Authorizing Third-Party Applications Served through Messaging Platforms.

The widespread adoption of smartphones and the new-generation wireless networks have changed the way that people interact among themselves and with their environment. The use of messaging platforms, such as WhatsApp, has become deeply ingrained in peoples' lives, and many digital services have started to be delivered using these communication channels. In this work, we propose a new OAuth grant type to be used when the interaction between the resource owner and the client takes place through a messaging platform. This new grant type firstly allows the authorization server to be sure that no Man-in-the-Middle risk exists between the resource owner and the client before issuing an access token. Secondly, it allows the authorization server to interact with the resource owner through the same user-agent already being used to interact with the client, i.e., the messaging platform, which is expected to improve the overall user experience of the authorization process. To verify this assumption, we conducted a usability study in which subjects were required to perform the full authorization process using both the standard authorization code grant type (through a web-browser) and the new grant type defined in this work. They have also been required to fill in a small questionnaire including some demographic information and their impressions about both authorization flows. The results suggest that the proposed grant type eases the authorization process in most cases.

Long-Term Outcomes of 496 Anatomical Cementless Modular Femoral Stems: Eleven to Twenty Years of Follow-Up.

BACKGROUND: The aim of the study is to assess the long-term outcomes of this specific stem (anatomical cementless modular stem ESOP), to review the survivorship, complication rate, and radiographic and clinical outcomes. METHODS: Descriptive and analytical retrospective longitudinal observational study of patients was operated on total hip arthroplasty between 1998 and 2007. Four hundred ninety-six prostheses corresponding to 447 patients were reviewed, mean age was 65.8 years (standard deviation [SD] ±11.6 years), and median follow-up time was 13.4 years (range 1-20). The most used cups were cementless (75.8%). The most frequent friction pairs were metal-polyethylene (53.1%) and ceramic-polyethylene (24.2%). Main variables analyzed were stem survival, subsidence, coronal orientation, osteolysis, reintervention, and Oxford Hip Score. RESULTS: From 496 implants, there were 22 lost to follow-up (4.4%). Stem revision was performed in 51 patients: 26 periprosthetic joint infections (2-stage revision), 16 periprosthetic fractures, and 8 one-stage revisions (6 real aseptic loosening with negative culture after revision). The stem survivorship at more than 15 years for any reason was 89.2% and for aseptic loosening 97.97%. No specific complications were found due to modularity. The mean subsidence and orientation was 2.06 mm (SD ±5.11 mm) and 0.41° varus (SD ±2.20°) respectively. Subsidence >5 mm or varus >5° was associated with a higher revision rate. Osteolysis was found in 110 patients (zone I = 79, VII = 57), associated with zirconium-polyethylene, without relationship to the stem revision rate or Oxford Hip Score. CONCLUSION: To our knowledge, this study represents the largest series of this stem, revealing an excellent survival rate and long-term clinical outcomes similar to the best results of classical cementless stems published in the literature.

Microservice chatbot architecture for chronic patient support.

Chatbots are able to provide support to patients suffering from very different conditions. Patients with chronic diseases or comorbidities could benefit the most from chatbots which can keep track of their condition, provide specific information, encourage adherence to medication, etc. To perform these functions, chatbots need a suitable underlying software architecture. In this paper, we introduce a chatbot architecture for chronic patient support grounded on three pillars: scalability by means of microservices, standard data sharing models through HL7 FHIR and standard conversation modeling using AIML. We also propose an innovative automation mechanism to convert FHIR resources into AIML files, thus facilitating the interaction and data gathering of medical and personal information that ends up in patient health records. To align the way people interact with each other using messaging platforms with the chatbot architecture, we propose these very same channels for the chatbot-patient interaction, paying special attention to security and privacy issues. Finally, we present a monitored-data study performed in different chronic diseases, and we present a prototype implementation tailored for one specific chronic disease, psoriasis, showing how this new architecture allows the change, the addition or the improvement of different parts of the chatbot in a dynamic and flexible way, providing a substantial improvement in the development of chatbots used as virtual assistants for chronic patients.

Marcaje de leucocitos con 99mTc-exametazima y policitemia vera: un hallazgo casual / Leucocyte labelling with 99mTc-exametazime and polycythemia vera: a casual finding

La clasificación oficial de la Organización Mundial de la Salud de los tumores de tejidos hematopoyéticos y linfoides de 2016 introduce el concepto de policitemia vera (PV) enmascarada y revela que esta entidad ha sido infradiagnosticada en el pasado. Se presenta el caso de un varón de 49 años, fumador, intervenido de fractura de tobillo hace más de 15 años, remitido para valorar un posible proceso infeccioso asociado. Al no producirse separación previa de los hematíes por sedimentación durante el procedimiento de marcaje de leucocitos con 99mTc-exametazima se revisaron las causas de disminución de la velocidad de sedimentación globular. Entre ellas destacan la poliglobulia y el hábito tabáquico, ambas presentes en el paciente. Se recomendó realización de estudio hematológico que concluyó con el diagnóstico de PV. Las indicaciones del especialista en Radiofarmacia permitieron diagnosticar un caso no identificado hasta entonces, pese a que el paciente presentaba síntomas desde hacía años

The official World Health Organization classification of hematopoietic and lymphoid tissue tumors introduces in 2016 the concept of masked polycythemia vera (PV) and reveals that this entity has been underdiagnosed in the past. We present the case of a 49-year-old man, smoker, operated on for ankle fracture more than 15 years ago, referred to evaluate a possible associated infectious process. As there was no previous separation of the red blood cells by sedimentation during the leukocyte labelling procedure with 99mTc-exametazima, the causes of decreased erythrocyte sedimentation rate were reviewed. These include polyglobulia and smoking, both present in the patient. A haematological study was advised, which concluded with the diagnosis of PV. The indications of the specialist in Radiopharmacy allowed diagnosing a case not identified until then, although the patient had had symptoms for years

Utilización de hematíes marcados con 99Mtc y desnaturalizados por calor en el diagnóstico gammagráfico de un caso infrecuente de esplenosis intratorácica / Use of heat-denatured 99mTC -labeled red blood cells in the scintigraphic diagnosis of an infrequent case of intrathoracic splenosis

La esplenosis intratorácica es poco frecuente y se asocia con historia previa de ruptura del bazo y del diafragma causado por un traumatismo. Suele ser asintomática, presentándose como un hallazgo accidental en las imágenes radiográficas o de tomografía computarizada. El diagnóstico definitivo puede realizarse mediante estudios gammagráficos asociados con estudios funcionales de captación de partículas o células. Por su sensibilidad y especificidad, la gammagrafía con hematíes marcados con 99mTc y desnaturalizados por calor es la técnica de referencia que permite confirmar el diagnóstico de esplenosis y diferenciarla de otros procesos que requieren resección quirúrgica. Se describe el caso de un varón de 52 años atendido por dolor de tipo pleurítico en hemitórax izquierdo. Las imágenes mostraron derrame pleural izquierdo e infarto pulmonar sin signos de tromboembolismo. Se evidenciaron múltiples focos sugestivos de esplenosis, que fue confirmada mediante gammagrafía esplénica con hematíes marcados con 99mTc y desnaturalizados por calor

Intrathoracic splenosis is extremely rare and is associated with previous history of rupture of the spleen and diaphragm caused by trauma. It is usually asymptomatic, presenting as an accidental finding in the X-ray images or computed tomography. The definitive diagnosis can be made by scintigraphic studies associated with functional studies of particle or cell uptake. Due to its sensitivity and specificity, gammagraphy with heat-denatured 99mTc-labeled red blood cells is the reference technique for confirming the diagnosis of splenosis and differentiating it from other processes that require surgical resection. We describe the case of a 52-year-old man treated for pleuritic pain in the left hemithorax. The images showed left pleural effusion and pulmonary infarction without signs of thromboembolism. There were multiple foci suggestive of splenosis, which was confirmed by splenic scintigraphy with heat-denatured 99mTc-labeled red blood cells

Validation of the Apple Watch for Heart Rate Variability Measurements during Relax and Mental Stress in Healthy Subjects.

Heart rate variability (HRV) analysis is a noninvasive tool widely used to assess autonomic nervous system state. The market for wearable devices that measure the heart rate has grown exponentially, as well as their potential use for healthcare and wellbeing applications. Still, there is a lack of validation of these devices. In particular, this work aims to validate the Apple Watch in terms of HRV derived from the RR interval series provided by the device, both in temporal (HRM (mean heart rate), SDNN, RMSSD and pNN50) and frequency (low and high frequency powers, LF and HF) domain. For this purpose, a database of 20 healthy volunteers subjected to relax and a mild cognitive stress was used. First, RR interval series provided by Apple Watch were validated using as reference the RR interval series provided by a Polar H7 using Bland-Altman plots and reliability and agreement coefficients. Then, HRV parameters derived from both RR interval series were compared and their ability to identify autonomic nervous system (ANS) response to mild cognitive stress was studied. Apple Watch measurements presented very good reliability and agreement (>0.9). RR interval series provided by Apple Watch contain gaps due to missing RR interval values (on average, 5 gaps per recording, lasting 6.5 s per gap). Temporal HRV indices were not significantly affected by the gaps. However, they produced a significant decrease in the LF and HF power. Despite these differences, HRV indices derived from the Apple Watch RR interval series were able to reflect changes induced by a mild mental stress, showing a significant decrease of HF power as well as RMSSD in stress with respect to relax, suggesting the potential use of HRV measurements derived from Apple Watch for stress monitoring.

Biometric Authentication Using the PPG: A Long-Term Feasibility Study.

The photoplethysmogram (PPG) is a biomedical signal that can be used to estimate volumetric blood flow changes in the peripheral circulation. During the past few years, several works have been published in order to assess the potential for PPGs to be used in biometric authentication systems, but results are inconclusive. In this paper we perform an analysis of the feasibility of using the PPG as a realistic biometric alternative in the long term. Several feature extractors (based on the time domain and the Karhunen⁻Loève transform) and matching metrics (Manhattan and Euclidean distances) have been tested using four different PPG databases (PRRB, MIMIC-II, Berry, and Nonin). We show that the false match rate (FMR) and false non-match rate (FNMR) values remain constant in different time instances for a selected threshold, which is essential for using the PPG for biometric authentication purposes. On the other hand, obtained equal error rate (EER) values for signals recorded during the same session range from 1.0% for high-quality signals recorded in controlled conditions to 8% for those recorded in conditions closer to real-world scenarios. Moreover, in certain scenarios, EER values rise up to 23.2% for signals recorded over different days, signaling that performance degradation could take place with time.

Terutroban, a TP-receptor antagonist, reduces portal pressure in cirrhotic rats.

UNLABELLED: Increased production of vasoconstrictive prostanoids, such as thromboxane A2 (TXA2 ), contributes to endothelial dysfunction and increased hepatic vascular tone in cirrhosis. TXA2 induces vasoconstriction by way of activation of the thromboxane-A2 /prostaglandin-endoperoxide (TP) receptor. This study investigated whether terutroban, a specific TP receptor blocker, decreases hepatic vascular tone and portal pressure in rats with cirrhosis due to carbon tetrachloride (CCl4 ) or bile duct ligation (BDL). Hepatic and systemic hemodynamics, endothelial dysfunction, liver fibrosis, hepatic Rho-kinase activity (a marker of hepatic stellate cell contraction), and the endothelial nitric oxide synthase (eNOS) signaling pathway were measured in CCl4 and BDL cirrhotic rats treated with terutroban (30 mg/kg/day) or its vehicle for 2 weeks. Terutroban reduced portal pressure in both models without producing significant changes in portal blood flow, suggesting a reduction in hepatic vascular resistance. Terutroban did not significantly change arterial pressure in CCl4 -cirrhotic rats but decreased it significantly in BDL-cirrhotic rats. In livers from CCl4 and BDL-cirrhotic terutroban-treated rats, endothelial dysfunction was improved and Rho-kinase activity was significantly reduced. In CCl4 -cirrhotic rats, terutroban reduced liver fibrosis and decreased alpha smooth muscle actin (α-SMA), collagen-I, and transforming growth factor beta messenger RNA (mRNA) expression without significant changes in the eNOS pathway. In contrast, no change in liver fibrosis was observed in BDL-cirrhotic rats but an increase in the eNOS pathway. CONCLUSION: Our data indicate that TP-receptor blockade with terutroban decreases portal pressure in cirrhosis. This effect is due to decreased hepatic resistance, which in CCl4 -cirrhotic rats was linked to decreased hepatic fibrosis, but not in BDL rats, in which the main mediator appeared to be an enhanced eNOS-dependent vasodilatation, which was not liver-selective, as it was associated with decreased arterial pressure. The potential use of terutroban for portal hypertension requires further investigation.

The transcription factor KLF2 mediates hepatic endothelial protection and paracrine endothelial-stellate cell deactivation induced by statins.

BACKGROUND & AIMS: Statins improve hepatic endothelial function and liver fibrosis in experimental models of cirrhosis, thus they have been proposed as therapeutic options to ameliorate portal hypertension syndrome. The transcription factor Kruppel-like factor 2 (KLF2) may be induced by statins in liver sinusoidal endothelial cells (SEC), orchestrating an efficient vasoprotective response. The present study aimed at characterizing whether KLF2 mediates statins-derived hepatic protection. METHODS: Expression of KLF2 and its vasoprotective target genes was determined in SEC freshly isolated from control or CCl(4)-cirrhotic rats treated with four different statins (atorvastatin, mevastatin, simvastatin, and lovastatin), in the presence of mevalonate (or vehicle), under static or controlled shear stress conditions. KLF2-derived vasoprotective transcriptional programs were analyzed in SEC transfected with siRNA for KLF2 or siRNA-control, and incubated with simvastatin. Paracrine effects of SEC highly-expressing KLF2 on the activation status of rat and human hepatic stellate cells (HSC) were evaluated. RESULTS: Statins administration to SEC induced significant upregulation of KLF2 expression. KLF2 upregulation was observed after 6h of treatment and was accompanied by induction of its vasoprotective programs. Simvastatin vasoprotection was inhibited in the presence of mevalonate, and was magnified in cells cultured under physiological shear stress conditions. Statin-dependent induction of vasoprotective genes was not observed when KLF2 expression was muted with siRNA. SEC overexpressing KLF2 induced quiescence of HSC through a KLF2-nitric oxide-guanylate cyclase-mediated paracrine mechanism. CONCLUSIONS: Upregulation of hepatic endothelial KLF2-derived transcriptional programs by statins confers vasoprotection and stellate cells deactivation, reinforcing the therapeutic potential of these drugs for liver diseases that course with endothelial dysfunction.

Recombinant human manganese superoxide dismutase reduces liver fibrosis and portal pressure in CCl4-cirrhotic rats.

BACKGROUND & AIMS: High oxidative stress plays a major role in increasing hepatic vascular resistance in cirrhosis, by facilitating liver fibrosis and by increasing hepatic vascular tone. This study is aimed at investigating whether the use of the novel isoform of recombinant human manganese superoxide dismutase (rMnSOD) could be a new therapeutic strategy to reduce oxidative stress and portal hypertension in cirrhotic rats. METHODS: In CCl(4)- and BDL-cirrhotic rats treated with rMnSOD (i.p. 15 µg/kg/day) or its vehicle for 7 days, mean arterial pressure (MAP), portal pressure (PP) and portal blood flow (PBF) or small mesenteric arterial flow (SMABF) were measured. In addition, in CCl(4)-cirrhotic rats, we evaluated the hepatic vasodilatory response to acetylcholine, liver fibrosis with Sirius red staining and hepatic stellate cell activation by α-smooth muscle actin (α-SMA) protein expression. RESULTS: rMnSOD treatment significantly reduced PP either in CCl(4)- or BDL-cirrhotic rats without significant changes in splanchnic blood flow, suggesting a reduction in hepatic vascular resistance. MAP was not modified. Reduction in PP was associated with a significant reduction in liver fibrosis, and α-SMA protein expression as well as with improved vasodilatory response to acetylcholine. CONCLUSIONS: Chronic rMnSOD administration to cirrhotic rats reduces portal pressure by reducing hepatic vascular resistance without deleterious effects on systemic hemodynamics, suggesting that it might constitute a new antioxidant to be considered as additional therapy for treating portal hypertension in cirrhosis.

Resveratrol improves intrahepatic endothelial dysfunction and reduces hepatic fibrosis and portal pressure in cirrhotic rats.

BACKGROUND & AIMS: Resveratrol, a polyphenol found in a variety of fruits, exerts a wide range of beneficial effects on the endothelium, regulates multiple vasoactive substances and decreases oxidative stress, factors involved in the pathophysiology of portal hypertension. Our study aimed at evaluating the effects of resveratrol on hepatic and systemic hemodynamics, hepatic endothelial dysfunction, and hepatic fibrosis in CCl4 cirrhotic rats. METHODS: Resveratrol (10 and 20 mg/kg/day) or its vehicle was administered to cirrhotic rats for two weeks and hepatic and systemic hemodynamics were measured. Moreover, we evaluated endothelial function by dose-relaxation curves to acetylcholine, hepatic NO bioavailability and TXA2 production. We also evaluated liver fibrosis by Sirius Red staining of liver sections, collagen-1, NFκB, TGFß mRNA expression, and desmin and α-smooth muscle actin (α-SMA) protein expression, as a surrogate of hepatic stellate cell activation. RESULTS: Resveratrol administration significantly decreased portal pressure compared to vehicle (12.1 ± 0.9 vs. 14.3 ± 2.2 mmHg; p <0.05) without significant changes in systemic hemodynamics. Reduction in portal pressure was associated with an improved vasodilatory response to acetylcholine, with decreased TXA2 production, increased endothelial NO, and with a significant reduction in liver fibrosis. The decrease in hepatic fibrosis was associated with a reduced collagen-1, TGFß, NFκB mRNA expression and desmin and α-SMA protein expression. CONCLUSIONS: Resveratrol administration reduces portal pressure, hepatic stellate cell activation and liver fibrosis, and improves hepatic endothelial dysfunction in cirrhotic rats, suggesting it may be a useful dietary supplement in the treatment of portal hypertension in patients with cirrhosis.

Effects of simvastatin administration on rodents with lipopolysaccharide-induced liver microvascular dysfunction.

UNLABELLED: Endothelial dysfunction drives vascular derangement and organ failure associated with sepsis. However, the consequences of sepsis on liver sinusoidal endothelial function are largely unknown. Statins might improve microvascular dysfunction in sepsis. The present study explores liver vascular abnormalities and the effects of statins in a rat model of endotoxemia. For this purpose, lipopolysaccharide (LPS) or saline was given to: (1) rats treated with placebo; (2) rats treated with simvastatin (25 mg/kg, orally), given at 3 and 23 hours after LPS/saline challenge; (3) rats treated with simvastatin (25 mg/kg/24 h, orally) from 3 days before LPS/saline injection. Livers were isolated and perfused and sinusoidal endothelial function was explored by testing the vasodilation of the liver circulation to increasing concentrations of acetylcholine. The phosphorylated endothelial nitric oxide synthase (PeNOS)/endothelial nitric oxide synthase (eNOS) ratio was measured as a marker of eNOS activation. LPS administration induced an increase in baseline portal perfusion pressure and a decrease in vasodilation to acetylcholine (sinusoidal endothelial dysfunction). This was associated with reduced eNOS phosphorylation and liver inflammation. Simvastatin after LPS challenge did not prevent the increase in baseline portal perfusion pressure, but attenuated the development of sinusoidal endothelial dysfunction. Treatment with simvastatin from 3 days before LPS prevented the increase in baseline perfusion pressure and totally normalized the vasodilating response of the liver vasculature to acetylcholine and reduced liver inflammation. Both protocols of treatment restored a physiologic PeNOS/eNOS ratio. CONCLUSION: LPS administration induces intrahepatic endothelial dysfunction that might be prevented by simvastatin, suggesting that statins might have potential for liver protection during endotoxemia.

Sinusoidal endothelial dysfunction precedes inflammation and fibrosis in a model of NAFLD.

Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome. Most morbidity associated with the metabolic syndrome is related to vascular complications, in which endothelial dysfunction is a major pathogenic factor. However, whether NAFLD is associated with endothelial dysfunction within the hepatic vasculature is unknown. The aims of this study were to explore, in a model of diet-induced overweight that expresses most features of the metabolic syndrome, whether early NAFLD is associated with liver endothelial dysfunction. Wistar Kyoto rats were fed a cafeteria diet (CafD; 65% of fat, mostly saturated) or a control diet (CD) for 1 month. CafD rats developed features of the metabolic syndrome (overweight, arterial hypertension, hypertryglyceridemia, hyperglucemia and insulin resistance) and liver steatosis without inflammation or fibrosis. CafD rats had a significantly higher in vivo hepatic vascular resistance than CD. In liver perfusion livers from CafD rats had an increased portal perfusion pressure and decreased endothelium-dependent vasodilation. This was associated with a decreased Akt-dependent eNOS phosphorylation and NOS activity. In summary, we demonstrate in a rat model of the metabolic syndrome that shows features of NAFLD, that liver endothelial dysfunction occurs before the development of fibrosis or inflammation.

Interaction between NO and COX pathways modulating hepatic endothelial cells from control and cirrhotic rats.

Reduced intrahepatic nitric oxide (NO) bioavailability and increased cyclooxygenase-1 (COX-1)-derived vasoconstrictor prostanoids modulate the hepatic vascular tone in cirrhosis. We aimed at investigating the reciprocal interactions between NO and COX in the hepatic endothelium of control and cirrhotic rats. NO bioavailability (DAF-FM-DA staining), superoxide (O(2)(-)) content (DHE staining), prostanoid production (PGI(2) and TXA(2) by enzyme immunoassays) as well as COX expression (Western Blot), were determined in hepatic endothelial cells (HEC) from control and cirrhotic rats submitted to different experimental conditions: COX activation, COX inhibition, NO activation and NO inhibition. In control and cirrhotic HEC, COX activation with arachidonic acid reduced NO bioavailability and increased O(2)(-) levels. These effects were abolished by pre-treating HEC with the COX inhibitor indomethacin. In control, but not in cirrhotic HEC, scavenging of O(2)(-) by superoxide dismutase (SOD) incubation partially restored the decrease in NO bioavailability promoted by COX activation. NO supplementation produced a significant and parallel reduction in PGI(2) and TXA(2) production in control HEC, whereas it only reduced TXA(2) production in cirrhotic HEC. By contrast, in control and cirrhotic HEC, NO inhibition did not modify COX expression or activity. Our results demonstrate that NO and COX systems are closely interrelated in HEC. This is especially relevant in cirrhotic HEC where COX inhibition increases NO bioavailability and NO supplementation induces a reduction in TXA(2). These strategies may have beneficial effects ameliorating the vasoconstrictor/vasodilator imbalance of the intrahepatic circulation of cirrhotic livers.

Addition of simvastatin to cold storage solution prevents endothelial dysfunction in explanted rat livers.

UNLABELLED: Pathophysiological alterations in the endothelial phenotype result in endothelial dysfunction. Flow cessation, occurring during organ procurement for transplantation, triggers the endothelial dysfunction characteristic of ischemia/reperfusion injury, partly due to a reduction in the expression of the vasoprotective transcription factor Kruppel-like Factor 2 (KLF2). We aimed at (1) characterizing the effects of flow cessation and cold storage on hepatic endothelial phenotype, and (2) ascertaining if the consequences of cold stasis on the hepatic endothelium can be pharmacologically modulated, improving liver graft function. Expression of KLF2 and its vasoprotective programs was determined in (i) hepatic endothelial cells (HEC) incubated under cold storage conditions with or without the KLF2-inducer simvastatin, and (ii) rat livers not cold stored or preserved in cold University of Wisconsin solution (UWS) supplemented with simvastatin or its vehicle. In addition, upon warm reperfusion hepatic vascular resistance, endothelial function, nitric oxide vasodilator pathway, apoptosis, inflammation, and liver injury were evaluated in not cold stored livers or livers preserved in cold UWS supplemented with simvastatin or vehicle. Expression of KLF2 and its vasoprotective programs decrease in HEC incubated under cold storage conditions. Cold-stored rat livers exhibit a time-dependent decrease in KLF2 and its target genes, liver injury, increased hepatic vascular resistance, and endothelial dysfunction. The addition of simvastatin to the storage solution, maintained KLF2-dependent vasoprotective programs, prevented liver damage, inflammation, and oxidative stress and improved endothelial dysfunction. CONCLUSION: Our results provide a rationale to evaluate the beneficial effects of a vasoprotective preservation solution on human liver procurement for transplantation.

Endothelial expression of transcription factor Kruppel-like factor 2 and its vasoprotective target genes in the normal and cirrhotic rat liver.

OBJECTIVE: The transcription factor Kruppel-like factor 2 (KLF2) modulates the expression of multiple endothelial vasoprotective genes. In the absence of KLF2, the endothelial phenotype becomes dysfunctional. To date, blood-derived shear stress is the main physiological stimulus identified to trigger and sustain endothelial KLF2 expression. Portal hypertension is a common complication of cirrhosis. Sinusoidal distortion and endothelial dysfunction play a significant role in its pathogenesis. This study aimed to assess whether abnormal intrahepatic haemodynamics in cirrhosis could modify KLF2 expression and consequently its downstream transcriptional programmes. DESIGN: Rats received carbon tetrachloride or vehicle for two (acute injury), six (early cirrhosis) and twelve weeks (advanced cirrhosis). Systemic and hepatic haemodynamic parameters were measured in vivo. Hepatic expression of KLF2 and its vasoprotective targets were determined. Additionally, KLF2 expression was determined in liver sections, in freshly-isolated hepatic endothelial cells, and in livers from simvastatin-treated cirrhotic animals. RESULTS: Cirrhotic livers have increased endothelial KLF2 expression compared with controls. KLF2 elevation, observed at six weeks of cirrhosis induction, was accompanied by a parallel increase in portal pressure and an increase in the expression of its target genes eNOS, thrombomodulin and CNP. Simvastatin administration further increased hepatic KLF2 and target genes expression. CONCLUSIONS: This study shows an increase in the expression of the vasoprotective transcription factor KLF2 in the cirrhotic liver, accompanied by an activation of its downstream transcriptional programmes. These data suggest that the marked increase in KLF2 expression may represent an endothelial compensatory mechanism to improve the ongoing vascular dysfunction in the cirrhotic liver.

Tempol administration, a superoxide dismutase mimetic, reduces hepatic vascular resistance and portal pressure in cirrhotic rats.

BACKGROUND & AIMS: Increased superoxide in cirrhotic livers, by reducing nitric oxide bioavailability, contributes to increase intrahepatic vascular resistance to portal blood flow and as a consequence portal pressure. We aimed to evaluate whether a strategy directed to reduce superoxide using tempol, a small membrane permeable SOD-mimetic, is able to modulate intrahepatic nitric oxide content and reduce portal pressure in cirrhotic rats. METHODS: Superoxide and nitric oxide were evaluated in control sinusoidal endothelial cells (SEC) pre-treated with the pro-oxidant diethyldithiocarbamate (DDC) and in CCl(4)-cirrhotic rat livers treated with tempol or vehicle. Mean arterial pressure, portal pressure, and portal blood flow were measured in control and cirrhotic rats treated with tempol (180µmol/kg/h; via ileocholic vein) or vehicle. In a subset of animals, hemodynamic measurements were performed after NO-inhibition with l-NAME. RESULTS: Tempol reduced superoxide content and increased NO both in SEC and cirrhotic livers. In cirrhotic rats, but not in controls, tempol significantly reduced portal pressure, and increased portal blood flow, which most likely reflects a reduction in intrahepatic vascular resistance. Tempol significantly reduced mean arterial pressure. l-NAME prevented all these effects. CONCLUSIONS: Tempol reduces superoxide, increases nitric oxide, and reduces portal pressure in sinusoidal endothelial cells and in cirrhotic livers. These results confirm that oxidative stress has a role in the pathogenesis of portal hypertension and supports the use of antioxidants in its treatment. However, when considering the use of antioxidants as additional therapy to treat portal hypertension, the potential to produce deleterious effects on systemic hemodynamics needs to be carefully evaluated.

Flow cessation triggers endothelial dysfunction during organ cold storage conditions: strategies for pharmacologic intervention.

BACKGROUND: Vascular pathologies constitute a major cause of graft rejection after organ transplantation. Recent studies have documented an improvement in transplant outcome when organs are preserved through pulsatile perfusion; however, the underlying mechanisms of these observations are poorly characterized. We hypothesized that the temporary absence of flow occurring in the context of organ cold storage conditions disrupts endothelial vasoprotective programs, and that this consequence of stasis may be a target for pharmacological modulation. METHODS: The expression of the transcription factor Kruppel-like factor 2 (KLF2) and its vasoprotective target genes were assessed during cold storage conditions in cultured human endothelial cells and murine aortic segments. In addition, we evaluated the effect of simvastatin used as a supplement in a cold preservation solution on the expression of vasoprotective genes, and on endothelial activation and apoptosis. RESULTS: The expression of endothelial KLF2 and its vasoprotective transcriptional targets were rapidly lost during cold preservation in vitro and ex vivo. Importantly, simvastatin treatment blocked the decay of KLF2, sustaining a vasoprotective phenotype, and preventing endothelial activation and apoptosis. CONCLUSIONS: Flow stasis leads to acute endothelial dysfunction and apoptosis in the context of cold storage conditions. Supplementation of organ preservation solutions with pharmacologic agents that restore endothelial vasoprotective programs, by upregulating KLF2, may represent a significant advancement of current organ preservation techniques.