Matrix metalloproteinase-10 is upregulated by thrombin in endothelial cells and increased in patients with enhanced thrombin generation

OBJECTIVE: Thrombin is a multifunctional serine protease that promotes vascular proinflammatory responses whose effect on endothelial MMP-10 expression has not previously been evaluated. METHODS AND RESULTS: Thrombin induced endothelial MMP-10 mRNA and protein levels, through a protease-activated receptor-1 (PAR-1)-dependent mechanism, in a dose- and time-dependent manner. This effect was mimicked by a PAR-1 agonist peptide (TRAP-1) and antagonized by an anti-PAR-1 blocking antibody. MMP-10 induction was dependent on extracellular regulated kinase1/2 (ERK1/2) and c-jun N-terminal kinase (JNK) pathways. By serial deletion analysis, site-directed mutagenesis and electrophoretic mobility shift assay an AP-1 site in the proximal region of MMP-10 promoter was found to be critical for thrombin-induced MMP-10 transcriptional activity. Thrombin and TRAP-1 upregulated MMP-10 in murine endothelial cells in culture and in vivo in mouse aorta. This effect of thrombin was not observed in PAR-1-deficient mice. Interestingly, circulating MMP-10 levels (P<0.01) were augmented in patients with endothelial activation associated with high (disseminated intravascular coagulation) and moderate (previous acute myocardial infarction) systemic thrombin generation. CONCLUSIONS: Thrombin induces MMP-10 through a PAR-1-dependent mechanism mediated by ERK1/2, JNK, and AP-1 activation. Endothelial MMP-10 upregulation could be regarded as a new proinflammatory effect of thrombin whose pathological consequences in thrombin-related disorders and plaque stability deserve further investigation

New records of Peters squirrel Sciurus oculatus (Peters 1863) for Michoacán and first molecular genotyping of the species

Capturamos tres ejemplares de la ardilla de Peters (Sciurus oculatus), especie listada en protección especial y endémica del centro de México. Las capturas ocurrieron en fragmentos de bosque de encino del municipio de Huandacareo, Michoacán. Con el registro de esta población se confirma la presencia actual de la especie en el estado, ya que su último registro es de 1986 de un ejemplar colectado en Contepec. La localidad de Huandacareo extiende la distribución conocida de la especie hacia el oeste en 48 km aproximadamente con respecto a su registro más reciente. Al revisar los pocos registros de la especie para el estado, se observó que el registro más occidental de la especie, el de Ziracuaretiro (cerca de Uruapan), no se incluyó en la delimitación de su distribución, por lo que el mapa de distribución conocida debe actualizarse incluyendo todas las localidades, lo que modificaría de forma importante su distribución. Revisando la base de datos de GeneBank, notamos la inexistencia de secuencias de ADN mitocondrial de S. oculatus. En este estudio presentamos las dos primeras secuencias del gen de citocromo b (cytb) provenientes de dos individuos de S. oculatus. Las secuencias fueron comparadas en análisis filogenético con ocho especies del género Sciurus disponibles en GeneBank. En el análisis de las secuencias y árbol de UPGMA que incluyó especies de Norteamérica y de Sudamerica, S. oculatus se agrupó con S. aestuans y S. stramineus de Sudamérica, siendo S. oculatus la especie basal. Es importante que en el futuro se investigue con mayor detalle la relación de la especie en el género Sciurus. Desde la perspectiva de  conservación debe evaluarse su distribución actual en el país. Actualmente se desconoce cuántas poblaciones existen en áreas protegidas de México. La población registrada en Huandacareo parece estar aislada y su hábitat rodeado de agricultura.We captured three individuals of Peter’s squirrel (Sciurus oculatus), species listed under special protection and endemic for central Mexico. The captures occurred on oak forest fragments from Huandacareo municipality, Michoacán. With the record of this population, we confirm the species actual presence for the state, because the last record corresponded to a specimen collected in Contepec in 1986. This locality extends the species known distribution to the west approximately in 48 km from its last known record. When we reviewed the species historical records for the state, we notice that the western-most record of the species, a record from Ziracuaretiro (near to Uruapan), was not included to outline the species global distribution, therefore we consider that the present distributional map should be updated including all the localities, which will modify its distribution considerably. After surveying the GeneBank international database, we noticed that mitocondrial DNA sequences from S. oculatus were inexistent. In this work we presented the first two cytochrome b gene (cytb) sequences from two S. oculatus individuals. The sequences were compared with eight species of the Sciurus genus available in GeneBank. In the sequences analysis and UPGMA tree, which included Sciurus species from NorthAmerica and South America, S. oculatus was clustered with S. aestuans and S. stramineus from South America, resulting S. oculatus the basal species. From the conservation perspective, the species actual presencein the historical localities should be verified. No protected areas in the country are known toinclude Sciurus oculatus populations, and the Huandacareo recorded population seems to be relatively isolated, and its habitat surrounded by agriculture

Matrix metalloproteinase-9, -10, and tissue inhibitor of matrix metalloproteinases-1 blood levels as biomarkers of severity and mortality in sepsis

INTRODUCTION: Matrix metalloproteinases (MMPs) play a role in infectious diseases through extracellular matrix (ECM) degradation, which favors the migration of immune cells from the bloodstream to sites of inflammation. Although higher levels of MMP-9 and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) have been found in small series of patients with sepsis, MMP-10 levels have not been studied in this setting. The objective of this study was to determine the predictive value of MMP-9, MMP-10, and TIMP-1 on clinical severity and mortality in a large series of patients with severe sepsis. METHODS: This was a multicenter, observational, and prospective study carried out in six Spanish Intensive Care Units. We included 192 (125 surviving and 67 nonsurviving) patients with severe sepsis and 50 age- and sex-matched healthy controls in the study. Serum levels of MMP-9, MMP-10, TIMP-1, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-10 were measured in patients with severe sepsis at the time of diagnosis and in healthy controls. RESULTS: Sepsis patients had higher levels of MMP-10 and TIMP-1, higher MMP-10/TIMP-1 ratios, and lower MMP-9/TIMP-1 ratios than did healthy controls (P < 0.001). An association was found between MMP-9, MMP-10, TIMP-1, and MMP-9/TIMP-1 ratios and parameters of sepsis severity, assessed by the SOFA score, the APACHE-II score, lactic acid, platelet count, and markers of coagulopathy. Nonsurviving sepsis patients had lower levels of MMP-9 (P = 0.037), higher levels of TIMP-1 (P < 0.001), lower MMP-9/TIMP-1 ratio (P = 0.003), higher levels of IL-10 (P < 0.001), and lower TNF-alpha/IL-10 ratio than did surviving patients. An association was found between MMP-9, MMP-10, and TIMP-1 levels, and TNF-alpha and IL-10 levels. The risk of death in sepsis patients with TIMP-1 values greater than 531 ng/ml was 80% higher than that in patients with lower values (RR = 1.80; 95% CI = 1.13 to 2.87;P = 0.01; sensitivity = 0.73; specificity = 0.45). CONCLUSIONS: The novel findings of our study on patients with severe sepsis (to our knowledge, the largest series reporting data about MMP levels in sepsis) are that reduced MMP-9/TIMP-1 ratios and increased MMP-10 levels may be of great pathophysiologic significance in terms of severity and mortality, and that TIMP-1 levels may represent a biomarker to predict the clinical outcome of patients with sepsis

Association between serum soluble CD40 ligand levels and mortality in patients with severe sepsis

INTRODUCTION: CD40 Ligand (CD40L) and its soluble counterpart (sCD40L) are proteins that exhibit prothrombotic and proinflammatory properties on binding to their cell surface receptor CD40. The results of small clinical studies suggest that sCD40L levels could play a role in sepsis; however, there are no data on the association between sCD40L levels and mortality of septic patients. Thus, the aim of this study was to determine whether circulating sCD40L levels could be a marker of adverse outcome in a large cohort of patients with severe sepsis. METHODS: This was a multicenter, observational and prospective study carried out in six Spanish intensive care units. Serum levels of sCD40L, tumour necrosis factor-alpha and interleukin-10, and plasma levels of tissue factor were measured in 186 patients with severe sepsis at the time of diagnosis. Serum sCD40L was also measured in 50 age- and sex-matched controls. Survival at 30 days was used as the endpoint. RESULTS: Circulating sCD40L levels were significantly higher in septic patients than in controls (P = 0.01), and in non-survivors (n = 62) compared to survivors (n = 124) (P = 0.04). However, the levels of CD40L were not different regarding sepsis severity. Logistic regression analysis showed that sCD40L levels >3.5 ng/mL were associated with higher mortality at 30 days (odds ratio = 2.89; 95% confidence interval = 1.37 to 6.07; P = 0.005). The area under the curve of sCD40L levels >3.5 ng/mL as predictor of mortality at 30 days was 0.58 (95% CI = 0.51 to 0.65; P = 0.03). CONCLUSIONS: In conclusion, circulating sCD40L levels are increased in septic patients and are independently associated with mortality in these patients; thus, its modulation could represent an attractive therapeutic target

Association of sepsis-related mortality with early increase of TIMP-1/MMP-9 ratio

OBJECTIVE: Higher circulating levels of tissue inhibitor of matrix metalloproteinases (TIMP)-1 at the time of severe sepsis diagnosis have been reported in nonsurviving than in surviving patients. However, the following questions remain unanswered: 1) Does TIMP-1/MMP-9 ratio differ throughout the first week of intensive care between surviving and non-surviving patients? 2) Is there an association between TIMP-1/MMP-9 ratio and sepsis severity and mortality during such period? 3) Could TIMP-1/MMP-9 ratio during the first week be used as an early biomarker of sepsis outcome? 4) Is there an association between TIMP-1/MMP-9 ratio and coagulation state and circulating cytokine levels during the first week of intensive care in these patients? The present study sought to answer these questions. METHODS: Multicenter, observational and prospective study carried out in six Spanish Intensive Care Units (ICUs) of 295 patients with severe sepsis. Were measured circulating levels of TIMP-1, MMP-9, tumour necrosis factor (TNF)-alpha, interleukin (IL)-10 and plasminogen activator inhibitor (PAI)-1 at day 1, 4 and 8. End-point was 30-day mortality. RESULTS: We found higher TIMP-1/MMP-9 ratio during the first week in non-surviving (n = 98) than in surviving patients (n = 197) (p<0.01). Logistic regression analyses showed that TIMP-1/MMP-9 ratio at days 1, 4 and 8 was associated with mortality. Receiver operating characteristic (ROC) curves showed that TIMP-1/MMP-9 ratio at days 1, 4 and 8 could predict mortality. There was an association between TIMP-1/MMP-9 ratio and TNF-alpha, IL-10, PAI-1 and lactic acid levels, SOFA score and platelet count at days 1, 4 and 8. CONCLUSIONS: The novel findings of our study were that non-surviving septic patients showed persistently higher TIMP-1/MMP-9 ratio than survivors ones during the first week, which was associated with severity, coagulation state, circulating cytokine levels and mortality; thus representing a new biomarker of sepsis outcome

The 372 T/C genetic polymorphism of TIMP-1 is associated with serum levels of TIMP-1 and survival in patients with severe sepsis

Introduction: Previous studies have found higher circulating levels of tissue inhibitor of matrix metalloproteinase (TIMP)-1 in nonsurviving septic patients than in surviving septic patients, and an association between the 372 T/C genetic polymorphism of TIMP-1 and the risk of developing certain diseases. However, the relationship between genetic polymorphisms of TIMP-1, circulating TIMP-1 levels and survival in patients with severe sepsis has not been examined, and this was the objective of the study. Methods: This multicentre, prospective, observational study was carried out in six Spanish ICUs. We determined the 372 T/C genetic polymorphism of TIMP-1 (rs4898), serum levels of TIMP-1, matrix metalloproteinase (MMP)-9, MMP-10, TNFa, IL-10 and plasma plasminogen activator inhibitor-1 (PAI-1). Survival at 30 days from ICU admission was the endpoint assessed. The association between continuous variables was carried out using Spearman’s rank correlation coefficient or Spearman’s rho coefficient. Multivariate logistic regression analysis was applied to determine the association between the 372 T/C genetic polymorphism and survival 30 days from ICU admission. Results: Of 275 patients with severe sepsis, 80 had genotype CC, 55 had genotype CT and 140 had genotype TT of the 372 T/C genetic polymorphism of TIMP-1. Patients with the T allele showed higher serum levels of TIMP-1 than patients without the T allele (P = 0.004). Multiple logistic regression analysis showed that the T allele was associated with higher mortality at 30 days (odds ratio = 2.08; 95% confidence interval = 1.06 to 4.09; P = 0.03). Survival analysis showed that patients with the T allele presented lower 30-day survival than patients without the T allele (c2 = 5.77; P = 0.016). We found an association between TIMP-1 levels and levels of MMP-9 (r = -0.19; P = 0.002), MMP-10 (r = 0.55; P <0.001), TNFa (r = 0.56; P <0.001), IL-10 (r = 0.48; P <0.001) and PAI-1 (r = 0.49; P <0.001). Conclusion: The novel findings of our study are that septic patients with the T allele in the 372 T/C genetic polymorphism of TIMP-1 showed higher serum TIMP-1 levels and lower survival rate. The determination of the 372 T/C genetic polymorphism of TIMP-1 thus has prognostic implications and could help in the selection of patients who may benefit from modulation of the MMP/TIMP balance

Parathyroid hemangioma

Background: Hemangiomas are benign neoplasms of capillary proliferation that arise from a developmental anomaly where angioblastic mesenchyme fails to form canals. Most hemangiomas arise in the head and neck region, either superficially in the skin or deeper within endocrine organs such as the parotid gland. Parathyroid hemangiomas, however, are extremely rare, with only five cases previously reported in the literature. Case presentation: Herein, we present a case of a 68-year-old man with a hemangioma almost completely replacing the right upper parathyroid gland, grossly measuring 1.3 × 1.3 × 1.2 cm and weighing 700 mg, associated with primary hyperparathyroidism. Conclusions: Parathyroid gland enlargement due to vascular neoplasms such as hemangiomas can mimic, both clinically and radiographically, hyperplasias and/or adenomas. Surgeons need to be aware of the presence of this entity and should consider it in the differential diagnosis of hyperparathyroidism or parathyroid gland enlargement

<i>PIK3R1</i>, <i>HRAS</i> and <i>AR</i> Gene Alterations Associated with Sclerosing Polycystic Adenoma of the Parotid Gland

Sclerosing polycystic adenoma (SPA) is a rare neoplasm occurring in the salivary glands, mainly the parotid gland. Although it was originally thought to represent a non-neoplastic process, recent genetic data have proven its monoclonality, supporting its neoplastic origin. We report a case of a 73-year-old woman who presented with left neck swelling and pain. A 3 cm hypoechoic, heterogeneous, solid mass was identified on neck ultrasonography within the left parotid gland. Fine needle aspiration revealed benign acinar cells and lymphocytes. Left partial superficial parotidectomy was performed and a diagnosis of SPA was made. Targeted next-generation sequencing (NGS) revealed three clinically significant alterations in the PIK3R1, HRAS, and AR genes. Alterations in the PIK3R1 gene have been previously reported in cases of SPA; however, this study is the first to report two novel clinically significant genomic alterations in the HRAS and AR genes. AR protein expression by immunohistochemistry was strongly and diffusely positive in the neoplastic epithelial cells compared to the adjacent normal salivary gland tissue, which was dead negative for AR. This molecular profile will enhance our understanding of the molecular pathways underlying the development of this tumor. Although this entity was initially thought to be a reactive process, evidence from our case and similar cases strongly support the notion that it is neoplastic due to the presence of specific genetic alterations linked to it

Evidencias fotográfica, biológica y genética de la presencia actual de jaguaroundi (Puma yagouaroundi) en Michoacán, México Photographic, biological and genetic evidences of the presence of jaguaroundi (Puma yagouaroundi) at the moment in Michoacán, Mexico

El jaguaroundi, a pesar de su amplia distribución neotropical, es uno de los felinos menos estudiados del continente y se carece de estudios genéticos sobre la especie. Para el estado de Michoacán ha existido la sospecha de su presencia y no obstante que sólo se tenía un registro del año 1970, los mapas de distribución de la especie en México incluyen al estado. Combinando métodos de campo (trampas cámara, recolección de campo, transectos) y genotipificación molecular, obtuvimos evidencia fotográfica, biológica y genética que confirma la presencia actual de jaguaroundi (Puma yagouaroundi) en 3 regiones del estado de Michoacán, México. Se obtuvieron 11 registros fotográficos en 7 localidades con bosque tropical y 7 de estos registros, revelaron que la especie está activa principalmente por la tarde, que existen 2 fases de pelaje, predominando la fase clara y que se reproduce en el estado. Con base en las distancias e independencia entre registros de los municipios de Arteaga y Lázaro Cárdenas, se plantea la hipótesis de que la distribución continúa a lo largo de la sierra Madre del Sur y la costa del Pacífico de Michoacán, aunque se desconoce si hay conectividad hacia la depresión del Balsas. Se obtuvieron 2 secuencias de 1089 y 1096 pb del gen de citocromo b que actualmente son las más largas que se han obtenido para la especie en México y el norte del continente. Las secuencias indican que hay 2 haplotipos distintos. La presencia de la especie en 3 regiones y los 2 haplotipos permiten suponer que en Michoacán puede contar con importante diversidad genética, aunque hace falta ampliar el tamaño de muestra para confirmarlo. Las secuencias obtenidas permitirán la comparación con individuos de otras regiones del país para conocer mejor la variabilidad genética en la especie y auxiliarán en la identificación de poblaciones para conservación.The jaguaroundi is one of the least studied felids on the American continent, despite its wide neotropical distribution. Genetic studies concerning the species are also inexistent. For the state of Michoacán, it has always been assumed as present, since the distribution maps for the species in Mexico include the state, but only one record existed from 1970. Combining survey methods (camera traps, skulls and tissue found in the field, and transects) and molecular genotyping, we obtained photographic, biological and genetic evidence that confirms the actual presence of the jaguaroundi (Puma yagouaroundi) in 3 regions of Michoacán, Mexico. Eleven records were obtained from 7 localities that present tropical forests. Seven photographic records revealed that the species main activity period is during the afternoon, that both pelage phases occur in the state with a greater proportion in the clear pelage phase, and that breeding activity occurs in the state. Based on the distance and independence among Arteaga and Lázaro Cárdenas records, we hypothesize a continuous distribution of a population along the Sierra Madre del Sur and Pacific coast of Michoacán. We are unaware if a continuous distribution occurs as well along the Balsas basin. 1 089 and 1 096 pb cytochrome b gene sequences were obtained and constitute the longest sequences reported for the species in Mexico and to the north of the continent. The sequences also revealed the presence of 2 distinct haplotypes. The presence of species in 3 regions and the presence of 2 haplotypes allow us to hipothezise that in Michoacán the species may possess important genetic diversity, although a greater sample size is required for confirmation. The sequences obtained will allow the comparison with individuals from other regions of the country in order to increase the knowledge on the species genetic variability, and will provide support for the identification of populations of conservation interest

Association of ERAP2 polymorphisms in Colombian HLA-B27+ or HLA-B15+ patients with SpA and its relationship with clinical presentation: axial or peripheral predominance

Objective: To determine the association between endoplasmic reticulum aminopeptidase (ERAP)1 and ERAP2 single-nucleotide polymorphisms (SNPs) and human leukocyte antigens (HLA)-B27+ or HLA-B15+ patients with spondyloarthritis (SpA). Methods: 104 patients with SpA according to Assessment of Spondyloarthritis International Society criteria were included in the study. HLA typing was performed by PCR. The polymorphisms were determined by real-time PCR on genomic DNA using customised probes for SNPs rs27044, rs17482078, rs10050860 and rs30187 in ERAP1, and rs2910686, rs2248374 and rs2549782 in ERAP2. Results: 70 of the104 patients with SpA were HLA-B27+ and 34 were HLA-B15+. The distribution of ERAP1 and ERAP2 SNPs between the HLA-B15+ and HLA-B27+ patients with SpA did not reveal differences. Likewise, no differences in the frequencies of ERAP1 SNP haplotypes and alleles HLA-B15 or HLA-B27 were found. Interestingly, however, the frequencies of three particular haplotypes formed by ERAP2 SNPs rs2549782/rs2248374/rs2910686 varied between HLA-B15+ and HLA-B27+ patients: the ERAP2 SNPs haplotype TGT was more common in HLA-B15+ patients with SpA (OR 2.943, 95/100 CI 1.264 to 6.585; P=0.009), whereas the ERAP2 SNP haplotypes TGC and CAT were more associated with HLA-B27+ patients with SpA: (OR 4.483, 95/100 CI 1.524 to 13.187; p=0.003) and (OR 9.014, 95/100 CI 1.181 to 68.807; p=0.009), respectively. Conclusion: An association was found between HLA-B15+ patients with SpA and haplotype TGT of ERAP2 SNPs. On the other hand, HLA-B27+ patients with SpA were associated with ERAP2 haplotypes TGC and CAT. These associations could be related to the clinical presentation of the disease, specifically with a peripheral or axial predominance, respectively