Immunohistochemical and radiological characterization of wound healing in porcine liver after radiofrequency ablation

Background: Radiofrequency ablation (RFA) is a minimal invasive therapeutic option for patients with hepatocellular carcinoma or liver metastases. We investigated RFA-induced cellular changes in the liver of pigs. Material and Methods: Healthy pigs (n=18) were sacrificed between day 0 and 3 months after RFA. The wound healing process was evaluated by computed tomography (CT), chromotrope anilinblue (CAB) staining of large-scale and standard tissue sections. Immunohistochemistry (IHC) for heat shock protein 70, Caspase-3, Ki67, Reelin, Vinculin, Vimentin and αSMA was perfomed. Results: One day after RFA, CAB staining showed cell damage and massive hyperaemia. All IHC markers were predominantly expressed at the outer borders of the lesion, except Reelin, which was mainly detected in untreated liver regions. By staining for Hsp70, the heat stress during RFA was monitored, which was most distinct 1-2 days after RFA. CT revealed decreased lesion size after one week. Development of a Vimentin and α-SMA positive fibrotic capsule was observed. Conclusion: In the early phase signs of cell damage, apoptosis and proliferation are dominant. Reduced expression of Reelin suggests a minor role of hepatic stellate cells in the RFA zone. After one week myofibroblasts become prominent and contribute to the development of the fibrotic capsule. This elucidates the pathophysiology of RFA and could contribute to the future optimization of RFA procedures

Triple Therapy with First Generation Protease Inhibitors for Hepatitis C Markedly Impairs Function of Neutrophil Granulocytes.

First-generation HCV protease inhibitors represent a milestone in antiviral therapy for chronic hepatitis C infection (CHC), but substantially increased rates of viral clearance are offset by increased rates of infection and infection-associated deaths, especially of patients with advanced liver disease. We aimed to assess whether first generation protease inhibitors interfere with neutrophil function. We included 108 consecutive, retrospective CHC patients and 44 consecutive, prospective CHC patients who were treated with peginterferon and ribavirin with or without protease inhibitors according to the guidelines in the period of November 2012 to June 2015. 33 healthy volunteers served as controls. Infection data were evaluated in all patients. Neutrophil phagocytosis, oxidative burst, elastase and diamine oxidase levels during 12 weeks of triple (n = 23) or dual therapy (n = 21) were studied in the prospective part. In the retro- and prospective cohorts patients experiencing clinically relevant infections were significantly more frequent during protease inhibitor therapy (31% and 26%) than during therapy with peginterferon and ribavirin (13% and 0%). Neutrophil phagocytosis decreased to 40% of baseline with addition of protease inhibitors to P/R but recovered 6 months after end of treatment. Protease inhibitors also seemed to reduce serum elastase levels but did not impact on gut permeability. Impaired neutrophil function during triple therapy with first generation HCV protease inhibitors may explain the high infection rate associated to these treatments and be of relevance for treatment success and patient survival

Lactobacillus casei Shirota Supplementation Does Not Restore Gut Microbiota Composition and Gut Barrier in Metabolic Syndrome: A Randomized Pilot Study.

Metabolic syndrome is associated with disturbances in gut microbiota composition. We aimed to investigate the effect of Lactobacillus casei Shirota (LcS) on gut microbiota composition, gut barrier integrity, intestinal inflammation and serum bile acid profile in metabolic syndrome. In a single-centre, prospective, randomised controlled pilot study, 28 subjects with metabolic syndrome received either LcS for 12 weeks (n = 13) or no LcS (n = 15). Data were compared to healthy controls (n = 16). Gut microbiota composition was characterised from stool using 454 pyrosequencing of 16S rRNA genes. Serum bile acids were quantified by tandem mass spectrometry. Zonulin and calprotectin were measured in serum and stool by ELISA. Bacteroidetes/Firmicutes ratio was significantly higher in healthy controls compared to metabolic syndrome but was not influenced by LcS. LcS supplementation led to enrichment of Parabacteroides. Zonulin and calprotectin were increased in metabolic syndrome stool samples but not influenced by LcS supplementation. Serum bile acids were similar to controls and not influenced by LcS supplementation. Metabolic syndrome is associated with a higher Bacteroidetes/Firmicutes ratio and gut barrier dysfunction but LcS was not able to change this. LcS administration was associated with subtle microbiota changes at genus level.ClinicalTrials.gov NCT01182844

Characteristics of neutrophils in patient groups and healthy controls.

<p>Phagocytic capacity of neutrophils (a), proportion of inactive neutrophils (b) and PMN elastase levels per 10⁶neutrophils (c) of patients and controls. Ctrl: healthy controls; TPV: telaprevir group; BOC: boceprevir Group; P/R: Dual therapy with peginterferon/ribavirin; B: baseline; 4w: 4 weeks of therapy; 12w: 12 weeks of therapy; F: follow up a: p = 0.021 vs Ctrl; b: p = 0.010 vs Ctrl.</p

Flow diagram of the study progress.

<p>Flow diagram of the study progress.</p

Patient and infection characteristics for 108 retrospectively analysed patients during hepatitis C therapy.

<p>Patient and infection characteristics for 108 retrospectively analysed patients during hepatitis C therapy.</p

Evenness and diversity of gut microbiota in MetS and healthy controls.

<p>MetS: metabolic syndrome.</p><p>Data are given as median (quartiles).</p

Flow diagram of the study progress.

<p>Flow diagram of the study progress.</p

Patient characteristics.

<p>*p<0.05</p><p>**p<0.01</p><p>***p<0.001 compared to the other groups at baseline</p><p>n.a. not available; EOS: end of study; base: baseline; healthy: healthy controls.</p><p>Data are given as mean±SD.</p

Gut microbiota composition in MetS patients and controls.

<p><i>Bacteroidetes</i>/<i>Firmicutes</i> ratio (a) PCoA Plot (Weighted UniFrac, b) and abundance of <i>Parabacteroides</i> (c) concerning <i>Lc</i>S supplementation.</p