The Map3k12 (Dlk)/JNK3 signaling pathway is required for pancreatic beta-cell proliferation during postnatal development.

Unveiling the key pathways underlying postnatal beta-cell proliferation can be instrumental to decipher the mechanisms of beta-cell mass plasticity to increased physiological demand of insulin during weight gain and pregnancy. Using transcriptome and global Serine Threonine Kinase activity (STK) analyses of islets from newborn (10 days old) and adult rats, we found that highly proliferative neonatal rat islet cells display a substantially elevated activity of the mitogen activated protein 3 kinase 12, also called dual leucine zipper-bearing kinase (Dlk). As a key upstream component of the c-Jun amino terminal kinase (Jnk) pathway, Dlk overexpression was associated with increased Jnk3 activity and was mainly localized in the beta-cell cytoplasm. We provide the evidence that Dlk associates with and activates Jnk3, and that this cascade stimulates the expression of Ccnd1 and Ccnd2, two essential cyclins controlling postnatal beta-cell replication. Silencing of Dlk or of Jnk3 in neonatal islet cells dramatically hampered primary beta-cell replication and the expression of the two cyclins. Moreover, the expression of Dlk, Jnk3, Ccnd1 and Ccnd2 was induced in high replicative islet beta cells from ob/ob mice during weight gain, and from pregnant female rats. In human islets from non-diabetic obese individuals, DLK expression was also cytoplasmic and the rise of the mRNA level was associated with an increase of JNK3, CCND1 and CCND2 mRNA levels, when compared to islets from lean and obese patients with diabetes. In conclusion, we find that activation of Jnk3 signalling by Dlk could be a key mechanism for adapting islet beta-cell mass during postnatal development and weight gain.

Contribution of the Long Noncoding RNA H19 to ß-Cell Mass Expansion in Neonatal and Adult Rodents.

Pancreatic ß-cell expansion throughout the neonatal period is essential to generate the appropriate mass of insulin-secreting cells required to maintain blood glucose homeostasis later in life. Hence, defects in this process can predispose to diabetes development during adulthood. Global profiling of transcripts in pancreatic islets of newborn and adult rats revealed that the transcription factor E2F1 controls expression of the long noncoding RNA H19, which is profoundly downregulated during the postnatal period. H19 silencing decreased ß-cell expansion in newborns, whereas its re-expression promoted proliferation of ß-cells in adults via a mechanism involving the microRNA let-7 and the activation of Akt. The offspring of rats fed a low-protein diet during gestation and lactation display a small ß-cell mass and an increased risk of developing diabetes during adulthood. We found that the islets of newborn rats born to dams fed a low-protein diet express lower levels of H19 than those born to dams that did not eat a low-protein diet. Moreover, we observed that H19 expression increases in islets of obese mice under conditions of increased insulin demand. Our data suggest that the long noncoding RNA H19 plays an important role in postnatal ß-cell mass expansion in rats and contributes to the mechanisms compensating for insulin resistance in obesity.

Diagnosis of subclinical atherosclerosis in HIV-infected patients: higher accuracy of the D:A:D risk equation over Framingham and SCORE algorithms.

AIMS: While the detection of subclinical atherosclerosis may provide an opportunity for the prevention of cardiovascular disease (CVD), which currently is a leading cause of death in HIV-infected subjects, its diagnosis is a clinical challenge. We aimed to compare the agreement and diagnostic performance of Framingham, SCORE and D:A:D equations for the recognition of subclinical atherosclerosis in HIV patients and to adjust the D:A:D equation using HIV and CVD variables. METHODS AND RESULTS: Atherosclerosis was evaluated in 203 HIV-infected individuals by measuring the carotid intima-media thickness (IMT). The CVD risk was calculated using the Framingham, SCORE and D:A:D risk equations. Framingham, SCORE and D:A:D equations showed a low agreement with the IMT (Kappa: 0.219, 0.298, 0.244, respectively; p = 0.743) and a moderate predictive performance, (area under the curve [AUC] = 0.686, 0.665 and 0.716, respectively; p = 0.048), with the D:A:D equation being the most accurate. Atherosclerosis was demonstrated in a significant proportion of subjects with low predicted CVD risk by all three algorithms (16.3%, 17.2%, 17.2%, respectively; p = 0.743). In patients with an estimated low CVD risk atherosclerosis was associated with older age (p = 0.012) and low CD4 counts (p = 0.021). A model was developed to adjust the D:A:D equation; a significant increase in accuracy was obtained when CD4 counts and low-grade albuminuria were included (AUC = 0.772; p < 0.001). CONCLUSION: The D:A:D equation overperforms Framingham and SCORE in HIV patients. However, all three equations underestimate the presence of subclinical atherosclerosis in this population. The accuracy of the D:A:D equation improves when CD4 counts and low-grade albuminuria are incorporated into the equation.

Liver fibrosis progression in HIV-HCV-coinfected patients treated with distinct antiretroviral drugs and impact of pegylated interferon/ribavirin therapy.

BACKGROUND: Advanced liver fibrosis frequently develops in patients with chronic hepatitis C coinfected with HIV. Non-invasive techniques for staging liver fibrosis, such as transient elastometry, may allow both periodic monitoring and examination of large patient populations. METHODS: A programme of liver fibrosis assessment using transient elastometry has been ongoing at our institution since 2004. All HIV-HCV-coinfected patients having ≥2 examinations separated by >18 months were included. Liver fibrosis progression (LFP) was defined as an increase in liver stiffness from <9.5 kPa (Metavir F0-F2) to >9.5 kPa (Metavir F3-F4), or an increase >30% in patients with baseline Metavir F3-F4. RESULTS: A total of 545 HIV-HCV-coinfected patients were analysed (mean age 41 years, 71% male, 81% intravenous drug users, mean body mass index 23.3 kg/m(2), 4.2% hepatitis B surface antigen-positive, 8.4% alcohol abuse, mean CD4(+) T-cell count 519 cells/µl). At baseline, 527 patients were on antiretroviral therapy, with the most frequent third drug being atazanavir (19.7%), efavirenz (15.9%), lopinavir (13.1%) or nevirapine (7.2%). A total of 99 (18%) patients experienced LFP during a mean (sd) follow-up of 70.9 (15.7) months. Use of protease inhibitors (OR 4.93, 95% CI 1.73, 14.0; P=0.03) and male gender (OR 5.12, 95% CI 1.37, 19.1; P=0.01) were associated with LFP. By contrast, the achievement of HCV clearance following pegylated interferon/ribavirin (PEG-IFN/RBV) therapy (OR 0.27, 95% CI 0.1, 0.79; P=0.02) was protective. Lopinavir exposure was significantly associated with LFP (OR 1.02, 95% CI 1.0, 1.04; P=0.03), whereas nevirapine was protective (OR 0.94, 95% CI 0.9, 0.99; P=0.02). CONCLUSIONS: The use of protease inhibitors, mainly lopinavir, is associated with increased LFP in HIV-HCV-coinfected patients. By contrast, nevirapine therapy and, particularly, HCV clearance with PEG-IFN/RBV significantly reduce LFP.

Triple combination therapy for hepatitis C with telaprevir exhibits greater early antiviral activity than with boceprevir.

BACKGROUND: Achievement of early viral suppression is important in patients with chronic HCV infection treated with telaprevir (TLV) or boceprevir (BOC) to avoid selection of drug resistance and attain cure. No head-to-head studies comparing TLV and BOC have been performed so far. METHODS: All consecutive individuals who initiated triple HCV therapy with TLV or BOC outside clinical trials at three European clinics were evaluated. Rapid virological response (RVR) was defined as unquantifiable HCV RNA (<25 IU/ml) at week 4 for TLV and at week 8 for BOC (4 weeks after lead-in). RESULTS: A total of 106 patients were evaluated, 33 treated with BOC and 73 with TLV. Median age, gender, body mass index, baseline HCV RNA, HCV subtype 1a (45% versus 42%) and IL28B-CC alleles (29% versus 23%) did not differ significantly in BOC and TLV groups, respectively. HIV coinfection was more prevalent in patients on TLV than BOC (24% versus 44%). Conversely, more patients on BOC than TLV had previously failed pegylated interferon plus ribavirin (82% versus 64%). RVR was achieved by 82% of patients on TLV versus 59% on BOC (P=0.001). Multivariate logistic regression analysis confirmed that TLV use was the strongest predictor of RVR (OR 3.54 [95% CI 1.23, 10.24]; P=0.02), others being HCV subtype 1b versus 1a (OR 3.26 [95% CI 1.17, 9.09]; P=0.02) and low baseline HCV RNA (OR 0.41 [95% CI 0.16, 1.03]; P=0.06). Prior interferon exposure, HIV coinfection or absence of advanced liver fibrosis did not influence the likelihood of RVR. CONCLUSIONS: Compared to BOC, triple therapy with TLV produces greater RVR rates. TLV might be a better option in more difficult-to-cure patients, such as those with high baseline HCV RNA and/or HCV 1a subtype. HIV coinfection does not influence early HCV RNA responses.

Nuevos antivirales frente al VHC: actualidad y perspectivas / New direct acting antiviral for hepatitis C: future perspectives

Aproximadamente 175 millones de personas están infectadas por el virus de la hepatitis C (VHC), lo que representa un 3% de la población mundial. En ausencia de tratamiento eficaz, un 25% de los pacientes desarrollan complicaciones hepáticas tras 25 años de hepatitis crónica C. Hasta hace poco, la única opción terapéutica en estos pacientes era la combinación de interferón pegilado (peg-IFN) y ribavirina (RBV). Alcanzaban la erradicación del VHC un 30-40% de los pacientes infectados con el genotipo 1 del VHC. Recientes avances han permitido desarrollar replicones y sistemas de cultivo tisulares para el VHC. Esto ha facilitado el diseño de fármacos antivirales directos (DAA) que inhiben específicamente la replicación del VHC. Los dos primeros inhibidores de la proteasa del VHC fueron aprobados en mayo de 2011. Permiten obtener tasas de curación en el 70% de los pacientes infectados con el genotipo 1 sin experiencia previa a interferón. La respuesta es menor en pacientes con fracasos previos, excepto en los recidivantes, en los que tasa de curación es del 90%...

Approximately 175 million people worldwide are chronically infected with the hepatitis C virus (HCV), representing 3% of the total world population. In the absence of successful therapy nearly 25% of these patients will develop hepatic complications within 25 years. Until recently, the only available therapeutic option for these patients was the combination of peginterferon-a plus ribavirin. Overall it allowed achievement of eradication in only 30-40% of patients infected by HCV genotype 1. The development of HCV replicons and the chance of producing infectious viral particles in culture systems have both enabled the rational design of direct-acting antivirals (DAA) that specifically inhibit HCV replication. The first two HCV protease inhibitors were marketed in May 2011. Triple therapy has increased the response rate to 70% in HCV genotype 1 carrier naïve to interferon. Although response rates are lower in prior failures, 90% sustained virological response rates are achieved in prior relapsers...

Liver stiffness predicts liver-related complications and mortality in HIV patients with chronic hepatitis C on antiretroviral therapy.

BACKGROUND: Liver disease is currently one of the leading causes of death in HIV individuals. Hepatic fibrosis largely mediates this effect and infection with hepatitis C virus (HCV) is the most common cause. Few studies have examined so far the predictive value of liver fibrosis staging on mortality and liver decompensation in HIV/HCV-coinfected patients. METHODS: A prospective programme of liver fibrosis assessment using transient elastometry has been ongoing at our institution since 2004. Data from all HIV/HCV-coinfected patients who underwent a transient elastometry examination and have at least 18 months of follow-up were selected for the current analysis. RESULTS: A total of 545 HIV/HCV-coinfected patients were examined (mean age 41 years, 71% men, 81% IDUs, mean BMI 23.3 kg/m2, HBsAg+ 4.2%, alcohol abuse 8.4%, mean CD4 cell count 519 cells/µl). The mean follow-up was 70.9 ± 15.7 months. During follow-up, 12 patients (2.2%) died, four of them due to hepatic complications. Liver-related events (ascites, encephalopathy, oesophageal varices or hepatocellular carcinoma) appeared in 53 patients (10%). In the multivariate analysis, baseline liver stiffness was the strongest predictor of liver-related complications [odds ratio (OR) 1.12, 95% confidence interval (CI) 1.08-1.16, P < 0.0001] and of all-cause mortality (OR 1.09, 95% CI 1.01-1.19, P = 0.02). The achievement of sustained virological response following peginterferon/ribavirin therapy during the study period was protective against the development of liver-related events (OR 0.02, 95% CI 0-0.23, P = 0.01). CONCLUSION: Liver fibrosis staging, as measured by transient elastometry, predicts liver-related complications and all-cause mortality in HIV/HCV-coinfected patients on antiretroviral therapy.

Factores clínicos y biomarcadores asociados a lesión vascular subclínica en la infección por el virus de la inmunodeficiencia humana / Clinical factors and biomarkers associated with subclinical atherosclerosis in the human immunodeficiency virus infection

Fundamento y objetivos: Los pacientes con VIH presentan aterosclerosis acelerada y la enfermedad cardiovascular es una de las principales causas de mortalidad de estos pacientes. Nuestro objetivo fue identificar biomarcadores y factores clínicos relacionados con la aterosclerosis carotídea en pacientes con VIH. Pacientes y método: Estudio transversal en pacientes con VIH. Se determinó el grosor íntima-media carotídeo (GIMc) y diferentes biomarcadores séricos en 235 sujetos. Los pacientes con GIMc≥p75 y/o placa fueron diagnosticados de enfermedad vascular subclínica (EVS). Resultados: Edad 46 (11) años. El GIMc medio fue 0,58 (0,13)mm. Sesenta y cinco (27,8%) pacientes presentaron EVS. En comparación con los pacientes sin EVS, estos mostraron mayor frecuencia de lipodistrofia (OR:2,7; IC95%:1,4-4,9) e inmunodepresión (OR:2,5; IC95%:1,1-5,8), mayor tiempo de diagnóstico del VIH (≥p50 [10 años], OR:1,4; IC95%:1,1-2,9), exposición a análogos de nucleósidos (≥p50 [132 meses], OR:3,2; IC95%:1,7-6) e inhibidores de proteasa (≥p50 [24 meses], OR:2,2; IC95%:1,1-3,6). Mostraron mayores niveles de diferentes biomarcadores séricos, destacando el NT-proBNP (≥p75 [72,6pg/ml], OR:2,0; IC95%:1,0-4,1) y el cociente albúmina/creatinina en orina (≥p50 [5mg/g], OR:3,8; IC95%:1,3-11). Tras el análisis multivariado, la EVS se relacionó con la edad (≥p50 [46 años], (OR:6,6; IC95%: 2,2-19,5; p=0,001), el tiempo de diagnóstico del VIH (≥p50 [10 años], OR:3,1; IC95%:1,0-11,0; p=0,044) y la inmunodepresión (OR:2,8, IC95%:1-8,3; p=0,048). Conclusiones: En pacientes con VIH, la edad, el tiempo de evolución de la infección y la inmunodepresión se relacionan independientemente con la EVS. Los pacientes con EVS presentaron niveles aumentados de biomarcadores de daño vascular, destacando el cociente albúmina/creatinina en orina y el NT-proBNP (AU)

Background and objective: HIV-infected patients present accelerated cardiovascular disease (CVD) and CVD is among the most important causes of mortality in this population. We aimed to identify biomarkers and clinical factors associated with subclinical atherosclerosis in HIV-infected patients. Patients and methods: Carotid intima-media thickness (cIMT) and cardiovascular biomarkers were measured in 235 HIV-infected patients. Individuals with a cIMT≥75th percentile or plaque were classified as having subclinical atherosclerosis and compared with patients without subclinical atherosclerosis. Results: Age was 46 (11) years old. Mean cIMT was 0.58 (0.13)mm. Sixty-five (27.8%) patients had subclinical atherosclerosis. These subjects had more frequently lipodystrophy (OR:2.7; CI95%:1.4-4.9), immunosuppression (OR:2.5; CI95%:1.1-5.8), longer time to HIV diagnosis (≥p50 [10 years], OR:1.4; CI95%:1.1-2.9), longer exposure to nucleoside analogues (≥p50 [132 months], OR:3.2; CI95%:1.7-6) and to protease inhibitors (≥p50 [24 months], OR:2.2; CI95%:1.1-3.6). They also showed higher levels of several biomarkers such as NT-proBNP (≥p75 [72.6pg/ml], OR:2.0; CI95%:1-4.1) and albumin/creatinine urine ratio (≥p50 [5mg/g], OR:3.8; CI95%:1.3-11). After the multivariate analysis, subclinical atherosclerosis was associated with age (OR:6.6; CI95%:2.2-19.5; P=.001), a longer time to HIV diagnosis (OR:3.1; CI95%:1.0-11.0; P=.044) and immunosuppression (OR:2.8; CI95%:1-8.3; P=.048).Conclusions: Among HIV-infected patients, time to HIV diagnosis and immunosuppression were independently associated with subclinical atherosclerosis. Patients with subclinical atherosclerosis showed increased levels of vascular damage biomarkers, especially albumin/creatinine urine ratio and NT-proBNP (AU)

[Clinical factors and biomarkers associated with subclinical atherosclerosis in the human immunodeficiency virus infection]. / Factores clínicos y biomarcadores asociados a lesión vascular subclínica en la infección por el virus de la inmunodeficiencia humana.

BACKGROUND AND OBJECTIVE: HIV-infected patients present accelerated cardiovascular disease (CVD) and CVD is among the most important causes of mortality in this population. We aimed to identify biomarkers and clinical factors associated with subclinical atherosclerosis in HIV-infected patients. PATIENTS AND METHODS: Carotid intima-media thickness (cIMT) and cardiovascular biomarkers were measured in 235 HIV-infected patients. Individuals with a cIMT ≥ 75th percentile or plaque were classified as having subclinical atherosclerosis and compared with patients without subclinical atherosclerosis. RESULTS: Age was 46 (11) years old. Mean cIMT was 0.58 (0.13)mm. Sixty-five (27.8%) patients had subclinical atherosclerosis. These subjects had more frequently lipodystrophy (OR:2.7; CI95%:1.4-4.9), immunosuppression (OR:2.5; CI95%:1.1-5.8), longer time to HIV diagnosis (≥ p50 [10 years], OR:1.4; CI95%:1.1-2.9), longer exposure to nucleoside analogues (≥ p50 [132 months], OR:3.2; CI95%:1.7-6) and to protease inhibitors (≥ p50 [24 months], OR:2.2; CI95%:1.1-3.6). They also showed higher levels of several biomarkers such as NT-proBNP (≥ p75 [72.6 pg/ml], OR:2.0; CI95%:1-4.1) and albumin/creatinine urine ratio (≥ p50 [5mg/g], OR:3.8; CI95%:1.3-11). After the multivariate analysis, subclinical atherosclerosis was associated with age (OR:6.6; CI95%:2.2-19.5; P=.001), a longer time to HIV diagnosis (OR:3.1; CI95%:1.0-11.0; P=.044) and immunosuppression (OR:2.8; CI95%:1-8.3; P=.048). CONCLUSIONS: Among HIV-infected patients, time to HIV diagnosis and immunosuppression were independently associated with subclinical atherosclerosis. Patients with subclinical atherosclerosis showed increased levels of vascular damage biomarkers, especially albumin/creatinine urine ratio and NT-proBNP.

Incipient renal impairment as a predictor of subclinical atherosclerosis in HIV-infected patients.

INTRODUCTION: HIV-infected patients present increased incidence of cardiovascular disease (CVD). Although incipient kidney function impairment has been associated with CVD in the general population, this association has not been properly addressed in HIV-infected patients. We assessed the relationship between incipient renal impairment (IRI) and subclinical atherosclerosis in HIV-infected patients. METHODS: Estimated glomerular filtration rate (eGFR), carotid intima-media thickness (cIMT), and cardiovascular biomarkers were measured in 145 HIV-infected patients. IRI was defined as a composite variable: eGFR <90 mL/min, rate of eGFR decrease >3% annually over a period of 3 years, and albumin/creatinine urine ratio above the median (≥5 mg/g). Individuals with a cIMT ≥75th percentile or plaque were classified as having subclinical atherosclerosis. RESULTS: Ninety-five patients (64.1%) met the criteria for IRI. As for HIV-related factors, patients with IRI more frequently had lipodystrophy (41.3% vs. 21.6%; P = 0.017), a lower CD4 lymphocyte nadir [210 (125-343) vs. 302 (178-408) cells/mL; P = 0.046], and longer exposure to nucleoside reverse transcriptase inhibitors [187 (84-259) vs. 104 (34-170) months; P = 0.001], to nonnucleoside reverse transcriptase inhibitors [32 (7-77) vs. 20 0-40) months; P = 0.043], and to protease inhibitors [42 (0-115] vs. 2.5 (0-59) months; P = 0.007]. Patients with IRI more frequently had subclinical atherosclerosis (40.7% vs. 13.7%; odds ratio: 4.3; 95% confidence interval: 1.8 to 10.6; P = 0.001), even after adjustment for cardiovascular and HIV-related parameters (odds ratio: 3.8; 95% confidence interval: 1.3 to 11; P = 0.012). CONCLUSIONS: The presence of IRI is an independent predictor of increased cIMT in HIV-infected patients and may help to identify patients with subclinical atherosclerosis and, therefore, increased risk of CVD.

Interacciones medicamentosas de etravirina / Etravirine drug interactions

Etravirina (ETR) es un fármaco perteneciente a la familia de los inhibidores de la transcriptasa inversa no análogos de nucleósidos (ITINAN), con actividad antiviral en situaciones de resistencia a los ITINAN de primera generación. Las interacciones farmacológicas producidas por ETR se deben a su efecto dual sobre el sistema CYP450. Es inductor de la actividad de CYP3A4 e inhibidor de la de CYP2C9 y CYP2C19. ETR presenta escasas interacciones farmacológicas clínicamente significativas, entre las que destacan los inhibidores de la proteasa sin potenciar, los ITINAN efavirenz y nevirapina, ritonavir a dosis plena y tipranavir/ritonavir. La interacción con fosamprenavir/ritonavir no es clínicamente significativa, aunque hay una escasa variación de sus valores plasmáticos al administrarse de manera conjunta con ETR. No presenta interacciones con darunavir/ritonavir

Etravirine (ETR) belongs to the family of non-nucleoside analogue reverse transcriptase inhibitors (NNRTIs), with antiviral activity in patients with resistance to first-generation NNRTIs. The drug interactions caused by ETR are due to its dual effect on the CYP450 system. ETR acts as an inducer of CYP3A4 and inhibitor of CYP2C9 and CYP2C19. This drug shows few clinically significant drug interactions, the most important of which involve the unboosted protease inhibitors, the NNRTIs efavirenz and nevirapine, full-dose ritonavir and tipranavir/ritonavir. Interaction with fosamprenavir/ritonavir is not clinically significant, although their plasma levels vary slightly when used in combination with ETR. ETR shows no interactions with darunavir/ritonavir

[Etravirine drug interactions]. / Interacciones medicamentosas de etravirina.

Etravirine (ETR) belongs to the family of non-nucleoside analogue reverse transcriptase inhibitors (NNRTIs), with antiviral activity in patients with resistance to first-generation NNRTIs. The drug interactions caused by ETR are due to its dual effect on the CYP450 system. ETR acts as an inducer of CYP3A4 and inhibitor of CYP2C9 and CYP2C19. This drug shows few clinically significant drug interactions, the most important of which involve the unboosted protease inhibitors, the NNRTIs efavirenz and nevirapine, full-dose ritonavir and tipranavir/ritonavir. Interaction with fosamprenavir/ritonavir is not clinically significant, although their plasma levels vary slightly when used in combination with ETR. ETR shows no interactions with darunavir/ritonavir.