The effect of autologous macrophage therapy in cirrhosis in response to individual immune reparative pathways: developing a novel therapy

BACKGROUND: Liver cirrhosis is the end stage of any injury process to the liver. Once established it inevitably progresses to complications such as portal hypertension, cancer and death. There is not cure for liver cirrhosis besides liver transplant. We face an unmet demand for treatment of this condition. The role of macrophages in fibrosis development and resolution in the liver has been extensively investigated. Prof Forbes group invested in the development of autologous macrophage product to promote fibrosis resolution hence cirrhosis regression. This has demonstrated its efficacy and safety in animal models. From these encouraging pre-clinic data a phase 1 first in human clinical trial of autologous activated macrophage product for cirrhotic patients was developed. METHODS: Using an established 3+3 dose escalation model we enrolled a total of 9 subject in the phase 1 trial reaching a maximum achieved and safe dose of 1x10^9 macrophages. In addition to adverse events, dose toxicity and macrophage activation syndrome (MAS) parameter, we evaluated a varied range of circulating cytokines and chemokine pre and post treatment using a commercial kit. Moreover we developed a protocol for P13- magnetic resonance spectrometry (MRS) for the analysis of the metabolically active liver parenchyma. Data from the phase 1 trial were used to improve the autologous cellular produce and phase 2 randomised controlled trial. RESULTS: The autologous activated macrophage produce is demonstrated not to cause any toxicity in this first in human study of cirrhotic population of different aetiology. Cytokine and chemokine analysis supports these findings and specifically demonstrates low levels of IL-8, which represent cardinal feature of MAS. Other interesting cytokine signals may support extra cellular matrix remodelling effect of the autologous macrophage product infusion. In addition we demonstrated a reproducible protocol for MRS in liver disease. DISCUSSION: Autologous activated macrophage infusion did not result in any toxicity in cirrhotic subjects taking part in this study and shows preliminary signs of efficacy in fibrosis resolution both clinically and biochemically. This work places the basis of development of cellular products for treatment of cirrhosis and fibrosis and provides invaluable insight in immune response to cellular treatment

Characterizing the efficacy of a film-forming antitranspirant on raspberry foliar and fruit transpiration

The film-forming antitranspirant, di-1-p-menthene, is able to reduce transpiration in a number of crops, potentially resulting in water savings and improved productivity. The success of the response is, however, dependent on genotype and environmental factors. We aimed to assess the efficacy of this natural terpene polymer on red raspberry (Rubus idaeus, L.) cv. Tulameen leaf water-use efficiency across a 25–40◦ C temperature range under controlled conditions. The film reduced transpiration (E) and was most effective when applied to the lower leaf surface. Leaf net assimilation (A) and stomatal conductance (g) were also curtailed after the application of di-1-p-menthene, and as a consequence intrinsic transpiration efficiency (A/g) and instantaneous transpiration efficiency (ratio of net carbon fixation to water loss, A/E) did not improve. At 40◦ C, gas exchange of both treated and untreated leaves was minimal due to stomatal closure. The antitranspirant was effective at reducing water loss from berries, but only at the immature stages when transpiration rates were naturally high. Further studies are required to determine if the antitranspirant, di-1-p-menthene, will offer protection against dehydration across a range of temperatures and if productivity and berry composition will benefit

Potassium and magnesium mediate the light and co2 photosynthetic responses of grapevines

Potassium (K) and magnesium (Mg) deficiency are common stresses that can impact on grape yield and quality, but their effects on photosynthesis have received little attention. Understanding the diffusional and biochemical limitations to photosynthetic constraints will help to guide improvements in cultural practices. Accordingly, the photosynthetic response of Vitis vinifera cvs. Shiraz and Chardonnay to K or Mg deficiency was assessed under hydroponic conditions using miniature low-nutrient-reserve vines. Photosynthesis was at least partly reduced by a decline in stomatal conductance. Light and CO2-saturated photosynthesis, maximum rate of ribulose 1.5 bisphospate (RuBP) carboxylation (Vcmax) and maximum rate of electron transport (Jmax) all decreased under K and Mg deficiency. Likewise, chlorophyll fluorescence and electron transport were lower under both nutrient deficiencies while dark respiration increased. K deficiency drastically reduced shoot biomass in both cultivars, while root biomass was greatly reduced under both Mg and K deficiency. Taken together, these results indicate that the decrease in biomass was likely due to both stomatal and biochemical limitations in photosynthesis. Optimising photosynthesis through adequate nutrition will thus support increases in biomass with carry-on positive effects on crop yields

The eye as a non-invasive window to the microcirculation in liver cirrhosis: a prospective pilot study

Microcirculatory dysfunction is associated with organ failure, poor response to vasoactive drugs and increased mortality in cirrhosis, but monitoring techniques are not established. We hypothesized that the chorioretinal structures of the eye could be visualized as a non-invasive proxy of the systemic microvasculature in cirrhosis and would correlate with renal dysfunction. Optical Coherence Tomography (OCT) was performed to image the retina in n = 55 cirrhosis patients being assessed for liver transplantation. OCT parameters were compared with established cohorts of age- and sex-matched healthy volunteers (HV) and patients with chronic kidney disease (CKD). Retinal thickness, macular volume and choroidal thickness were significantly reduced relative to HV and comparable to CKD patients (macular volume: HV vs. cirrhosis mean difference 0.44 mm3 (95% CI 0.26–0.61), p ≤ 0.0001). Reduced retinal thickness and macular volume correlated with renal dysfunction in cirrhosis (macular volume vs. MDRD-6 eGFR r = 0.40, p = 0.006). Retinal changes had resolved substantially 6 weeks following transplantation. There was an inverse association between choroidal thickness and circulating markers of endothelial dysfunction (endothelin-1 r = −0.49, p ≤ 0.001; von Willebrand factor r = −0.32, p ≤ 0.05). Retinal OCT may represent a non-invasive window to the microcirculation in cirrhosis and a dynamic measure of renal and endothelial dysfunction. Validation in different cirrhosis populations is now required

Role of Acute Graft-Versus-Host Disease in the Risk of Bacteremia and Invasive Fungal Disease after Allogeneic Hemopoietic Stem Cell Transplantation in Children. Results from a Single-Center Observational Study

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Blood baseline values in female alpine and nera di verzasca goats reared in italy.

The Italian goat autochthonous breeds are appreciated for their milk and characteristics, especially for the rusticity, frugality, fertility and longevity. For these reasons the local goat breeds play an important role in the livestock sector, and it is important to guarantee sanitary strategies control, prevention or treatment of diseases. It is well known that the hematological parameters in goats undergo changes in relation with many factors like breed, age (Piccione et al. 2014), physiological/reproductive status environmental factors and stress (Waziri M.A. et al. 2010).  Based on these differences it is necessary to establish appropriate physiological baseline values for every single breed which could be used in the realistic evaluation physiological or pathological status of the animal (Arfuso et al. 2016). The aim of this work was to evaluate the differences between a local goat breed (Verzasca) and a cosmopolite one (Alpine) from the hematological point of view, and to establish hematological reference values. A total number of 71 female goats, of Alpine (n=37), and Verzasca (n=34) were enrolled for this study, for a total of 716 blood samples. Data were processed by a mixed model-repeated measures ANCOVA in order to evaluate the effects of breed, parity, and season, while baseline values for each breed have been calculated by evaluating the 2.5-97.5th  percentile of variables distribution.The results showed that the breeds differ in a significant manner (Table 1). Verzasca goat shows significantly higher values in the erythroid parameters, whereas the Alpine goat shows higher mean values of leucocyte count and absolute neutrophil count. A further interesting result is the neutrophil lymphocyte ratio which is 0.96  in the Alpine and 0,57 in the Verzasca.The results here presented can add some knowledge to the definition of the health status of the two breeds, evidencing some environmental and physiological variation mechanisms.

Novel sample-substrates for the determination of new psychoactive substances in oral fluid by desorption electrospray ionization-high resolution mass spectrometry

A reliable screening and non invasive method based on the use of microextraction by packed sorbent coupled with desorption electrospray ionization-high resolution mass spectrometry was developed and validated for the detection of new psychoactive substances in oral fluid. The role of different sample substrates in enhancing signal intensity and stability was evaluated by testing the performances of two polylactide-based materials, i.e. non-functionalized and functionalized with carbon nanoparticles, and a silica-based material compared to commercially available polytetrafluorethylene supports. The best results were achieved by using the nonfunctionalized polylactide substrates to efficiently ionize compounds in positive ionization mode, whereas the silica coating proved to be the best choice for operating in negative ionization mode. LLOQs in the low μg/L, a good precision with CV% always lower than 16% and RR% in the 83(±4)-120(±2)% range, proved the suitability of the developed method for the determination of the analytes in oral fluid. Finally, the method was applied for screening oral fluid samples for the presence of psychoactive substances during private parties, revealing mephedrone in only one sample out of 40 submitted to analysis

A Fe2+-dependent self-inhibited state influences the druggability of human collagen lysyl hydroxylase (LH/PLOD) enzymes

Multifunctional human collagen lysyl hydroxylase (LH/PLOD) enzymes catalyze post-translational hydroxylation and subsequent glycosylation of collagens, enabling their maturation and supramolecular organization in the extracellular matrix (ECM). Recently, the overexpression of LH/PLODs in the tumor microenvironment results in abnormal accumulation of these collagen post-translational modifications, which has been correlated with increased metastatic progression of a wide variety of solid tumors. These observations make LH/PLODs excellent candidates for prospective treatment of aggressive cancers. The recent years have witnessed significant research efforts to facilitate drug discovery on LH/PLODs, including molecular structure characterizations and development of reliable high-throughput enzymatic assays. Using a combination of biochemistry and in silico studies, we characterized the dual role of Fe2+ as simultaneous cofactor and inhibitor of lysyl hydroxylase activity and studied the effect of a promiscuous Fe2+ chelating agent, 2,2'-bipyridil, broadly considered a lysyl hydroxylase inhibitor. We found that at low concentrations, 2,2'-bipyridil unexpectedly enhances the LH enzymatic activity by reducing the inhibitory effect of excess Fe2+. Together, our results show a fine balance between Fe2+-dependent enzymatic activity and Fe2+-induced self-inhibited states, highlighting exquisite differences between LH/PLODs and related Fe2+, 2-oxoglutarate dioxygenases and suggesting that conventional structure-based approaches may not be suited for successful inhibitor development. These insights address outstanding questions regarding druggability of LH/PLOD lysyl hydroxylase catalytic site and provide a solid ground for upcoming drug discovery and screening campaigns