Introduction: Tribute to Henry Manne's contributions to law-and-economics

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An exogenous protease increases enzymic activities, microbial numbers and fiber degraation by mixed ruminal microorganisms in continuous culture

Contributed to: Conference on Gastrointestinal Function (Chicago, Illinois, Mar 10-12, 2003).The effects of pH and addition of a protease mixture on the fermentation characteristics of a total mixed ration (TMR) were investigated in a dual-flow continuous culture apparatus, using a 4 x 4 Latin Square design with a 2 x 2 factorial arrangement of treatments. The diet (DM basis) consisted of 30% alfalfa hay, 30% corn silage and 40% rolled corn. The silage and the grain were milled fresh, mixed with the alfalfa and treated with the enzyme (1.5 μL/g feed) daily. Ruminal fluid was collected 2 h post-feeding from 3 lactating dairy cows fed a TMR. Fermenters were fed 80 g DM/d in equal portions every 12 h. Treatments were control (C) and enzymetreated (T) TMR at either high pH (HC and HT) or low pH (LC and LT). The pH was altered by diluting the artificial saliva to 60% of its original composition. Enzymic activities and total and cellulolytic bacterial numbers were determined on the liquid phase of the fermenter contents, 6 h post-feeding. Fiber degradation was determined from the outflow residues. Enzyme addition increased (P < 0.05) xylanase, xylosidase, endoglucanase, and protease activities (608 vs. 750; 0.48 vs. 0.80; 82 vs. 112; and 1.2 vs. 7.5 units for C and T, respectively), whereas it tended (P < 0.12) to increase exoglucanase and glucosidase activities (0.8 vs. 1.4; and 4.7 vs. 5.9 units). However, enzyme did not affect (P = 0.18) arabinofuranosidase activity (5.2 vs. 6.8). Total microbial numbers (expressed as Log10) were increased (P < 0.05) at low pH (9.13 vs. 9.36) but enzyme had no effect (P = 0.13). Cellulolytic bacteria were reduced (P < 0.02) at low pH (3.91 vs. 2.79), with no effect (P = 0.88) of enzyme. Low pH reduced (P < 0.001) NDF, ADF, and cellulose degradation. NDF degradation was increased (P < 0.01) by enzyme addition (20% vs. 27%) but ADF was unaffected (P < 0.20), resulting in an increase (P < 0.001) in hemicellulose degradation. It is speculated that the enzyme removed structural barriers present in the feed, allowing a more rapid colonization of the fiber by ruminal microorganisms.Peer reviewe

Hypoxia up-regulates SERPINB3 through HIF-2\u3b1 in human liver cancer cells.

SERPINB3 is a cysteine-proteases inhibitor up-regulated in a significant number of cirrhotic patients carrying hepatocellular carcinoma (HCC) and recently proposed as a prognostic marker for HCC early recurrence. SERPINB3 has been reported to stimulate proliferation, inhibit apoptosis and, similar to what reported for hypoxia, to trigger epithelial-to-mesenchymal transition (EMT) and increased invasiveness in liver cancer cells. This study has investigated whether SERPINB3 expression is regulated by hypoxia-related mechanisms in liver cancer cells. Exposure of HepG2 and Huh7 cells to hypoxia up-regulated SERPINB3 transcription, protein synthesis and release in the extracellular medium. Hypoxia-dependent SERPINB3 up-regulation was selective (no change detected for SERPINB4) and operated through hypoxia inducible factor (HIF)-2\u3b1 (not HIF-1\u3b1) binding to SERPINB3 promoter, as confirmed by chromatin immuno-precipitation assay and silencing experiments employing specific siRNAs. HIF-2\u3b1-mediated SERPINB3 up-regulation under hypoxic conditions required intracellular generation of ROS. Immuno-histochemistry (IHC) and transcript analysis, performed in human HCC specimens, revealed co-localization of the two proteins in liver cancer cells and the existence of a positive correlation between HIF-2\u3b1 and SERPINB3 transcript levels, respectively. Hypoxia, through HIF-2\u3b1-dependent and redox-sensitive mechanisms, up-regulates the transcription, synthesis and release of SERPINB3, a molecule with a high oncogenic potential

Effect of condensed tannins in the methanogenic potential and in vitro digestion efficiency of ryegrass

Los rumiantes son capaces de degradar y fermentar los componentes de la pared celular de forrajeras. En el retículo-rumen, la fermentación microbiana particiona la energía bruta del sustrato en ácidos grasos volátiles, biomasa microbiana (BM) y gases de desecho (e.g. metano, CH4). Estos últimos son eliminados y representan, no solo contaminación ambiental por su efecto invernadero; sino también una pérdida de la energía disponible en el alimento. Numerosas especies vegetales poseen metabolitos secundarios, tales como los taninos condensados (TC), que, aunque tienen propiedades antimicrobianas y astringentes, su uso racional podría generar un mejor aprovechamiento de los forrajes con potenciales efectos mitigadores del CH4 entérico. En este contexto, se evaluó el efecto de cuatro dosis de TC de Quebracho (Schinopsis balansae, 0, 1,5, 3,0 y 4,5 mg TC cada 100 mg de sustrato; correspondientes a los tratamientos Ctrl, TC1, TC2 y TC3) mediante la técnica de producción de gas in vitro (PGiv), en presencia o ausencia de polietilenglicol (inhibidor específico de los TC), para determinar el impacto en la fermentación ruminal del raigrás (Lolium perenne). La digestibilidad disminuyó en TC3, mientras que la PGiv aumentó asociada a las mayores tasas iniciales. No hubo efecto de los TC sobre la producción de CH4, tampoco en la síntesis de BM. Por otro lado, el agregado de TC disminuyó la concentración de ácido propiónico, aumentado la relación acético: propiónico. Se concluye que los TC de Quebracho a las dosis estudiadas, no produjeron un efecto mitigador del CH4, además de modificar los parámetros de fermentación, sobre todo con una inclusión de 4,5 mg de TC/100 mg de sustrato incubado.Ruminants are capable of degrading and fermenting the forage cell wall components. In reticulo-rumen, the microbial fermentation partition gross energy of the substrate into volatile fatty acids, microbial biomass (MB) and waste gases (e.g. methane, CH4). These gases are eliminated and they represent, not only environmental pollution due to its greenhouse effect, but they also represent loss of the energy available in the forage. Numerous plant species have secondary compounds, such as condensed tannins (CT), although they have antimicrobial and astringent properties, their rational use could generate a better use of forages with potential mitigating effects of enteric CH4. In this context, it was evaluated the effect of four doses of Quebracho (Schinopsis balansae, 0, 1.5, 3.0 and 4.5 mg of CT per 100 mg of substrate, correspond to treatments Ctrl, TC1, TC2 and TC3) using the in vitro gas production technique (ivGP), in the presence or absence of polyethylene glycol (specific inhibitor of CT), to determine the impact on ruminal fermentation of ryegrass (Lolium perenne). The digestibility decreased in TC3, while the ivGP increased asociated with the higher initial rates. There was no effect of the CT on the CH4 production, neither in the synthesis of MB. On the other hand, the addition of CT decreased propionic acid concentration, increasing the acetic: propionic ratio. It is concluded that the CT of Quebracho, at the doses studied, did not produce a CH4 mitigating effect, in addition to modifying the fermentation parameters, especially with an inclusion of 4.5 mg of CT/ 100 mg of incubated substrate.Fil: Cantet, Juan Manuel. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Neumann Reiter, A. M.. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Animal; ArgentinaFil: Colombatto, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Producción Animal. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Unidad Ejecutora de Investigaciones en Producción Animal; ArgentinaFil: Wawrzkiewicz, Marisa. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Animal; ArgentinaFil: Jaurena, Gustavo. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Animal; Argentin

Minor-but-Complex Liver Resection: An Alternative to Major Resections for Colorectal Liver Metastases Involving the Hepato-Caval Confluence

Major hepatectomy (MH) is often considered the only possible approach for colorectal liver metastasis (CRLM) at the hepato-caval confluence (CC), but it is associated with high morbidity and mortality. With the aim to reduce MH, we developed the "minor-but-complex" (MbC) technique, which consists in the resection of less than 3 adjacent liver segments with exposure of the CC and preservation of hepatic outflow until spontaneous maturation of peripheral intrahepatic shunts between main hepatic veins. We have evaluated applicability and outcome of MbC resections for the treatment of CRLM involving the CC. In this retrospective cohort study, all consecutive liver resections (LR) performed for CRLM located in segments 1, 7, 8, or 4a were classified as MINOR - removal of bC - removal of = 3 adjacent segments. The rate of avoided MH was obtained by the difference between the rate of potentially MH (PMH) plus potentially inoperable cases and the rate of the MH performed. Taking into account that postoperative mortality is mainly related to the amount of resected liver, MbC was compared with minor resections for safety, complexity, and outcome. Of the 59 LR analyzed, 29 (49.1%) were deemed PMH and 4 (6.8%) potentially inoperable. Eventually, MH was performed only in 8 (13.5%) with a decrease rate of 42.4%. Minor LR was performed in 23 (39.0%) and MbC LR in 28 (47.5%) patients. Among MbC cases, 32.1% had previous liver treatments, 39.3% required vascular reconstruction (no reconstructed vessel thrombosis occurred before maturation of peripheral intrahepatic shunts between main hepatic veins), and 7.1% had grade IIIb-IV complications, their median hospital stay was 9 days and 90-day mortality was 0%. After a median follow-up of 22.2 months, oncological results were comparable with those of minor resections. MbC hepatectomy lowers the need for MH and allows for the resection of potentially inoperable patients without negative impact on safety and survival

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

Viral Kinetics Suggests a Reconciliation of the Disparate Observations of the Modulation of Claudin-1 Expression on Cells Exposed to Hepatitis C Virus

The tight junction protein claudin-1 (CLDN1) is necessary for hepatitis C virus (HCV) entry into target cells. Recent studies have made disparate observations of the modulation of the expression of CLDN1 on cells following infection by HCV. In one study, the mean CLDN1 expression on cells exposed to HCV declined, whereas in another study HCV infected cells showed increased CLDN1 expression compared to uninfected cells. Consequently, the role of HCV in modulating CLDN1 expression, and hence the frequency of cellular superinfection, remains unclear. Here, we present a possible reconciliation of these disparate observations. We hypothesized that viral kinetics and not necessarily HCV-induced receptor modulation underlies these disparate observations. To test this hypothesis, we constructed a mathematical model of viral kinetics in vitro that mimicked the above experiments. Model predictions provided good fits to the observed evolution of the distribution of CLDN1 expression on cells following exposure to HCV. Cells with higher CLDN1 expression were preferentially infected and outgrown by cells with lower CLDN1 expression, resulting in a decline of the mean CLDN1 expression with time. At the same time, because the susceptibility of cells to infection increased with CLDN1 expression, infected cells tended to have higher CLDN1 expression on average than uninfected cells. Our study thus presents an explanation of the disparate observations of CLDN1 expression following HCV infection and points to the importance of considering viral kinetics in future studies of receptor expression on cells exposed to HCV

Development of Mathematical Models for the Analysis of Hepatitis Delta Virus Viral Dynamics

BACKGROUND: Mathematical models have shown to be extremely helpful in understanding the dynamics of different virus diseases, including hepatitis B. Hepatitis D virus (HDV) is a satellite virus of the hepatitis B virus (HBV). In the liver, production of new HDV virions depends on the presence of HBV. There are two ways in which HDV can occur in an individual: co-infection and super-infection. Co-infection occurs when an individual is simultaneously infected by HBV and HDV, while super-infection occurs in persons with an existing chronic HBV infection. METHODOLOGY/PRINCIPAL FINDINGS: In this work a mathematical model based on differential equations is proposed for the viral dynamics of the hepatitis D virus (HDV) across different scenarios. This model takes into consideration the knowledge of the biology of the virus and its interaction with the host. In this work we will present the results of a simulation study where two scenarios were considered, co-infection and super-infection, together with different antiviral therapies. Although, in general the predicted course of HDV infection is similar to that observed for HBV, we observe a faster increase in the number of HBV infected cells and viral load. In most tested scenarios, the number of HDV infected cells and viral load values remain below corresponding predicted values for HBV. CONCLUSIONS/SIGNIFICANCE: The simulation study shows that, under the most commonly used and generally accepted therapy approaches for HDV infection, such as lamivudine (LMV) or ribavirine, peggylated alpha-interferon (IFN) or a combination of both, LMV monotherapy and combination therapy of LMV and IFN were predicted to more effectively reduce the HBV and HDV viral loads in the case of super-infection scenarios when compared with the co-infection. In contrast, IFN monotherapy was found to reduce the HDV viral load more efficiently in the case of super-infection while the effect on the HBV viral load was more pronounced during co-infection. The results suggest that there is a need for development of high efficacy therapeutic approaches towards the specific inhibition of HDV replication. These approaches may additionally be directed to the reduction of the half-life of infected cells and life-span of newly produced circulating virions