidamente se diseminó hacia los distintos paises del mundo facilitados por el desarrollo de los medios de transporte, principalmente el aéreo. Es así cómo en el mes de mayo el primer caso de gripe porcina arribó al Ecuador en la provincia de Guayas, en pacientes que llegaron desde los Estados Unidos; desde este punto se diseminó rapidamente y cerca de varios miles de personas la han contraído en nuestro país. Sus características clínicas son las de una enfermedad respiratoria caracterizada por cursar con la sintomatología de un proceso inflamatorio leve hasta una franca falla respiratoria grave, asociada a daño alveolar. En este artículo se describen las características clínicas y epidemiológicas de las personas hospitalizadas por neumonía rápidamente progresiva en el Servicio de Cuidados Intensivos del Hospítal Metropolitano de Quito.