Soluble urokinase plasminogen activator receptor levels are predictive of COVID-19 severity in Afro-Caribbean patients.

Aim: To investigate association between soluble urokinase plasminogen activator receptor (suPAR) plasma levels at admission and incidence of complications in COVID-19 patients. Patients & methods: We considered Afro-Caribbean patients (n = 64) admitted to the hospital between 1 February 2020 and 28 February 2021. Primary outcome was time from the hospital admission until intensive care unit care or death. Results: Primary outcome (hazard ratio, HR [95%CI]) was associated with higher CT scan severity score (3.18 [1.15-8.78], p = 0.025), National Early Warning Score (NEWS2; 1.43 [1.02-2.02], p = 0.041) and suPAR (1.28 [1.06-2.06], p = 0.041). Kaplan-Meier analysis indicated patients with suPAR level above 8.95 ng/ml had a worse outcome (7.95 [3.33-18.97], p < 0.001). Conclusion: Our study suggests that COVID-19 patients with increased baseline suPAR levels are at a high risk of complications.

Plain language summary Our aim was to investigate association between the plasma levels of soluble urokinase plasminogen activator receptor (suPAR) at admission and incidence of complications in COVID-19 patients. Increased suPAR level has been previously associated with activation of inflammation and coagulation, which important features of COVID-19. We considered Afro-Caribbean patients admitted to the hospital between 1 February 2020 and 28 February 2021. Primary outcome was time from the hospital admission until intensive care unit care or death. The use of an integrative prediction tool which combines simple clinical score (NEWS2), imaging technique (chest CT severity score) and suPAR plasma levels has potent predictive value for COVID-19 outcome.

Immunohistochemical study of the plasminogen activator system in benign epithelial odontogenic lesions

Abstract: The aim of this study was to analyze and compare the immunohistochemical expression of plasminogen activator system (PAS) proteins (uPA, uPAR, and PAI-1) in ameloblastomas (AMBs), odontogenic keratocysts (OKCs), and dental follicles (DFs) representing normal odontogenic tissue, as well as to investigate possible correlations between these proteins. Twenty AMBs, 20 OKCs, and 10 DFs were selected for immunohistochemical analysis. In each case, the immunoexpression of uPA, uPAR, and PAI-1 was evaluated semiquantitatively based on the percentage of positivity in odontogenic epithelial and connective tissue cells. The epithelial immunoexpression of uPA was significantly lower in AMBs when compared to OKCs (p = 0.001) and DFs (p = 0.029). Significantly higher epithelial immunostaining for uPAR was observed in AMBs when compared to OKCs (p < 0.001). There were no significant differences in the epithelial immunoexpression of PAI-1 between AMBs and OKCs (p = 1.000). The correlations found for the expression of the studied proteins were not statistically significant (p > 0.05). However, the epithelial and connective tissue expressions of uPAR have a strong positive and statistically significant correlation in AMBs. The present results suggest that uPA is involved in the pathogenesis of OKCs and that uPAR may participate in tumorigenesis in AMBs. The high percentage of PAI-1-positive cells suggests a possible role for this protein in the development of AMBs and OKCs. Furthermore, the studied proteins do not seem to act synergistically in AMBs, OKCs, and DFs.

Gene therapy with urokinase-type plasminogen activator regenerates liver tissue.

BACKGROUND AND OBJECTIVES: Gene therapy using urokinase-type plasminogen activator (uPA) has been shown to induce extracellular matrix degradation, hepatocyte proliferation and liver tissue function restoration in liver cirrhosis models. Physiologically, uPA activates plasminogen conversion to plasmin, which leads, depending on the organ, to thrombolysis or extracellular matrix degradation. The purpose of this study was to compare the regenerative effect of gene therapy with adenoviruses encoding wild-type uPA (huPA), as well as its truncated isoform (ΔhuPA), in healthy and cirrhotic animals. In addition, possible adverse effects on coagulation were assessed. METHODS: 6 x 1011 vp/kg of Ad-huPA or Ad-ΔhuPA were administered via the iliac vein to healthy male Wistar rats or to male Wistar rats with cirrhosis induced by chronic poisoning with carbon tetrachloride (CCl4). The animals were sacrificed at day 2, 4 or 6 post-treatment. Liver fibrosis, proliferating cell nuclear antigen expression, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) serum levels and coagulation markers were evaluated. RESULTS: On day 6 post-treatment, a fibrosis reversal of 48.7-41.5% was achieved. AST and ALT levels did not change in cirrhotic animals treated with ΔhuPA, but showed an elevation in healthy animals. Cell proliferation increased in healthy and cirrhotic animals with both transgene isoforms. No coagulation adverse effects were observed in the ΔhuPA group, and by day 6, they had disappeared in the huPA group. CONCLUSIONS: Gene therapy with Ad-huPA and Ad-ΔhuPA favors cell proliferation in cirrhotic animals, without important side effects.

ANTECEDENTES Y OBJETIVOS: La terapia génica empleando el activador de plasminógeno tipo urocinasa (huPA) ha demostrado que induce la degradación de matriz extracelular, la proliferación de hepatocitos y el restablecimiento de la funcionalidad tisular en modelos de cirrosis hepática. Fisiológicamente, el uPA activa la conversión del plasminógeno en plasmina, lo que conlleva, dependiendo del órgano, una trombólisis o a degradación de la matriz extracelular. El objetivo de este estudio fue comparar el efecto regenerador de la terapia génica con adenovirus codificando la forma silvestre (huPA), así como su isoforma truncada (DhuPA) en animales sanos y cirróticos. Además, se valoraron los posibles efectos adversos sobre la coagulación. MÉTODOS: Se administró 6x1011 pv/kg del Ad-huPA o Ad-ΔhuPA a ratas Wistar macho sanas o con cirrosis por intoxicación crónica con tetracloruro de carbono (CCl4) vía vena ilíaca. El sacrificio fue al día 2, 4 o 6 postratamiento. Se evaluó la fibrosis hepática, la expresión de antígeno nuclear de proliferación celular y niveles séricos de alanina aminotransferasa (ALT) y aspartato aminotransferasa (AST), y marcadores de coagulación. RESULTADOS: Al día 6 postratamiento se alcanzó una reversión de fibrosis del 48.7-41.5%. Los niveles de AST y ALT no cambiaron en animales cirróticos tratados con DhuPA, pero aumentaron en animales sanos comparados con el control sano no tratado. La proliferación aumenta en animales sanos y cirróticos con ambas isoformas del transgén. No se observaron efectos adversos en la coagulación en el grupo DhuPA y para el día 6 habían desaparecido en el grupo huPA. CONCLUSIONES: La terapia génica con Ad-uPA y Ad-ΔhuPA favorece la proliferación celular en animales cirróticos, sin efectos secundarios importantes.

Epidermal growth factor receptor regulates fibrinolytic pathway elements in cervical cancer: functional and prognostic implications

Epidermal growth factor receptor (EGFR) signaling and components of the fibrinolytic system, including urokinase-type plasminogen activator (uPA) and thrombomodulin (TM), have been implicated in tumor progression. In the present study, we employed cBioPortal platform (http://www.cbioportal.org/), cancer cell lines, and an in vivo model of immunocompromised mice to evaluate a possible cooperation between EGFR signaling, uPA, and TM expression/function in the context of cervical cancer. cBioPortal analysis revealed that EGFR, uPA, and TM are positively correlated in tumor samples of cervical cancer patients, showing a negative prognostic impact. Aggressive human cervical cancer cells (CASKI) presented higher gene expression levels of EGFR, uPA, and TM compared to its less aggressive counterpart (C-33A cells). EGFR induces uPA expression in CASKI cells through both PI3K-Akt and MEK1/2-ERK1/2 downstream effectors, whereas TM expression induced by EGFR was dependent on PI3K/Akt signaling alone. uPA induced cell-morphology modifications and cell migration in an EGFR-dependent and -independent manner, respectively. Finally, treatment with cetuximab reduced in vivo CASKI xenografted-tumor growth in nude mice, and decreased intratumoral uPA expression, while TM expression was unaltered. In conclusion, we showed that EGFR signaling regulated expression of the fibrinolytic system component uPA in both in vitro and in vivo settings, while uPA also participated in cell-morphology modifications and migration in a human cervical cancer model.

HO-1 Interactors Involved in the Colonization of the Bone Niche: Role of ANXA2 in Prostate Cancer Progression.

BACKGROUND: Prostate cancer (PCa) dissemination shows a tendency to develop in the bone, where heme oxygenase 1 (HO-1) plays a critical role in bone remodeling. Previously by LC/ESI-MSMS, we screened for HO-1 interacting proteins and identified annexin 2 (ANXA2). The aim of this study was to analyze the relevance of ANXA2/HO-1 in PCa and bone metastasis. METHODS: We assessed ANXA2 levels using a co-culture transwell system of PC3 cells (pre-treated or not with hemin, an HO-1 specific inducer) and the pre-osteoclastic Raw264.7 cell line. RESULTS: Under co-culture conditions, ANXA2 mRNA levels were significantly modulated in both cell lines. Immunofluorescence analysis unveiled a clear ANXA2 reduction in cell membrane immunostaining for Raw264.7 under the same conditions. This effect was supported by the detection of a decrease in Ca2+ concentration in the conditioned medium. HO-1 induction in tumor cells prevented both, the ANXA2 intracellular relocation and the decrease in Ca2+ concentration. Further, secretome analysis revealed urokinase (uPA) as a key player in the communication between osteoclast progenitors and PC3 cells. To assess the clinical significance of ANXA2/HO-1, we performed a bioinformatics analysis and identified that low expression of each gene strongly associated with poor prognosis in PCa regardless of the clinico-pathological parameters assessed. Further, these genes appear to behave in a dependent manner. CONCLUSIONS: ANXA2/HO-1 rises as a critical axis in PCa.

Estudo da imunoexpressão de proteínas do sistema ativador de plasminogênio em lesões benignas do epitélio odontogênico / Study of immunoexpression of plasminogen activator system proteins in benign lesions of the odontogenic epithelium

Introdução: Cistos e tumores odontogênicos são lesões originadas a partir de tecidos que formam os elementos dentários e apresentam diferentes comportamentos biológicos. Dentre os diversos elementos que podem estar associados ao crescimento cístico e tumoral, encontram-se as enzimas necessárias para degradação da matriz extracelular (MEC). O sistema ativador de plasminogênio (SAP) é responsável pela regulação da remodelação da MEC através da conversão do plasminogênio em plasmina. Além disto, diversos estudos têm sugerido associações entre o SAP e vários fatores na evolução de neoplasias malignas, como a transição epitélio-mesênquima, proliferação, migração, adesão celular e disseminação metastática. Entretanto, poucos trabalhos foram realizados avaliando a influência do SAP em lesões odontogênicas. Objetivo: Avaliar e comparar a expressão imuno-histoquímica das proteínas do SAP, o ativador do plasminogênio tipo uroquinase (uPA), seu receptor (uPAR) e o inibidor (PAI-1) em ameloblastomas (AMBs), ceratocistos odontogênicos (COs) e tecidos odontogênicos normais, os folículos dentários (FDs), além de investigar possíveis correlações entre as proteínas estudadas. Materiais e métodos: As células epiteliais odontogênicas foram analisadas, de forma semi-quantitativa, a partir de fotomicrografias de 5 campos representativos de cada caso, com ampliação de 400x, sendo atribuídos escores de 0 a 4 de acordo com o percentual de células positivas. Após a análise dos 5 campos, foi obtida a mediana dos escores, sendo gerado o escore de imunomarcação do caso. Os dados foram submetidos à análise estatística por meio dos testes de Kruskal-Wallis (KW), Mann-Whitney (U) e correlação de Spearman (r), com o nível de significância estabelecido em 5% (p<0,05). Resultados: A imunoexpressão de uPA foi significativamente menor em AMBs, quando comparados com COs (p=0,001) e FDs (p=0,029), enquanto que a imunomarcação de uPAR em AMBs foi significativamente maior em comparação aos COs (p<0,001). Não houve diferenças significativas na imunoexpressão do PAI-1 entre os grupos estudados (p=0,775). Também não foram encontradas correlações estatisticamente significativas entre as proteínas avaliadas (p>0,05). Conclusões: Os resultados do presente estudo sugerem que o uPA esteja envolvido no crescimento dos COs, enquanto que o uPAR participe do processo de tumorigênese dos AMBs, contudo, o PAI-1 parece não contribuir, de forma direta, na patogênese de AMBs e COs. No contexto das lesões estudadas, as proteínas do SAP parecem não atuar sinergicamente (AU).

Background: Odontogenic cysts and tumors are lesions originated from odontogenic tissues and exhibit different biological behaviors. Among the various elements that may be associated with cystic and tumor growth are the enzymes required for extracellular matrix (ECM) degradation. The plasminogen activator system (PAS) is responsible for regulating ECM remodeling by converting plasminogen to plasmin. In addition, several studies have suggested associations between PAS and other factors in the evolution of malignant neoplasms, such as epithelial-mesenchymal transition, proliferation, migration, cell adhesion, and metastatic dissemination. However, just a few studies have evaluated the influence of PAS on odontogenic lesions. Aim: To evaluate and to compare the immunohistochemical expression of PAS proteins, the urokinase-type plasminogen activator (uPA), its receptor (uPAR) and inhibitor (PAI-1) in ameloblastomas (AMBs), odontogenic keratocysts (OKCs) and normal odontogenic tissues, dental follicles (FDs), in addition to investigating possible correlations between these proteins. Materials and methods: Odontogenic epithelial cells were analyzed, in a semi-quantitative way, using photomicrographs of 5 representative fields of each case, with 400x magnification, with scores ranging from 0 to 4 according to the percentage of positive cells. After the analysis of 5 fields, the median of the scores was obtained, generating the immunostaining score of the case. The data were submitted to statistical analysis using Kruskal-Wallis (KW) and Mann-Whitney (U) tests and Spearman's correlation (r), with significance level set at 5% (p<0.05). Results: For uPA, the immunoexpression was significantly lower in AMBs, when compared to OKs (p=0.001) and DFs (p=0.029). While, uPAR immunostaining in AMBs was significantly higher compared to OKs (p<0.001). There were no significant differences in PAI-1 immunoexpression between the groups studied (p=0.775). There were also no statistically significant correlations between the evaluated proteins (p>0.05). Conclusions: The results of this study suggest that uPA is involved in growth of OKCs, while uPAR participates in tumorigenesis process of AMBs, however, PAI1 does not seem to contribute, directly, to the pathogenesis of AMBs and OKCs. In the context of the studied lesions, SAP proteins do not seem to act synergistically (AU).

Fibrin γ/γ' influences the secretion of fibrinolytic components and clot structure.

BACKGROUND: In healthy subjects fibrinogen γ/γ' circulates at 8-15% of the total plasma fibrinogen concentration. Elevated levels of this variant have been associated with arterial thrombosis, and its diminution with venous thrombosis. The aims of the present work were to analyze the structure of the fibrin network formed on the top of human dermal microvascular endothelial cells (HMEC-1) at different fibrinogen γ/γ' concentrations, as well as its influence on the secretion of fibrinolytic components. The kinetics of fibrin polymerization on top of HMEC-1 cells with 3, 10, and 30% fibrinogen γ/γ' was followed at 350 nm. The secretion of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI 1) by HMEC-1 were measured in the supernatant and cell lysates, after incubation with 1 nM thrombin, fibrin with 3, and 30% fibrinogen γ/γ', using commercial kits. The influence of fibrinogen γ/γ' on fibrin structure on the surface of the HMEC-1 was followed with laser scanning confocal microscopy (LSCM). RESULTS: The kinetics of fibrin formation on HMEC-1 with 3 and 10% fibrinogen γ/γ' were similar. However, with 30% fibrinogen γ/γ' both the slope and final turbity were approximately 50% less. The LSCM images showed the dramatic effects of increasing fibrinogen γ/γ' from 3 to 30%. The uPA and PAI 1 concentrations in culture supernatants HMEC-1 cells treated with thrombin or 30% γ/γ' fibrin were two-fold increased as compared to basal culture supernatants and 3% γ/γ' fibrin-treated HMEC-1. In all stimulatory conditions the intracellular concentration of uPA was higher than in supernatants. In contrast, the intracellular PAI 1 concentration was decreased as compared to that measured in the supernatant, including the basal condition. CONCLUSION: A concentration of 30% fibrin γ/γ' alter drastically fibrin structure on the cell surface and affects the secretion of uPA and PAI 1 through its capacity to bind thrombin.

suPAR na avaliação do prognóstico após permanência na unidade de terapia intensiva: um estudo piloto / suPAR in the assessment of post intensive care unit prognosis: a pilot study

RESUMO Objetivo: Determinar o desempenho da dosagem do receptor ativador de plasminogênio tipo uroquinase solúvel quando da alta da unidade de terapia intensiva para predição da mortalidade após permanência na mesma unidade. Métodos: Durante 24 meses conduziu-se um estudo prospectivo observacional de coorte em uma unidade de terapia intensiva polivalente de oito leitos. Colheram-se os seguintes dados: APACHE II, SOFA, níveis de proteína C-reativa e receptor ativador de plasminogênio tipo uroquinase solúvel, além de contagem de leucócitos no dia da alta da unidade de terapia intensiva, em pacientes que sobreviveram à permanência na unidade de terapia intensiva. Resultados: Durante este período, incluíram-se no estudo 202 pacientes; 29 (18,6%) morreram após alta da unidade de terapia intensiva. Os não sobreviventes eram mais idosos e tinham enfermidades mais graves quando admitidos à unidade de terapia intensiva, com escores de severidade mais elevados, e necessitaram de vasopressores por mais tempo do que os que sobreviveram. As áreas sob a curva Característica de Operação do Receptor para SOFA, APACHE II, proteína C-reativa, contagem de leucócitos e receptor ativador de plasminogênio tipo uroquinase solúvel, no momento da alta da unidade de terapia intensiva, avaliadas como marcadores de prognóstico de morte hospitalar, foram, respectivamente, 0,78 (IC95% 0,70 - 0,86); 0,70 (IC95% 0,61 - 0,79); 0,54 (IC95% 0,42 - 0,65); 0,48 (IC95% 0,36 - 0,58); 0,68 (IC95% 0,58 - 0,78). O SOFA associou-se de forma independente com risco mais elevado de morte no hospital (OR 1,673; IC95% 1,252 - 2,234), assim como para mortalidade após 28 dias (OR 1,861; IC95% 1,856 - 2,555) e mortalidade após 90 dias (OR 1,584; IC95% 1,241 - 2,022). Conclusão: A dosagem do receptor ativador de plasminogênio tipo uroquinase solúvel na alta unidade de terapia intensiva teve um valor prognóstico fraco de mortalidade após a permanência nesta unidade.

ABSTRACT Objective: To determine the performance of soluble urokinase-type plasminogen activator receptor upon intensive care unit discharge to predict post intensive care unit mortality. Methods: A prospective observational cohort study was conducted during a 24-month period in an 8-bed polyvalent intensive care unit. APACHE II, SOFA, C-reactive protein, white cell count and soluble urokinase-type plasminogen activator receptor on the day of intensive care unit discharge were collected from patients who survived intensive care unit admission. Results: Two hundred and two patients were included in this study, 29 patients (18.6%) of whom died after intensive care unit discharge. Nonsurvivors were older and more seriously ill upon intensive care unit admission with higher severity scores, and nonsurvivors required extended use of vasopressors than did survivors. The area under the receiver operating characteristics curves of SOFA, APACHE II, C-reactive protein, white cell count, and soluble urokinase-type plasminogen activator receptor at intensive care unit discharge as prognostic markers of hospital death were 0.78 (95%CI 0.70 - 0.86); 0.70 (95%CI 0.61 - 0.79); 0.54 (95%CI 0.42 - 0.65); 0.48 (95%CI 0.36 - 0.58); and 0.68 (95%CI 0.58 - 0.78), respectively. SOFA was independently associated with a higher risk of in-hospital mortality (OR 1.673; 95%CI 1.252 - 2.234), 28-day mortality (OR 1.861; 95%CI 1.856 - 2.555) and 90-day mortality (OR 1.584; 95%CI 1.241 - 2.022). Conclusion: At intensive care unit discharge, soluble urokinase-type plasminogen activator receptor is a poor predictor of post intensive care unit prognosis.

Soluble urokinase-type plasminogen activator receptor as a measure of treatment response in acute exacerbation of COPD / Receptor do ativador de plasminogênio tipo uroquinase solúvel como medida da resposta ao tratamento da exacerbação aguda da DPOC

ABSTRACT Objective: To evaluate the value of soluble urokinase-type plasminogen activator receptor (suPAR) in the diagnosis of acute exacerbation of COPD (AECOPD) and in monitoring treatment response, analyzing the relationship between suPAR and fibrinogen in AECOPD. AECOPD leads to increased airway inflammation, contributing to an exaggerated release of inflammatory mediators. Methods: We recruited 45 patients with AECOPD and 20 healthy control subjects. Medical histories were taken, and all subjects underwent clinical examination, chest X-ray, pulmonary function tests, and blood gas analysis. On day 1 (treatment initiation for the AECOPD patients) and day 14 (end of treatment), blood samples were collected for the determination of serum suPAR and plasma fibrinogen. Results: Serum levels of suPAR were significantly higher in the AECOPD group than in the control group. In the AECOPD patients, there was a significant post-treatment decrease in the mean serum suPAR level. The sensitivity, specificity, and accuracy of suPAR were 95.6%, 80.0%, and 93.0%, respectively. The Global Initiative for Chronic Obstructive Lung Disease stage (i.e., COPD severity) correlated positively and significantly with serum levels of suPAR and plasma levels of fibrinogen. Conclusions: Monitoring the serum suPAR level can be helpful in the evaluation of the COPD treatment response and might be a valuable biomarker for determining the prognosis of AECOPD. Because serum suPAR correlated with plasma fibrinogen, both markers could be predictive of AECOPD.

RESUMO Objetivo: Avaliar o valor do soluble urokinase-type plasminogen activator receptor (suPAR, receptor do ativador de plasminogênio tipo uroquinase solúvel) no diagnóstico de exacerbação aguda da DPOC (EADPOC) e no monitoramento da resposta ao tratamento, analisando-se a relação entre o suPAR e o fibrinogênio na EADPOC. A EADPOC leva ao aumento da inflamação das vias aéreas, contribuindo para a liberação exagerada de mediadores inflamatórios. Métodos: Foram recrutados 45 pacientes com EADPOC e 20 controles saudáveis. Realizou-se anamnese, e todos os indivíduos foram submetidos a exame clínico, radiografia de tórax, provas de função pulmonar e gasometria arterial. No dia 1 (início do tratamento para os pacientes com EADPOC) e no dia 14 (final do tratamento), foram coletadas amostras de sangue para dosagem de suPAR sérico e de fibrinogênio plasmático. Resultados: Os níveis séricos de suPAR foram significativamente maiores no grupo EADPOC do que no grupo controle. Nos pacientes com EADPOC, houve diminuição significativa da média de suPAR sérico após o tratamento. A sensibilidade, a especificidade e a acurácia do suPAR foram, respectivamente, de 95,6%, 80,0% e 93,0%. O estágio da doença segundo a Global Initiative for Chronic Obstructive Lung Disease (isto é, a gravidade da DPOC) apresentou correlação positiva e significativa com os níveis séricos de suPAR e os níveis plasmáticos de fibrinogênio. Conclusões: O monitoramento do suPAR sérico pode ser útil na avaliação da resposta ao tratamento da DPOC e seria um biomarcador valioso para a determinação do prognóstico da EADPOC. Como o suPAR sérico apresentou correlação com o fibrinogênio plasmático, ambos os marcadores poderiam ser preditores da EADPOC.

In IgA Nephropathy, Glomerulosclerosis Is Associated with Increased Urinary CD80 Excretion and Urokinase-Type Plasminogen Activator Receptor-Positive Podocyturia.

BACKGROUND: Podocyturia may determine the evolution to podocytopenia, glomerulosclerosis, and renal failure. According to the Oxford classification of IgA nephropathy (IgAN), the S1 lesion describes glomerulosclerosis. Urokinase-type plasminogen activator receptor (uPAR) participates in podocyte attachment, while CD80 increases in glomerulosclerosis. We measured uPAR-positive urinary podocytes and urinary CD80 (uCD80) in controls and in IgAN subjects with M1E0S0T0 and M1E0S1T0 Oxford scores to assess a potential association between podocyturia, inflammation, and glomerulosclerosis. METHODS: The groups were as follows: controls (G1), n = 20 and IgAN group (G2), n = 39, subdivided into M1E0S0T0 (G2A), n = 21 and M1E0S1T0 (G2B), n = 18. Among the included variables, we determined uPAR-positive podocytes/gram of urinary creatinine (gUrCr) and uCD80 ng/gUrCr. Biopsies with interstitial fibrosis and tubular atrophy <10% were included. RESULTS: Groups were not different in age and gender; urinary protein-creatinine (uP/C) ratio, Chronic Kidney Disease-Epidemiology Collaboration (CKD-EPI) equation, uPAR-positive podocytes/gUrCr, and uCD80 were significantly increased in G2 versus G1. G2A and G2B were not different in age, gender, hypertension, and follow-up. G2B displayed significantly higher uP/C, uPAR-positive podocytes, uCD80, and lower CKD-EPI versus G2A. Strong significant correlations were encountered between uCD80 and podocyturia in G2A and G2B. However, when G1 was compared to G2A and G2B separately, the differences with respect to uP/C, uPAR-positive podocytes, and podocyturia were significantly stronger versus G2B than versus G2A. CONCLUSIONS: IgAN presents elevated uCD80 excretion and uPAR-positive podocyturia, while CD80 correlates with podocyturia. Glomerulosclerosis (S1) at the time of biopsy is associated with higher uP/C, lower renal function, increased uPAR-positive podocyturia, and CD80 excretion, and is independent of M1. In IgAN, uPAR may participate in podocyte detachment.

Expressão imuno-histoquímica do ativador de plasminogênio do tipo uroquinase e seu receptor em carcinoma epidermoide de língua oral e sua relação com parâmetros clínico-patológicos / Immunohistochemical expression of plasminogen activator of the urokinase type and its receptor in squamous cell carcinoma of tongue and their relationship with clinicopathological parameters

O carcinoma epidermoide oral (CEO) é a neoplasia maligna mais comum da cavidade oral, apresentando uma alta taxa de mortalidade. Devido a isto, a descoberta de biomarcadores que facilitem a compreensão do comportamento biológico desse tumor e aprimorem o tratamento é necessário. O ativador de plasminogênio do tipo uroquinase (uPA) e o seu receptor, uPAR, têm se destacado por atuarem na proteólise de estruturas da membrana basal e matriz extracelular, facilitando a invasão tumoral. O presente estudo se propôs a avaliar a imunoexpressão dessas proteínas em 46 casos de carcinoma epidermoide de língua oral (CELO). Esses resultados foram relacionados com a presença de metástase, estadiamento clínico TNM, recidiva locoregional, desfecho da lesão e gradação histológica de malignidade. A imunomarcação de cada caso foi avaliada semiquantitativamente, tanto no front de invasão como no centro do tumor, na qual foram atribuídos os escores: 0 (0% de células positivas), 1(1-10% de células positivas), 2 (11-50% de células positivas), 3 (mais de 50% de células positivas). A expressão do uPA foi observada em 93,5% dos casos no front de invasão, com predomínio do escore 2 (34,8%), e em 67,9% dos casos no centro do tumor, com predomínio do escore 1 (32,6%). De modo geral, os parâmetros clínicos não exerceram influência na imunoexpressão do uPA. Em relação à gradação histológica, foi observada uma maior expressão de uPA nos casos de alto grau de malignidade em relação aos de baixo grau de malignidade (p=0,05). Quando analisado em relação aos parâmetros morfológicos, foi identificado uma maior expressão do uPA nos casos de pior padrão de invasão (p=0,03).

A expressão do uPAR foi observada em 73,9% dos casos no front de invasão, com predomínio do escore 1 (45,65%), e em 47,5% dos casos no centro do tumor, com predomínio do escore 0 (54,35%). Embora não tenham sido observadas significâncias estatísticas em relação à metástase linfonodal, estadiamento clínico TNM, desfecho e gradação histológica, houve uma maior expressão do uPAR nos casos com recidiva locoregional em relação aos sem recidiva (p=0,04). Em relação à análise da localização tumoral, foi observada uma maior expressão de uPA e uPAR no front de invasão em relação ao centro do tumor (p<0,001). Na correlação entre uPA e uPAR, não foi observada significância estatística. Com base nestes resultados, sugere-se que o uPA e uPAR estejam envolvidos na progressão do CELO, atuando principalmente na região mais profunda do tumor. (AU)

Squamous cell carcinoma (SCC ) is the most common malignancy of the oral cavity (OSCC), with a high mortality rate. Due to this, the discovery of biomarkers that facilitate the understanding of the biological behavior of the tumor and improve treatment is necessary. Urokinase type plasminogen activator (uPA) and its receptor, uPAR, are responsible for the proteolysis of structures of the basement membrana and extracellular matrix, facilitating tumor invasion. This study aims to assess the immuno expression of these proteins in 46 cases of squamous cell carcinoma of the oral tongue (OTSCC). These results were related to the presence of metastasis, clinical TNM staging, locoregional recurrence, outcome of the lesion and histological grading. Immunostaining of each case was evaluated semiquantitatively, in the front of invasion and center of the tumor, in which scores were assigned: 0 (0% of positive cells), 1 (1-10% of positive cells), 2 (11 -50% positive cells) and 3 (more than 50% positive cells). The expression of uPA was observed in 93.5% (n=43) of the cases in the front of invasion, with predominance of score 2 (n=16; 34.8%) and in 67.9% (n=31) of the cases in the center of the tumor, with predominance of score 1 (n=15; 32.6%). Overall, the immunoexpression of uPA was not associated with clinical parameters. Regarding the malignant histological grading, a higher expression of uPA was observed in cases of high-grade malignancy comp ared to low-grade malignancy (p=0.05).

Regarding the morphological parameters, increased expression of uPA was observed in the worst mode of invasion (p=0.03 ). The expression of uPAR was observed in 73.9% of cases in the front of invasion, with a predominance of score 1 (n=21; 45.6 %), and in 47.5% (n=21) of the cases in the center of the tumor, with a predominance of score 0 (n=25; 54.4%). Although no statistical differences were observed in relation to lymph node metastasis, clinical TNM staging, outcome, and histological grading, there was a higher expression of uPAR in cases with locoregional recurrence (p=0.04). Regarding the tumor intra -localization, it was observed an increased expression of uPA and uPAR at the front of invasion in relation to the center of the tumor (p<0.001). Regarding the correlation between uPA and uPAR, there was no statistical sign ificance. Based on these results, it is suggested that uPA and uPAR are involved in the progression of CELO, mainly in the deeper region of the tumor. (AU)

Alteplase vs. urokinase for occluded hemodialysis catheter: A randomized trial.

Introduction Thrombosis of tunneled central venous catheters (CVC) in hemodialysis (HD) patients is common and it can lead to the elimination of vascular sites. To compare the efficacy of alteplase vs. urokinase in reestablishing adequate blood flow through completely occluded vascular catheters. Methods In this randomized study, patients with completely occluded tunneled HD catheters received 40 minutes intracatheter dwell with alteplase (1 mg/mL) or urokinase (5000 IU/mL). Primary endpoint was the proportion of patients with occluded catheters achieving post-thrombolytic blood flow of ≥250 mL/min. Safety endpoints included the incidence of hemorrhagic and infectious complications. Findings Eligible adult patients (n = 100) were treated with alteplase (n = 44) or urokinase (n = 56). The two groups were similar in gender (male: 51.8% vs. 56.8%, P = 0.35), age (60 ± 12 vs. 59 ± 13 years, P = 0.71), time on dialysis (678 ± 203 vs. 548 ± 189 days, P = 0.77), diabetes and cardiovascular disease (55.6% vs. 70.4%, P = 0.08 and 17.8% vs. 22.7%, P = 0.38, respectively), jugular vein as main vascular access (54.8% vs. 62.5%, P = 0.57), and time of CVC (278 ± 63 vs. 218 ± 59 days, P = 0.67). Primary success with alteplase and urokinase occurred in 42/44 (95%) vs. 46/56 (82%), P = 0.06. Success was not achieved after the second dose of alteplase and urokinase in 1 and 7 cases, respectively (2% vs. 12%, P = 0.075). Serious adverse effects were not observed in both groups. There was no difference between the two groups in infectious complications (P = 0.94). Discussion Alteplase and urokinase are effective thrombolytic agents for restoring HD catheter patency. Our study has revealed a likely slight superiority of alteplase over urokinase for unblocking central lines, but which has enrolled too few patients to be able to detect a difference of this size.

Fibrinólisis como tratamiento en la trombosis de la vena subclavia: nuestra experiencia en causas primarias y secundarias / Fibrinolysis as Treatment in Subclavian Vein Thrombosis: Our Experience in Primary and Secondary Causes

La trombosis de la vena subclavia reviste especial gravedad por las secuelas funcionales y las posibles complicaciones sistémicas que puede desencadenar cuando no se diagnostica a tiempo. Aunque en muchos casos la presentación clínica puede ser diagnóstica, siempre se requieren pruebas de imagen, ya sean no invasivas, como la ecografía con Doppler color, o invasivas, como la flebografía, que es considerada el patrón de oro, ya que demuestra el trombo y confirma la permeabilidad de la circulación colateral. El objetivo de este estudio es demostrar los resultados de la fibrinólisis por catéter y comprobar que su tratamiento agresivo está justificado para evitar secuelas incapacitantes, especialmente en gente joven. Entre el 1 enero de 2006 y el 31 de diciembre de 2012 se atendieron cinco pacientes diagnosticados con trombosis en la vena subclavia, y fueron tratados con fibrinólisis endovenosa con uroquinasa a 100.000 UI/hora. El criterio para escoger los pacientes a trombolizar, fue el tiempo de evolución menor de 6 días y las características agudas del trombo (trombo hipoecogénico-homogéneo, que ocasiona aumento del calibre venoso), así como las características del paciente, edad y repercusión clínica. En nuestra limitada experiencia hemos tenido un éxito del 80 %, con resolución de la sintomatología en el 100 % en trombólisis con uroquinasa, por lo cual la recomendamos como el manejo inicial de las trombosis subclavias, siempre y cuando cumplan las indicaciones y haya ausencia de contraindicaciones absolutas o relativas para trombólisis.

Subclavian vein thrombosis can be particularly serious due to the functional consequences and possible systemic complications that can be triggered when not diagnosed early. Although in many cases the clinical presentation may be diagnostic, imaging is always required, either non-invasive (Doppler ultrasound) and / or invasive, as is the case with venography which is considered the gold standard because it shows the thrombus and confirms the permeability of the collateral circulation. The purpose of the study is to demonstrate the results of catheter directed fibrinolysis and suggests that aggressive treatment of this condition is justified to prevent the possible disabling sequelae, particularly in young people. During the period between January 1, 2006 to December 31, 2012, 5 patients were treated for thrombosis in the subclavian vein with catheter directed thrombolysis with urokinase at 100,000 UI/hour. The selection criteria for thrombolysis, was the time of evolution (less than 6 days) and acute characteristics of thrombus in ultrasound (hypo echogenic-homogenous thrombus, which causes an increase in venous caliber), as well as the characteristics of the patient, the age, and clinical repercussions. In our limited experience we had a success rate of 80% due to the dissolution of the thrombus, with a 100% resolution of symptoms in thrombolysis with urokinase; so we recommend it as the initial management of the subclavian thrombosis as long as the patients are symptomatic and have not contraindications to thrombolysis.

Inhibition of nitric oxide is a good therapeutic target for bladder tumors that express iNOS.

Bladder cancer is the second cause of death for urological tumors in man. When the tumor is nonmuscle invasive, transurethral resection is curative. On the other hand, radical cystectomy is the treatment chosen for patients with invasive tumors, but still under treatment, these patients have high risk of dying, by the development of metastatic disease within 5 years. It is therefore important to identify a new therapeutic target to avoid tumor recurrences and tumor progression. Nitric oxide (NO) is an important biological messenger known to influence several types of cancers. In bladder cancer, production of NO and expression and activity of inducible NO synthase was associated to recurrence and progression. The objective of this work was to analyze if inhibition of nitric oxide production could be considered a therapeutic target for bladder tumors expressing iNOS. Using a bladder cancer murine model with different invasiveness grade we have demonstrated that NO inhibition was able to inhibit growth of bladder tumors expressing iNOS. Furthermore, invasive properties of MB49-I orthotopic growth was inhibited using NO inhibitors. This paper also shows that levels of NO in urine can be correlated with tumor size. In conclusion, inhibition of NO could be considered as a therapeutic target that prevents tumor growth and progression. Also, urine NO levels may be useful for measuring tumor growth.

Diferencias cinéticas de las plasminas de ocho especies mamíferas: activaciones y secuencias de los terminales-N / Kinetic Differences of Plasmins from Eight Mammalian Species: N-Terminal Activations and Sequences / Diferenças cinéticas das plasminas de oito espécies mamíferas: ativações e sequencias dos terminais-N

El artículo determina los parámetros cinéticos de las plasminas de ocho especies de mamíferos y sus terminales-N. Los ocho plasminógenos fueron purificados por los mismos métodos (cromatografías de afinidad e intercambio iónico), y activados con urocinasa, partiendo de una concentración común y su cinética valorada según las coordenadas de Lineweaver-Burk. Para esto se utilizó la cinética enzimática de Michaelis Menten, ampliamente usada en el estudio de enzimas. Todos los plasminógenos mostraron una pureza superior al 95% y una banda de 92 kDa en la electroforesis, al comparar con el estándar de peso molecular utilizado. Las plasminas del equino y canino mostraron la misma K M por el sustrato cromogénico (0,438 mM), siendo esta la de mayor afinidad en este estudio y la humana la de menor afinidad (5,3 mM). También fueron determinadas la constante catalítica y la velocidad de conversión del sustrato cromogénico a producto. Los terminales-N de los plasminógenos de las ocho especies fueron determinados, y se encontraron diferencias entre el humano y los animales; así mismo entre algunos animales. Solo el porcino y el ovino no mostraron diferencia alguna en su terminal-N (DPPDDY). Se demostró la unificación del método de purificación de los plasminógenos para cualquier especie, las diferencias cinéticas de las ocho plasminas estudiadas, y las similitudes y diferencias en la secuencia de los terminales-N de las ocho especies.

The paper determines the kinetic parameters of plasmins from eight species of mammals and their N-terminals. They were purified by the same methods (affinity chromatography and ion exchange) and activated with urokinase, starting from a common concentration and its kinetics judged according to Lineweaver-Burk coordinates. For such purpose, MichaelisMenten enzyme kinetics, which is widely used in the study of enzymes, was implemented. When compared with the molecular weight standard used, all plasminogen showed a purity exceeding 95% and a 92 kDa band on electrophoresis. Equine and canine plasmins showed the same K M due to the chromogenic substrate (0.438 mM), this being the one with the highest affinity in this study, and the human being the one with the lower affinity (5.3 mM). The catalytic constant and the conversion rate of the chromogenic substrate to product were also determined. The N-terminals of the plasminogens of the eight species were determined, and differences were found between humans and animals, as well as between some animals. Only pigs and sheep showed no differences in their N-terminal (DPPDDY). The unification of the method for purification of plasminogens for any species was demonstrated, as well as the kinetic differences of the eight plasmins studied and the similarities and differences in the sequence the N-terminals of the eight species.

El artigo determina os parâmetros cinéticos das plasminas de oito espécies de mamíferos e seus terminais-N. Estes foram purificados pelos mesmos métodos (cromatografias de afinidade e intercambio iônico) e ativados com uroquinase, partindo de uma concentração comum e sua cinética avaliada segundo as coordenadas de Lineweaver-Burk. Para isto se utilizou a cinética enzimática de Michaelis-Menten, amplamente usada no estudo de enzimas. Todos os plasminogênios mostraram uma pureza superior a 95 % e uma banda de 92 kDa na eletroforese, ao comparar com o padrão de peso molecular utilizado. As plasminas do equino e canino mostraram a mesma KM pelo substrato cromogênico (0,438 mM), sendo esta a de maior afinidade neste estudo e a humana a de menor afinidade (5,3 mM). Também foram determinadas a constante catalítica e a velocidade de conversão do substrato cromogênico a produto. Os terminais-N dos plasminogênios das oito espécies foram determinados, e se encontraram diferenças entre o humano e os animais, da mesma forma entre alguns animais. Somente o suíno e o ovino não mostraram diferença alguma em seu terminal-N (DPPDDY). Demonstrou-se a unificação do método de purificação de os plasminogênios para qualquer espécie, as diferenças cinéticas das oito plasminas estudadas e as similitudes e diferencias na sequencia dos terminais-N das oito espécies.

Primary focal and segmental glomerulosclerosis and soluble factor urokinase-type plasminogen activator receptor.

Primary focal and segmental glomerulosclerosis (FSGS) may be due to genetic or acquired etiologies and is a common cause of nephrotic syndrome with high morbidity that often leads to end-stage renal failure. The different available therapeutic approaches are unsuccessful, in part due to partially deciphered heterogeneous and complex pathophysiological mechanisms. Moreover, the term FSGS, even in its primary form, comprises a histological description shared by a number of different causes with completely different molecular pathways of disease. This review focuses on the latest developments regarding the pathophysiology of primary acquired FSGS caused by soluble factor urokinase type plasminogen activator receptor, a circulating permeability factor involved in proteinuria and edema formation, and describes recent advances with potential success in therapy.

Comparación de la activación y la cinética de laplasmina bufalina con la humana / Ativação e cinética comparativa da plasmina bufalina e a humana / Comparative activation and kinetic of plasmin bufaline with the human one

El plasminógeno es el zimógeno de la plasmina, enzima activada a nivel fisiológico por el activadortisular del plasminógeno y la urokinasa, la plasmina es la enzima encargada de disolver el coágulosanguíneo. En este estudio se compararon la plasmina humana con la bufalina en su forma de activación dezimógeno a enzima y en la afinidad hacia el sustrato cromogénico. Los plasminógenos fueron purificadospor el mismo método de cromatografías de afinidad y cambio iónico. De igual manera las activaciones sehicieron utilizando urokinasa humana en ambos casos. La plasmina bufalina demostró mayor activacióny afinidad (1.35mM) que la plasmina humana (2.16 mM), siendo la bufalina 1.5 veces mas afin al sustratocromogénico que la humana. Este estudio demuestra que el método de purificación de los plasminógenospuede ser el mismo para muchas especies, se demuestra una vez más que las plasminas animales al parecerson más eficientes en la disolución del coágulo o degradación de sustratos, que la plasmina humana.Este estudio indica que la plasmina bufalina puede ser utilizada en los parámetros que se determinanclínicamente en pacientes con problemas cardiovasculares, reduciendo el tiempo de determinación de estosparámetros fibrinolíticos, que pueden dar al médico un margen de tiempo superior para actuar.

The Plasminogen is the zymogene of the Plasmin, enzyme which physiologically is activated by twodifferent enzymes, the tissue plasminogen activator and the urokinase, the plasmin is the enzyme that dissolves blood clots. In this study the human plasmin was compared to the bufaline plasmin, in theactivation from the zymogene to the enzyme form as well as in the affinity to the chromogenic substrate.The two plasminogens were purified by the same chromatographies methods: affinity and ion-exchange.Furthermore, both plasminogens were activated by human urokinase. The bufaline plasmin showed moreactivation and affinity (1.35 mM) that the human plasmin (2.16 mM), in addition, the bufaline plasmindemonstrated a 1.5 times more affinity to the chromogenic substrate that the human plasmin. This studydemonstrated that the plasminogens of several species can be purified by this method. Besides, one moretime the animal’s plasmins probably to be more efficient in the dissolution of blood clots or degradation ofsubstrates than the human plasmin. More over this study indicated that the bufaline plasmin can be usedin clinical determinations of patients with cardiovascular diseases. This also reduces the determinationtime of fibrinolytic parameters that physicians can give, having more time to take appropriate treatment.

O plasminogênio é o zymogen da plasmina, enzima ativada a nivel fisiológico pelo ativador tissulardo plasminogênio e uroquinase, plasmina é a enzima responsável de dissolver o coágulo de sanguíneo.neste estudo foi comparada a plasmina humana com a plasmina búbalina em seu modo de ativaçãode zymogen a enzima e na afinidade substrato cromogênico. Os plasminogênio foram purificados como mesmo método de cromatografia de afinidade e de troca iônica, e as ativações foram feitas usandouroquinase humana nos dois casos. A Búfalo plasmina mostrou maior ativação e afinidade (1.35 mM)que a plasmina humana (2.16 mM), sendo a bufalina 1.5 vezes mais afim ao substrato Cromogênico quea humana. Este estudo mostrou que o método de purificação do plasminogênios pode ser o mesmo paramuitas espécies, alem disso, que as plasminas animais são mais eficientes na dissolução do coáguloo degradação de substratos que a plasmina humana. Este estudo indicou que a plasmina búfalo podeser utilizada nos parâmetros determinados clínicamente em pacientes com problemas cardiovasculares,diminuindo o tempo de determinação destes parâmetros fibrinolíticos, que podem dar ao médico umintervalo de maior tempo para atuar.