Quantification of serum markers of hepatitis B (HBV) and Delta virus (HDV) infections in patients with chronic HDV infection.

The interplay between hepatitis B (HBV) and delta (HDV) viruses is complex and not always characterized during chronic HDV infection. We assessed the clinical usefulness of new quantitative assays for HBV and HDV serum markers in a retrospective cross-sectional study. Sera obtained from 122 HDV genotype 1 and HBV genotype D coinfected, anti-HIV-negative patients (71 males; median age 49.8 [21.7-66.9] years), recruited consecutively in two geographical areas (Italy 69 patients, Romania 53 patients) with different HBV and HDV epidemiology, were tested for HBsAg, HBV-DNA, HBcrAg, total anti-HBc, HDV-RNA, IgM and total anti-HDV using quantitative assays. Cirrhosis, which showed comparable prevalence in the two cohorts, was diagnosed in 97 of 122 (79.5%) patients. At multivariate analysis, cirrhosis was associated with lower total anti-HBc/IgM anti-HDV ratio (OR 0.990, 95% CI 0.981-0.999, P = .038), whereas disease activity was associated with higher total anti-HDV (OR 10.105, 95% CI 1.671-61.107, P = .012) and HDV-RNA levels (OR 2.366, 95% CI 1.456-3.844, P = .001). HDV-RNA serum levels showed a positive correlation with HBV-DNA (ρ = 0.276, P = .005), HBsAg (ρ = 0.404, P < .001) and HBcrAg (ρ = 0.332, P < .001). The combined quantitative profiling of HBV and HDV serum markers identifies specific patterns associated with activity and stage of chronic hepatitis D (CHD). HDV pathogenicity depends on the underlying active HBV infection in spite of the inhibition of its replication. HDV-RNA, IgM anti-HDV, total anti-HDV, total anti-HBc, HBsAg and HBcrAg serum levels qualify for prospective studies to predict progressive CHD and identify candidates to antiviral therapy.

Propiedades morfológicas en la parte media de la díafisis del hueso metacarpiano III de equino mestizo criollo / Morphological properties in the mid part of the metacarpal III bone diaphysis of equine criollo crossbreed

El desarrollo del esqueleto es de máxima importancia en caballos de trabajo y alta competencia. Los miembros torácicos sostienen el 60% del peso corporal y están sometidos a esfuerzos biomecánicos que los predisponen a lesiones, estando obligados a especializarse en el apoyo. El metacarpiano III soporta la mayor carga entre los huesos del cuerpo y la sustancia compacta tiene gran espesor dorsal y medial. Es un material complejo que posee plasticidad y fuerza considerable, con capacidad para soportar la deformación y recuperar su forma. El objetivo de este estudio es aportar conocimientos morfológicos, aplicables a la clínica animal, del hueso metacarpiano III de equino mestizo criollo. Se estudiaron los huesos metacarpiano III de 30 equinos hembras y machos, entre 18 meses y 5 años de edad, divididos en dos grupos etarios, provenientes del frigorífico General Pico, Las Higueras. Se determinó: peso y longitud total; diámetro latero-medial y dorso-palmar en la parte media de la diáfisis. Al metacarpiano III izquierdo se le practicó una osteotomía transversal en la parte media de la diáfisis para determinar: espesor de la cortical, área cortical, área medular y área total. Los datos fueron analizados mediante técnicas de correlación y regresión lineal simple, análisis de la varianza y comparaciones múltiples de Tukey usando el paquete estadístico InfoStat, con un nivel de significación de 0,05. Los resultados muestran que existe asociación entre el peso versus área cortical (r= 0,76, p= 0,0001) y longitud del hueso versus área cortical (r= 0,74 p= 0,0001), el área depende linealmente del peso y longitud del hueso (R2= 0,97, p<0,0001 y R2= 0,96, p<0,0001 respectivamente). Se comprobó que existen diferencias significativas entre los diámetros de las áreas y espesor de los cuadrantes (p<0,0001), al contrastar los cuadrantes entre sí se observaron diferencias a excepción del cuadrante dorsal y lateral. Existe diferencias entre las áreas de los distintos grupos etários (p= 0,0034). La morfología tiene influencia sobre las propiedades biomecánicas del hueso.

The development of high skeleton is important in working horses and associated high competition. Thoracic members hold 60% of body weight and are subjected to biomechanical efforts that predispose the animal to injury, being forced to rely on the support. The large metacarpal III supports load between bones of the body and the compact substance is large dorsal and medial thickness. It is composed of complex material and possesses plasticity and considerable force, with the ability to sustain deformation and subsequently recover its form. The objective of this study is to provide morphometric knowledge applied to animal clinic of the metacarpal III bone of equine criollo crossbreed. Thirty (30) metacarpal III bones of male and female horses, between 18 months and 5 years old, divided into two age groups, from the Frigorifico General Pico, Las Higueras groups were studied. We determined: weight, total length, diameter latero-medial and dorsal-palmar in the middle of the diaphysis. A transverse osteotomy was performed left to metacarpal III in the middle diaphysis to determine: cortical thickness, cortical area, medullary area and total area. The data was analyzed by correlation and simple linear regression techniques, analysis of variance and Tukey multiple comparisons using the statistical InfoStat (2009) package, with a significance level of 0.05. The results showed an association between weight vs. cortical area (r= 0.76, p= 0.0001) and bone length vs. cortical area (r= 0.74, p= 0.0001), the area depends linearly on the weight and length bone (R2= 0.97, p<0.0001 and R2= 0.96, p<0.0001 respectively). It was found that there are significant differences between the diameters of the area and thickness of the quadrants (p<0.0001), contrasting quadrants were observed in the dorsal and lateral exception quadrant. There are differences between areas of different age groups (p= 0.0034). The morphology influences the biomechanical properties of the bone.

HCV E1E2-MF59 vaccine in chronic hepatitis C patients treated with PEG-IFNα2a and Ribavirin: a randomized controlled trial.

Hepatitis C virus (HCV) vaccines may be able to increase viral clearance in combination with antiviral therapy. We analysed viral dynamics and HCV-specific immune response during retreatment for experienced patients in a phase Ib study with E1E2MF59 vaccine. Seventy-eight genotype 1a/1b patients [relapsers (30), partial responders (16) and nonresponders (32) to interferon-(IFN)/ribavirin-(RBV)] were randomly assigned to vaccine (V:23), Peg-IFNα2a-180-ug/qw and ribavirin 1000-1200-mg/qd for 48 weeks (P/R:25), or their combination (P/R + V:30). Vaccine (100 µg/0.5 mL) was administered intramuscularly at week 0-4-8-12-24-28-32-36. Neutralizing of binding (NOB) antibodies and lymphocyte proliferation assay (LPA) for E1E2-specific-CD4 + T cells were performed at week 0-12-16-48. Viral kinetics were analysed up to week 16. The vaccine was safe, and a sustained virological response (SVR) was achieved in 4 P/R + V and 2 P/R patients. Higher SVR rates were observed in prior relapsers (P/R + V = 27.3%; P/R = 12.5%). Higher NOB titres and LPA indexes were found at week 12 and 16 in P/R + V as compared to P/R patients (P = 0.023 and 0.025, P = 0.019 and <0.001, respectively). Among the 22 patients with the strongest direct antiviral effects of IFN (ε ≥ 0.800), those treated with P/R + V (10) reached lower HCV-RNA levels (P = 0.026) at week 16. HCV E1E2MF59 vaccine in combination with Peg-IFNα2a + RBV was safe and elicited E1E2 neutralizing antibodies and specific CD4 + T cell proliferation. Upon early response to IFN, vaccinations were associated with an enhanced second phase viral load decline. These results prompt phase II trials in combination with new antiviral therapies.

Características mecánicas dinámicas de la tibia aislada de perro sometida a prueba de impacto / Dynamic mechanical characteristics of the isolated dog tibia, subjected to impact test

El propósito del presente trabajo es brindar información acerca de las propiedades biomecánicas y morfológicas de la tibia de perro sometida a cargas dinámicas. Los objetivos fueron evaluar la resistencia de la tibia frente al impacto mediante el Método de Ensayo Charpy, relacionar la misma con el área total, el área cortical y el área de la cavidad medular del hueso, comparándola entre talla, edad y sexo diferente. La importancia clínica, ortopédica y quirúrgica que reviste la región de la pierna del perro fundamenta esta investigación. Se utilizaron tibias de perro en condiciones aisladas, extraídas postmortem de 30 animales mestizos adultos jóvenes provenientes del servicio de zoonosis de la ciudad de Río Cuarto. Se determinó en la tibia derecha e izquierda: peso y longitud total del hueso, diámetros cráneo-caudal y latero-medial en la mitad de la diáfisis. A la tibia izquierda se le practicó osteotomía transversal en la mitad de su diáfisis. En la superficie de sección se midió: espesor de la cortical, diámetro medular y se obtuvo el área de sección total, área cortical y área medular. La tibia derecha fue sometida al Método de Ensayo Charpy. Se realizó el análisis estadístico correspondiente. La resistencia del hueso presentó una correlación de r=0,70 (p= 0,000019); r=0,67 (p=0,000059) y r=0,29 (p=0,12) con respecto a las áreas cortical, total y medular respectivamente. La morfología tiene influencia sobre las propiedades biomecánicas del hueso. A medida que aumenta el área cortical aumenta la energía de impacto absorbida. En cambio, a mayor área de cavidad medular la energía de impacto absorbida aumenta pero en menor magnitud. Existen diferencias estadísticamente significativas (p=0,0168) entre la talla del animal y la resistencia de la tibia. La edad (p=0,73) y el sexo (p=0,18) no presentaron influencias significativas sobre las propiedades biomecánicas de la tibia.

The purpose of this work is to provide information about biomechanical and morphological properties of the dog tibia bone subject to dynamic loads. The objectives were: evaluate the resistance of the tibia subject impact used Charpy test Method; linking it with total area, cortical area and bone marrow cavity; and compare it with weight, age and sex categories. The clinical, orthopedical and surgical relevance of the dog leg region is the basis for this research. Dog tibia were used in isolated conditions, removed postmortem from 30 adult animals defined raceless, from the zoonoses service of the city of Río Cuarto. Weight and total bone length, cranial-caudal and lateral-medial diameter in the middle of the diaphysis, were determined in the left and right tibia. The left tibia was osteotomized transversely in the middle of the diaphysis. On the surface section we measured: cortical thickness, diameter of the bone marrow cavity, and obtained the total section area, cortical area and medullary area. The right tibia was subjected to Charpy test method. Statistical analysis was performed accordingly. The bone strength showed a correlation of r=0.70 (p= 0.000019); r=0.67 (p=0.000059) and r=0.29 (p=0.12) with respect to the cortical area, total area and medullary area respectively. The morphology has influence on the biomechanical properties of bone, with increasing cortical area increases the impact energy absorbed. In contrast, a greater area of the medullary cavity of impact energy absorbed increases but to a lesser magnitude. There were statistically significant differences (p=0.0168) of size on the strength of the tibia. Age (p=0.7343) and sex (p=0.1898) had no significant influence on the biomechanical properties of the tibia.

Multicenter evaluation of the Elecsys hepatitis B surface antigen quantitative assay.

The Elecsys hepatitis B surface antigen (HBsAg) II quantitative assay is a new quantitative electrochemiluminescence immunoassay which uses onboard dilution and a simple algorithm to determine HBsAg levels expressed in international units (IU)/ml (standardized against the World Health Organization [WHO] Second International Standard). This study evaluated its performance using routine serum samples from a wide range of HBsAg carriers and patients with chronic hepatitis B (CHB). HBsAg levels were measured in serum samples collected independently by five centers in Europe, Australia, and Asia. Serial dilution analyses were performed to assess the recommended dilution algorithm and determine the assay range free of hook effect. Assay precision was also established. Following assessment of serial dilutions (1:100 to 1:1,000,000) of the 611 samples analyzed, 70.0% and 85.6% of samples tested with analyzers incorporating 1:100 (Elecsys 2010 and cobas e 411) and 1:400 (Modular Analytics E170) onboard dilution, respectively, fell within the linear range of the assay, providing a final result on the first test. No high-dose hook effect was seen up to the maximum HBsAg serum level tested (870,000 IU/ml) using the dilution algorithm. HBsAg levels were reliably determined across all hepatitis B virus (HBV) genotypes, phases of HBV infection, and stages of disease tested. Precision was high across all analyzers (% coefficient of variation [CV], 1.4 to 9.6; HBsAg concentrations, 0.1 to 37,300 IU/ml). The Elecsys HBsAg II quantitative assay accurately and reliably quantifies HBsAg in routine clinical samples. Onboard dilution minimizes retesting and reduces the potential for error.

Características mecánicas dinámicas del fémur aislado de perro, sometido a prueba de impacto / Dynamic mechanical characteristics of the isolated dog femur, subjected to impact test

El presente trabajo tiene el propósito de brindar información acerca del comportamiento biomecánico del fémur de perro sometido a cargas dinámicas. El objetivo es evaluar la resistencia del fémur frente al impacto, mediante el Método de Ensayo Charpy, relacionando dicha resistencia con las áreas: total, cortical y cavidad medular del hueso, comparándola entre edad y sexo diferentes. Se utilizaron fémur de perro en condiciones aisladas, extraídos postmortem de 30 animales mestizos adultos, provenientes del servicio de zoonosis de la ciudad de Río Cuarto. Se determinó en los fémures derecho e izquierdo: peso y longitud total del hueso, diámetro cráneo-caudal y latero-medial en la mitad de la diáfisis. El fémur izquierdo fue osteotomizado transversalmente en la mitad de su diáfisis. En la superficie de sección se medió espesor de la cortical y diámetro medular. Se obtuvo el área de sección total, cortical y medular. El fémur del miembro pélvico derecho fue sometido al Método de Ensayo Charpy. Los resultados obtenidos fueron analizados estadísticamente encontrándose que el área cortical depende linealmente de la longitud del hueso (r=0,62, p= 0,00025); la resistencia del hueso depende de las áreas cortical (r=0,71, p= 0,000012) y área total (global) (r=0,63, p= 0,00021) y no del área medular (r=0,29, p= 0,12). El análisis exploratorio muestra que no existirían diferencias acerca de la influencia de la edad y el sexo sobre la resistencia del fémur. Tras estos resultados se puede inferir que a medida que aumenta el área cortical aumenta la energía de impacto absorbida.

This paper is intended to provide information about biomechanical behavior femur at impact using the Charpy test method relating the resistance in the following areas: total, cortical and medullary bone cavity, comparing these between different ages and sexes. Dog femors were used in isolated conditions, removed postmortem from 30 adult animals, defined raceless, from the zoonoses service of the city of Rio Cuarto. Weight, total bone length, cranio-caudal diameter and lateral medial half of the diaphysis, were determined in the left and right femur. The left femur was osteotomized transversely in the middle of the diaphysis. In the sectional area thickness of the cortical and medullary diameters were measured. We obtained the total sectional area, cortical and medullary. The femur of the right lower limb was subjected to the Charpy test method. Results were statistically analyzed and found that cortical area depends linearly on the length of the bone (r=0.62, p=0.00025), bone strength depends on the cortical areas (r=0.71, p=0.00012) and total (global) areas (r=0.63, p=0.00021) and not of the medullary area (r=0.29, p=0.12). Exploratory analysis showed that no difference existed regarding age and sex influence on the strength of the femur. Following these results we can infer that as cortical area increases, the impact energy absorbed also increases.

Quantification of copper phases, their reducibility and dispersion in doped-CuCl2/Al2O3 catalysts for ethylene oxychlorination.

The comprehensive understanding of the composition, behaviour and reactivity of a catalyst used inside industrial plants is an extremely hard task that is rarely achieved. It requires the use of different spectroscopic techniques, applied under in situ or in operando conditions, and combined with the investigation of the catalyst activity. Often the operating experimental conditions are different from technique to technique and the different results must be compared with care. In the present contribution, we combined in situ XANES/EXAFS, IR spectroscopy of adsorbed CO, CO chemisorption and catalytic tests performed using a pulse reactor in depletive mode. This multitechnical approach resulted in the understanding of the role that dopants (LiCl, KCl, CsCl, MgCl(2) LaCl(3)) have in the nature, relative fraction, reducibility and dispersion of Cu-phases on CuCl(2)/gamma-Al(2)O(3) catalysts for oxychlorination reaction, a key step of the PVC chemistry. In the undoped catalyst two Cu phases coexist: Cu-aluminate and supported CuCl(2), being the latter the only active one [J. Catal., 2000, 189, 91]. EXAFS and XANES highlighted that all dopants contribute more or less efficiently in increasing the fraction of the active copper species, that reaches a value of almost 100% in the case of MgCl(2) or LaCl(3). EXAFS directly, and IR indirectly, proved that the addition of KCl or CsCl (and less efficiently of LiCl) results in the formation of mixed CuK(x)Cl(2+x) or CuCs(x)Cl(2+x) phases, so altering the chemical nature of the active phase. XANES spectroscopy indicates that addition of MgCl(2) or LaCl(3) does not affect the reducibility by ethylene (under static conditions) of the active CuCl(2) phase and that the reducibilility of the new copper-dopant mixed chloride are in the order CuCl(2) > CuLi(x)Cl(2+x) > CuK(x)Cl(2+x) > CuCs(x)Cl(2+x). However, when reduction is done inside a pulse reactor, a more informative picture comes out. The last technique is able to differentiate all samples, and their ability to be reduced by ethylene resulted in the order: La- > Mg- > Li-doped > undoped > K- > Cs-doped catalyst. To understand this apparent discrepancy the dispersion of the active phase, measured by CO chemisorption, was needed: it has been found that addition of LiCl increases enormously the dispersion of the active phase, LaCl(3) significantly and MgCl(2) barely, while addition of both KCl and CsCl results in a decrease of the surface area of the active phase. The mechanism of the enhancing effect of La and Mg on catalytic activity is still not clear, but it could be associated to the modification that they induce to the support surface: the Cu is so highly dispersed that almost all is in direct contact with support surface. It is finally worth noticing that the previous EXAFS and XANES study allowed us to refer the chemisorption data to the active phase only, while the IR study allowed us to fix the Cu(+)/CO surface stoichiometry. Summarizing the use of a multidisciplinary approach has been the conditio sine qua non (mandatory condition) to understand the complex role that the different additives have on the active phase of the CuCl(2)/gamma-Al(2)O(3) catalysts for ethylene oxychlorination.

Micro-FTIR and micro-raman studies of a carbon film prepared from furfuryl alcohol polymerization.

The synthesis of a carbon film by the acid-catalyzed polymerization and resinification of furfuryl alcohol with a diluted solution of HCl is studied by combining micro-FTIR and micro-Raman spectroscopies. The detailed study of the evolution of spectra as a function of dosage of furfuryl alcohol and temperature shows that neutral and protonated species are formed at 80 degrees C, while upon gradually increasing the temperature up to 600 degrees C, the viscous polyfurfuryl alcohol resin is transformed into a carbon phase, containing a heterogeneous distribution of pores, with a size in the 100-2000 nm range, as shown by SEM and AFM analyses.

Análisis multivariado aplicado a la etapa de deshidratación en la técnica de plastinación del riñón de caballo / Multivaried analysis applied at the stage of dehydration in the plastination technique in horse´s kidney

La Plastinación es la técnica más moderna para la conservación de piezas anatómicas y especímenes. Esta técnica consta de cuatro etapas: fijación, deshidratación, impregnación forzada y curado. Este trabajo pretende mejorar la eficiencia en la producción de órganos plastinados mediante la técnica de Plastinación estándar denominada S 10. Se utilizaron riñones de equinos mestizos criollos provenientes del Frigorífico Aimar S/A, ubicado en la zona rural de Río Cuarto. Luego de realizar la fijación del órgano con formalina se procedió a deshidratarlo. La deshidratación se llevó a cabo en tres sub-etapas donde en cada una de ellas se trabajó a la misma temperatura y se utilizó como solvente una solución de acetona en diferentes concentraciones. Las mediciones se realizaron con acetómetro registrándose el tiempo insumido para lograr la deshidratación. Los datos se analizaron estadísticamente por medio de: análisis de la varianza multivariada y análisis de correlación simple utilizando el paquete estadístico InfoStat. Los resultados muestran que los tiempos de deshidratación se ven influenciados por la edad del animal pero no por la posición ni por el peso del órgano.

Plastination is the most modern technique for the preservation of anatomical pieces and specimens. This technique consists of four stages: fixation, dehydration, forced impregnation, and cured. This research aims at improving the efficiency of production plastinated organs through the standard technique known as S10. Kidneys of criole mestizo equines from Frigorifico Aimar S.A placed in the rural area of Rio IV were used. After binding the organ with formalin, it was dehydrated. The dehydration was carried out in three sub-stages working at the same temperature, and a solvent composed of solution of acetone of different concentrations was used. The measurements were analysed with acetometer, keeping register of the time consumption to reach dehydration. The resulting data was statistically analysed using Infostat program. Results showed that the time consumed to reach dehydration was influenced by the horses age but not by the kidney's weight, there were no significant differences between right and left kidneys neither.

Early and accurate prediction of Peg-IFNs/ribavirin therapy outcome in the individual patient with chronic hepatitis C by modeling the dynamics of the infected cells.

A novel biomathematical model that analyzes the combined alanine transaminase (ALT) and viral-load kinetics during the first month of pegylated interferon (Peg-IFN) plus ribavirin (RBV) therapy was successfully applied in 90 of 97 (93%) chronic hepatitis C patients in order to compute the number of infected cells at the end of therapy (I(eot)). The I(eot) indices were lower in sustained virological responders than in relapsers (RELs) and nonresponders (NRs) (median values: 31 vs. 2,190 vs. 1,090,000; P < 0.001), and were independently associated with treatment outcomes (P = 0.003). A threshold of 250 I(eot) was shown to identify sustained virological response (SVR) with high positive predictive value (93%) and good diagnostic accuracy (81%). The time taken to attain 250 I(eot) ranged from 3 to 11 months in patients with hepatitis C virus (HCV) genotypes 2 or 3 and from 3 to 18 months in those with HCV genotypes 1 or 4. Overall, the duration of therapy would have been 49 months less than that suggested by the most recent algorithms based on a rapid virological response (RVR) at week 4.

Furfuryl alcohol polymerization in H-Y confined spaces: reaction mechanism and structure of carbocationic intermediates.

The acid-catalyzed polymerization and resinification, in the 300-673 K interval, of furfuryl alcohol adsorbed in the framework of a protonic Y zeolite is studied by means of FTIR, Raman, and UV-vis spectroscopies. The idea is that restricted spaces can impose a constraint to the growth of the oligomeric chains, therefore moderating the formation of conjugated sequences responsible for the color of the products and allowing their observation by means of spectroscopic techniques. The detailed study of the evolution of UV-vis, FTIR, and Raman spectra upon dosed amount, contact time, and temperature has allowed the spectroscopic features of some of the single species, either neutral and positively charged (carbocationic intermediates), to be singled out and assigned to understand the mechanism of initiation. The vibrational assignments have been confirmed by computer simulations on model compounds and compared with the results of the mechanistic description of the reaction mechanism made in the past (Choura, et al. Macromolecules 1996, 29, 3839-3850). The spectroscopic methods have been applied in a large temperature range in order to follow also the formation of more complex products into the pores, associated with longer conjugated sequences, gradually filling the open spaces of the zeolite. For samples contacted with furfuryl alcohol at 673 K, this methodology gives information also on the incipient carbonization process, leading to the formation of a carbonaceous replica phase inside the internal porosity of the zeolite.

Adsorption properties of HKUST-1 toward hydrogen and other small molecules monitored by IR.

Among microporous systems metal organic frameworks are considered promising materials for molecular adsorption. In this contribution infrared spectroscopy is successfully applied to highlight the positive role played by coordinatively unsaturated Cu2+ ions in HKUST-1, acting as specific interaction sites. A properly activated material, obtained after solvent removal, is characterized by a high fraction of coordinatively unsaturated Cu2+ ions acting as preferential adsorption sites that show specific activities towards some of the most common gaseous species (NO, CO2, CO, N2 and H2). From a temperature dependent IR study, it has been estimated that the H2 adsorption energy is as high as 10 kJ mol(-1). A very complex spectral evolution has been observed upon lowering the temperature. A further peculiarity of this material is the fact that it promotes ortho-para conversion of the adsorbed H2 species.

Emergence of hepatitis B virus quasispecies with lower susceptibility to nucleos(t)ide analogues during lamivudine treatment.

OBJECTIVES: We studied the impact of hepatitis B virus (HBV) polymerase/reverse transcriptase (Pol/Rt) heterogeneity on adefovir rescue therapy in 34 consecutive chronic hepatitis B patients with viral breakthrough during lamivudine monotherapy. METHODS: The Pol/Rt A-F domains were directly sequenced in all patients at baseline, and 12 and 24 months. Response to therapy was evaluated at 3, 6, 12 and 24 months by quantitative HBV-DNA. RESULTS: Primary treatment failures did not occur. At 6 months 24/34 (70.6%) patients had viraemia<10(4) copies/mL [initial viral response (IVR)]; at 12 and 24 months 23 (71.9%) and 26 (81.3%) of 32 had HBV-DNA<200 copies/mL [complete viral response (CVR)]. IVR or CVR patients did not show viral breakthroughs, which occurred in one of the six remaining patients. All but three patients had baseline rtM204I/V substitutions associated with rtL180M in 23, rtL80I/V in 14, rtV173L in 4, rtT184S in 3, rtQ215S in 2 and rtA181S in 2 cases. rtA181S without rtM204I/V was found in one patient. Four of the six patients (67%) without 24 month CVR showed rtA181S or rtT184S substitutions either alone or with typical lamivudine resistance profiles. Baseline HBV-DNA levels were negatively associated with IVR (univariate analysis, P=0.023). At least one of rtA181S and rtT184S substitutions correlated negatively with IVR and CVR (univariate analysis, P=0.001) and was independently associated with absence of CVR (P = 0.016). CONCLUSIONS: Lamivudine monotherapy favours the emergence of viral quasispecies that influence the response rate to adefovir rescue therapy independently from baseline viraemia and lower the susceptibility to other nucleos(t)ide analogues.

Transient elastography: a new surrogate marker of liver fibrosis influenced by major changes of transaminases.

Liver stiffness was measured by transient elastography (FibroScan) in 228 consecutive patients with chronic viral hepatitis, with (115) or without cirrhosis (113), to study its correlations with serum transaminases [alanine aminotransferase (ALT)], fibrosis stage and surrogate noninvasive markers of fibrosis (APRI, FORNS, FibroTest and hyaluronic acid). The dynamic profiles of serum transaminases and liver stiffness were compared by multiple testing in 31 patients during a 6-month follow-up. We identified 8.3 and 14 kPa as the fibrosis >/=F2 and cirrhosis cut-offs, respectively: their sensitivities were 85.2%/78.3%; specificities 90.7%/98.2%; positive predictive values 93.9%/97.8%; negative predictive values 78.8%/81.6%; diagnostic accuracies 87.3%/88.2%. FibroScan performed better than the other surrogate markers of fibrosis (P < 0.001). Other than fibrosis, other factors independently associated with liver stiffness were ALT for all patients and chronic hepatitis patients (P < 0.001), and 12-month persistently normal ALT (biochemical remission, P < 0.001) in cirrhotics. In patients with biochemical remission either spontaneous or after antiviral therapy (48 of 228, 21%), liver stiffness was lower than in patients with identical fibrosis stage, but elevated ALT (P < 0.001). The liver stiffness dynamic profiles paralleled those of ALT, increasing 1.3- to 3-fold during ALT flares in 10 patients with hepatitis exacerbations. Liver stiffness remained unchanged in 21 with stable biochemical activity (P = 0.001). In conclusion, transient elastography is a new liver parameter that behaves as a reliable surrogate marker of fibrosis in chronic viral hepatitis patients, provided that its relationship with major changes of biochemical activity is taken into account.

Predicting response to peginterferon alpha-2a, lamivudine and the two combined for HBeAg-negative chronic hepatitis B.

OBJECTIVE: In a trial of patients with hepatitis B e antigen (HBeAg)-negative chronic hepatitis B, 24 week post-treatment biochemical and virological response rates with peginterferon alpha-2a with or without lamivudine were significantly higher than with lamivudine alone. The effect of pre-treatment factors on post-treatment responses was investigated. METHODS: Multivariate analyses were performed using available data from 518 patients treated with peginterferon alpha-2a with or without lamivudine, or with lamivudine alone. A post-treatment response was defined as alanine aminotransferase (ALT) normalisation and hepatitis B virus (HBV) DNA level of <20,000 copies/ml. RESULTS: In logistic regression analyses across all treatment arms, peginterferon alpha-2a (with or without lamivudine) therapy, younger age, female gender, high baseline ALT, low baseline HBV DNA and HBV genotype were identified as significant predictors of combined response at 24 weeks post-treatment. In the peginterferon alpha-2a and lamivudine monotherapy arms, patients with genotypes B or C had a higher chance of response than genotype D infected patients (p<0.001), the latter responding better to the combination than to peginterferon alpha-2a monotherapy (p = 0.015). At 1 year post-treatment, response rates by intention-to-treat analysis were 19.2% for the peginterferon alpha-2a, 19.0% for the combination, and 10.0% for the lamivudine groups, with genotypes B or C associated with a sustained combined response to peginterferon alpha-2a with or without lamivudine therapy. CONCLUSIONS: Baseline ALT and HBV DNA levels, patient age, gender, and infecting HBV genotype significantly influenced combined response at 24 weeks post-treatment, in patients treated with peginterferon alpha-2a and/or lamivudine. At 1 year post-treatment HBV genotype was significantly predictive of efficacy for patients treated with peginterferon alpha-2a with or without lamivudine.

Animal model for liver cell banking from non-heart beating donors after prolonged ischaemia time.

BACKGROUND AND AIM: Although there is a growing interest on the use of non-heart beating donors to enlarge the liver donor pool, livers with prolonged warm ischaemia time are not currently considered for organ transplantation. We hypothesised that these organs may represent a source of hepatocytes for cell transplantation and/or use in bioartificial liver devices. Thus, we investigated if prolonged ischaemia could influence the recovery and viability of functional hepatocytes dissociated from rat livers. METHODS: Hepatocytes were isolated from the liver within 15 min after death (t=15 min) and after 4, 8 and 12h of ischaemia. Cells were either maintained in culture or cryopreserved. In all products, we evaluated cell recovery and viability, hepatocyte markers and cellular functions, including albumin and urea production. RESULTS: The number of cells per gram of tissue was similar at 15 min, 4 and 8h, while it was significantly decreased at 12h. About 0.2 x 10(6) viable cells expressing hepatocyte markers and producing albumin and urea were isolated up to 8h of ischaemia per gram of tissue. CONCLUSIONS: Recovery of viable and functional hepatocytes seems possible after prolonged ischaemia time. These data warrant the evaluation of hepatocyte isolation from human livers of non-heart beating donors.

Validation and comparison of different PCR-based methods for detection of hepatitis B virus precore region mutants.

Hepatitis virus variants detection is useful in clinical practice; however, methods that are used for their identification may influence the results significantly. Three PCR-based assays for quantitation of G1896A precore HBV mutants: two allele specific PCRs, single tube (single-AS-PCR) with enzymatic restriction or separate tubes (twin-AS-PCR) and one oligohybridization assay (OA) with three probes were developed and standardized. Wild type and mutant plasmids and 10 sera were used as reference. All methods had sensitivity limits of 10(4)copies/ml and their specificity encompassed 3 logs (10(4)-10(7)copies/ml) with dynamic ranges of logs for OA, twin-AS-PCR and single-AS-PCR, respectively. Single-AS-PCR and OA detected minor viral populations when their relative prevalence was at least 10% of the overall viral population whereas their detection by twin-AS-PCR ranged from 0.1 to 10% for samples with 10(7) and 10(5)copies/ml viral loads, respectively. Twin-AS-PCR was the most sensitive to detect the minor viral population, whereas single-AS-PCR and OA were more accurate to quantify the relative proportions of the two viral populations independently of the overall viral load. In conclusion, an accurate characterization of HBV precore heterogeneity should be warranted by a careful choice of the most appropriate assay according to the aim of the study.

Electronic and vibrational properties of a MOF-5 metal-organic framework: ZnO quantum dot behaviour.

UV-Vis DRS and photoluminescence (PL) spectroscopy, combined with excitation selective Raman spectroscopy, allow us to understand the main optical and vibrational properties of a metal-organic MOF-5 framework. A O(2-)Zn(2+)[rightward arrow] O(-)Zn(+) ligand to metal charge transfer transition (LMCT) at 350 nm, testifies that the Zn(4)O(13) cluster behaves as a ZnO quantum dot (QD). The organic part acts as a photon antenna able to efficiently transfer the energy to the inorganic ZnO-like QD part, where an intense emission at 525 nm occurs.

Treatment of chronic hepatitis B: from research to clinical practice via the consensus conferences.

The aim of antiviral therapy of chronic hepatitis B is to control Hepatitis B Virus (HBV) replication and to cure liver disease avoiding the progression of chronic hepatitis to cirrhosis and the end stage complications of cirrhosis. HBeAg/anti-HBe seroconversion is the hallmark of response in hepatitis B "e" antigen (HBeAg) positive patients. In the patients with antibody against HBeAg (anti-HBe positive) the combination of HBV DNA and anti-HBc IgM tests provides adequate diagnostic accuracy. Patients with biochemical and/or histological disease activity are eligible to therapy. The drug choice is based on age, disease severity, risk of complications, side effects and compliance, particularly in anti-HBe positive patients where prolonged treatment is needed. Interferon (5-6 MU daily or 9-10 MU thrice weekly for 4-6 months) is the first line therapy for HBeAg positive patients and (5-6 MU thrice weekly for 12-24 months) for anti-HBe positive patients. When IFN is contraindicated or ineffective, Lamivudine (100 mg) or Adefovir Dipivoxil (10 mg) are given as long as 4-6 months after HBeAg/anti-HBe seroconversion or for long-term treatments in HBeAg positive non-responders and anti-HBe positive patients. Patients with more advanced forms of cirrhosis and portal hypertension are to be treated within liver transplantation programs. Fifteen to 30% of treated patients achieve sustained response and more than 60% of them experience long-term disease remission during therapy. In perspectives, currently available molecular and immunologic tools and modelling of viral dynamics will help to address the therapy issue with more complex, efficacious and individually tailored treatment schedules.