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IntroductionSeveral lines of evidence suggest that the thyroid hormone signaling pathway is altered in patients with NAFLD and that pharmacological strategies to target the thyroid hormone/thyroid hormone nuclear receptor axis (TH/THR) in the liver may exert beneficial effects. In this study, we investigated the effect of TG68, a novel THRβ agonist, on rat hepatic fat accumulation and NAFLD-associated hepatocarcinogenesis.MethodsMale rats given a single dose of diethylnitrosamine (DEN) and fed a high fat diet (HFD) were co-treated with different doses of TG68. Systemic and hepatic metabolic parameters, immunohistochemistry and hepatic gene expression were determined to assess the effect of TG68 on THRβ activation.ResultsIrrespectively of the dose, treatment with TG68 led to a significant reduction in liver weight, hepatic steatosis, circulating triglycerides, cholesterol and blood glucose. Importantly, a short exposure to TG68 caused regression of DEN-induced preneoplastic lesions associated with a differentiation program, as evidenced by a loss of neoplastic markers and reacquisition of markers of differentiated hepatocytes. Finally, while an equimolar dose of the THRβ agonist Resmetirom reduced hepatic fat accumulation, it did not exert any antitumorigenic effect.DiscussionThe use of this novel thyromimetic represents a promising therapeutic strategy for the treatment of NAFLD-associated hepatocarcinogenesis.

BackgroundThyroid hormones (THs) inhibit hepatocellular carcinoma (HCC) through different mechanisms. However, whether microRNAs play a role in the antitumorigenic effect of THs remains unknown.MethodsBy next generation sequencing (NGS) we performed a comprehensive comparative miRNomic and transcriptomic analysis of rat hepatic preneoplastic lesions exposed or not to a short-term treatment with triiodothyronine (T3). The expression of the most deregulated miRs was also investigated in rat HCCs, and in human hepatoma cell lines, treated or not with T3.ResultsAmong miRs down-regulated in preneoplastic nodules following T3, co-expression networks revealed those targeting thyroid hormone receptor-β (Thrβ) and deiodinase1, and Oxidative Phosphorylation. On the other hand, miRs targeting members of the Nrf2 Oxidative Pathway, Glycolysis, Pentose Phosphate Pathway and Proline biosynthesis – all involved in the metabolic reprogramming displayed by preneoplastic lesions– were up-regulated. Notably, while the expression of most miRs deregulated in preneoplastic lesions was not altered in HCC or in hepatoma cells, miR-182, a miR known to target Dio1 and mitochondrial complexes, was down-deregulated by T3 treatment at all stages of hepatocarcinogenesis and in hepatocarcinoma cell lines. In support to the possible critical role of miR-182 in hepatocarcinogenesis, exogenous expression of this miR significantly impaired the inhibitory effect of T3 on the clonogenic growth capacity of human HCC cells.ConclusionsThis work identified several miRNAs, so far never associated to T3. In addition, the precise definition of the miRNA-mRNA networks elicited by T3 treatment gained in this study may provide a better understanding of the key regulatory events underlying the inhibitory effect of T3 on HCC development. In this context, T3-induced down-regulation of miR-182 appears as a promising tool.

The immunomodulatory effects of HLA-G expression and its role in cancers, human liver infections and liver transplantation are well documented, but so far, there are only a few reports addressing autoimmune liver diseases, particularly autoimmune hepatitis (AIH).Method and materialsWe analyzed the genetic and phenotypic characteristics of HLA-G in 205 type 1 AIH patients (AIH-1) and a population of 210 healthy controls from Sardinia (Italy).ResultsAnalysis of the HLA-G locus showed no substantial differences in allele frequencies between patients and the healthy control population. The HLA-G UTR-1 haplotype was the most prevalent in both AIH-1 patients and controls (40.24% and 34.29%). Strong linkage was found between the HLA-G UTR-1 haplotype and HLA-DRB1*03:01 in AIH-1 patients but not controls (D’ = 0.92 vs D’ = 0.50 respectively; P = 1.3x10-8). Soluble HLA-G (sHLA-G) levels were significantly lower in AIH-1 patients compared to controls [13.9 (11.6 – 17.4) U/mL vs 21.3 (16.5 – 27.8) U/mL; P = 0.011]. Twenty-four patients with mild or moderate inflammatory involvement, as assessed from liver biopsy, showed much higher sHLA-G levels compared to the 28 patients with severe liver inflammation [33.5 (23.6 – 44.8) U/mL vs 8.8 (6.1 – 14.5) U/mL; P = 0.003]. Finally, immunohistochemistry analysis of 52 liver biopsies from AIH-1 patients did not show expression of HLA-G molecules in the liver parenchyma. However, a percentage of 69.2% (36/52) revealed widespread expression of HLA-G both in the cytoplasm and the membrane of plasma cells labeled with anti-HLA-G monoclonal antibodies.ConclusionThis study highlights the positive immunomodulatory effect of HLA-G molecules on the clinical course of AIH-1 and how this improvement closely correlates with plasma levels of sHLA-G. However, our results open the debate on the ambiguous role of HLA-G molecules expressed by plasma cells, which are pathognomonic features of AIH-1.

Sickle cell disease (SCD) is a widespread genetic disease associated with severe disability and multi-organ damage, resulting in a reduced life expectancy. None of the existing clinical treatments provide a solution for all patients. Gene therapy and fetal haemoglobin (HbF) reactivation through genetic approaches have obtained promising, but early, results in patients. Furthermore, the search for active molecules to increase HbF is still ongoing. The delta-globin gene produces the delta-globin of haemoglobin A2 (HbA2). Although expressed at a low level, HbA2 is fully functional and could be a valid anti-sickling agent in SCD. To evaluate the therapeutic potential of a strategy aimed to over-express the delta-globin gene in vivo, we crossed transgenic mice carrying a single copy of the delta-globin gene, genetically modified to be expressed at a higher level (activated), with a humanised mouse model of SCD. The activated delta-globin gene gives rise to a consistent production of HbA2, effectively improving the SCD phenotype. For the first time in vivo, these results demonstrate the therapeutic potential of delta-globin, which could lead to novel approaches to the cure of SCD.

Ionic liquids (ILs), named also as liquid salts, are compounds that have unique properties and molecular architecture. ILs are used in various industries; however, due to their toxicity, the ILs' recovery from the postreaction solutions is also a very important issue. In this paper, the possibility of 1,3-dialkylimidazolium IL, especially the

Adult hepatocytes are quiescent cells that can be induced to proliferate in response to a reduction in liver mass (liver regeneration) or by agents endowed with mitogenic potency (primary hyperplasia). The latter condition is characterized by a more rapid entry of hepatocytes into the cell cycle, but the mechanisms responsible for the accelerated entry into the S phase are unknown.Next generation sequencing and Illumina microarray were used to profile microRNA and mRNA expression in CD-1 mice livers 1, 3 and 6 h after 2/3 partial hepatectomy (PH) or a single dose of TCPOBOP, a ligand of the constitutive androstane receptor (CAR). Ingenuity pathway and DAVID analyses were performed to identify deregulated pathways. MultiMiR analysis was used to construct microRNA-mRNA networks.Following PH or TCPOBOP we identified 810 and 527 genes, and 102 and 10 miRNAs, respectively, differentially expressed. Only 20 genes and 8 microRNAs were shared by the two conditions. Many miRNAs targeting negative regulators of cell cycle were downregulated early after PH, concomitantly with increased expression of their target genes. On the contrary, negative regulators were not modified after TCPOBOP, but Ccnd1 targeting miRNAs, such as miR-106b-5p, were downregulated.While miRNAs targeting negative regulators of the cell cycle are downregulated after PH, TCPOBOP caused downregulation of miRNAs targeting genes required for cell cycle entry. The enhanced Ccnd1 expression may explain the more rapid entry into the S phase of mouse hepatocytes following TCPOBOP.

Celem pracy było zbadanie dawek poza obszarem terapeutycznym na wybranych głębokościach i odległościach od osi wiązki (CAX). Na potrzeby badania zaprojektowano i skonstruowano fantom wodny o wymiarach 45cm x 35cm x 90cm, w którym umieszczone zostały detektory termoluminescencyjne (TLD 100). Detektory umieszczone zostały na czterech głębokościach : 2cm, 5cm, 8cm, i 15 cm oraz czterech odległościach (dla każdej głębkości) od osi wiązki: 10, 20, 30 i 40 cm. W osi wiązki zadano dawkę 76 Gy, co odpowiadało 7503 jednostkom monitorowym (MU). Dawka w odległości 10 cm od osi wiązki i 2 cm głębokości była bliska dawce na głębokości 15 cm i wynosiła średnio 2,3 Gy. Pomiary dawek pokazały silną zależność dawki w zależności od odległości od osi wiązki. Dla dystansu 40 cm od CAX na głębokości 2 cm, zmierzono dawkę równą 0,27 Gy (SD= 0,025), na 15cm od CAX 0,116 (SD=0,066). W odległości 30 cm od osi wiązki zależność dawki wraz z głębokością była znacząca na głębokości 2 cm – 0.524 Gy (SD=0,251) oraz na głębokości 15 cm – 0.242 Gy (SD=0,564). 20 cm od osi wiązki na 2 cm głębokości wynosiła 0,925 Gy (SD=0,483) i głębokości 15 cm 0,517 (SD=0,160)

Introduction Contemporary radiotherapy uses a number of highly specialized irradiation techniques dedicated to well-defined clinical diagnoses. Among these methods are techniques to irradiate the skin (TSEI), bone marrow (TMI) or the whole body of the patient (TBI). TBI has over the last century been used in the treatment of a variety of conditions, both benign and malignant. However, its importance has increased with the development of knowledge about the impact of ionizing radiation on the human body and the development of clinical dosimetry techniques. At present, however, this method is primarily used in the treatment of hyperplasia. Aim The general aim of the study is to compare dose distributions at selected points of the anthropomorphic phantom under full body radiation conditions for X: 6MV and 15MV radiation. Specific objectives are defined: comparison of percent depth and function of photon emission profiles: 6MV and 15MV measured with radiofrequency hydrophobic films; measurement of doses in selected cross sections of the anthropomorphic phantom. Material and method A number of measuring devices and materials used in daily work by staff of the Medical Physics Department of the Greater Poland Cancer Centre were used to carry out the study part, but Alderson's anthropomorphic phantom and the radiochromic films in the form of point detectors were essential. In addition to each step of the research part, a special measuring system was prepared to reproduce the conditions prevailing during the TBI session as closely as possible. The research was carried out in three stages: Calibration of radiochromic films; PDD and OCR measurement for: 6MV and 15MV photon beam under TBI conditions; Measurement of dose distribution in selected anthropomorphic phantom's cross sections using radiochromic films in the form of point detectors. Results For the lateral field irradiated with 6MV photon beam, the maximum compliance (less than 2%) was obtained for the elbows at the entrance and in the center of the phantom; abdomen for the detector positioned in the center of the phantom, the lungs at the entrance and the arms in the middle, and the neck at the position of the film at the entrance. In the case of the lateral field X 15MV, the highest correspondence occurred for the points: the head and the PC in the position of the film in the center of the phantom and the entrance neck. In the case of AP/PA fields for 6MV energy, the highest compatibility was obtained for the mediastinum in all positions of the film. A small difference was also obtained for the points: head in the middle and at the output of the beam; as well as PC on the output. For AP/PA X 15MV fields, the highest dose compliance not exceeding 1% was obtained for the location of the neck - at the beam entrance, and the lung and mediastinum at the detector position at the center of the phantom. Conclusions On the basis of measurements of dose distribution at selected points of the patient's body for radiation X: 6MV and 15MV in the TBI procedure, the following conclusions can be made: Gafchromic EBT (radiochromic type film) can be successfully used for dosimetric measurements, among others. Due to their properties, such as the ability to cut from the sheet of film spot detectors of any shape and size, flexibility, low sensitivity to daylight, resistance to humidity, etc.; Their main drawback is the high cost of buying films and the long time required to prepare the detectors and then read the measured doses. Because of the low popularity of point-based EBTs in point dosing, further research is needed to improve their response to ionizing radiation. There is a noticeable increase in the difference between the dose calculated and measured as the distance between the position of individual detectors increases from the center point. The difference between the dose measured and planned in any of the cases examined does not exceed 9%. The measurements show that the method used is fast, accurate, and can be successfully used as a validation tool not only for the TBI procedure but also for other methods of cancer radiotherapy.

Background The calculation and measurement on the surface of the skin presents a significant dosimetric problem because of numerous factors which have an influence on the dose distribution in this region. Aim The overall aim of this study was to check the agreement between doses measured with thermoluminescent detectors (TLD) during tomotherapy photon beam irradiation of the skin area of a solid water cylindrical phantom with doses calculated with Hi-Art treatment planning system (TPS). Material and Method The measurements of the dose were made with the use of a solid water cylindrical phantom - Cheese Phantom. Two bolus phantoms were used: 5 mm and 10 mm Six different planning treatments were generated. The doses were measured using TL detectors. Results In the case of a tumor located near the surface of the skin, the mean dose for 0.5 cm bolus was - 1.94 Gy, and for 1 cm bolus - 2.03 Gy. For the tumor located inside the phantom and organ at risk on the same side that TL detectors, for a 0.5 cm bolus, mean dose was 0.658 Gy, and for a 1 cm bolus, 0.62 Gy. Conclusion The analysis of results showed that the relative percentage difference between measured and planned dose in the field of irradiation was less than 10%, while the largest differences were on the board of the field of radiation and outside of the field of irradiation, where the dose was 0.08 Gy to 1 Gy.

Ionic liquids (ILs), named also as liquid salts, are compounds that have unique properties and molecular architecture. ILs are used in various industries; however, due to their toxicity, the ILs’ recovery from the postreaction solutions is also a very important issue. In this paper, the possibility of 1,3-dialkylimidazolium IL, especially the N,N-dibutylimidazolium chloride ([C4C4IM]Cl) recovery by using the electrodialysis (ED) method was investigated. The influence of [C4C4IM]Cl concentration in diluate solution on the ED efficiency was determined. Moreover, the influence of IL on the ion-exchange membranes’ morphology was examined. The recovery of [C4C4IM]Cl, the [C4C4IM]Cl flux across membranes, the [C4C4IM]Cl concentration degree, the energy consumption, and the current efficiency were determined. The results showed that the ED allows for the [C4C4IM]Cl recovery and concentration from dilute solutions. It was found that the [C4C4IM]Cl content in the concentrates after ED was above three times higher than in the initial diluate solutions. It was noted that the ED of solutions containing 5–20 g/L [C4C4IM]Cl allows for ILs recovery in the range of 73.77–92.45% with current efficiency from 68.66% to 92.99%. The [C4C4IM]Cl recovery depended upon the initial [C4C4IM]Cl concentration in the working solution. The highest [C4C4IM]Cl recovery (92.45%) and ED efficiency (92.99%) were obtained when the [C4C4IM]Cl content in the diluate solution was equal 20 g/L. Presented results proved that ED can be an interesting and effective method for the [C4C4IM]Cl recovery from the dilute aqueous solutions.

Background The diversity in the clinical course of COVID-19 has been related to differences in innate and adaptative immune response mechanisms. Natural killer (NK) lymphocytes are critical protagonists of human host defense against viral infections. It would seem that reduced circulating levels of these cells have an impact on COVID-19 progression and severity. Their activity is strongly regulated by killer-cell immuno-globulin-like receptors (KIRs) expressed on the NK cell surface. The present study’s focus was to investigate the impact of KIRs and their HLA Class I ligands on SARS-CoV-2 infection. Methods KIR gene frequencies, KIR haplotypes, KIR ligands and combinations of KIRs and their HLA Class I ligands were investigated in 396 Sardinian patients with SARS-CoV-2 infection. Comparisons were made between 2 groups of patients divided according to disease severity: 240 patients were symptomatic or paucisymptomatic (Group A), 156 hospitalized patients had severe disease (Group S). The immunogenetic characteristics of patients were also compared to a population group of 400 individuals from the same geographical areas. Results Substantial differences were obtained for KIR genes, KIR haplotypes and KIR-HLA ligand combinations when comparing patients of Group S to those of Group A. Patients in Group S had a statistically significant higher frequency of the KIR A/A haplotype compared to patients in Group A [34.6% vs 23.8%, OR = 1.7 (95% CI 1.1–2.6); P = 0.02, Pc = 0.04]. Moreover, the KIR2DS2/HLA C1 combination was poorly represented in the group of patients with severe symptoms compared to those of the asymptomatic-paucisymptomatic group [33.3% vs 50.0%, OR = 0.5 (95% CI 0.3–0.8), P = 0.001, Pc = 0.002]. Multivariate analysis confirmed that, regardless of the sex and age of the patients, the latter genetic variable correlated with a less severe disease course [ORM = 0.4 (95% CI 0.3–0.7), PM = 0.0005, PMC = 0.005]. Conclusions The KIR2DS2/HLA C1 functional unit resulted to have a strong protective effect against the adverse outcomes of COVID-19. Combined to other well known factors such as advanced age, male sex and concomitant autoimmune diseases, this marker could prove to be highly informative of the disease course and thus enable the timely intervention needed to reduce the mortality associated with the severe forms of SARS-CoV-2 infection. However, larger studies in other populations as well as experimental functional studies will be needed to confirm our findings and further pursue the effect of KIR receptors on NK cell immune-mediated response to SARS-Cov-2 infection.

AimSARS-CoV-2 infection is a world-wide public health problem. Several aspects of its pathogenesis and the related clinical consequences still need elucidation. In Italy, Sardinia has had very low numbers of infections. Taking advantage of the low genetic polymorphism in the Sardinian population, we analyzed clinical, genetic and immunogenetic factors, with particular attention to HLA class I and II molecules, to evaluate their influence on susceptibility to SARS-CoV-2 infection and the clinical outcome.Method and MaterialsWe recruited 619 healthy Sardinian controls and 182 SARS-CoV-2 patients. Thirty-nine patients required hospital care and 143 were without symptoms, pauci-symptomatic or with mild disease. For all participants, we collected demographic and clinical data and analyzed the HLA allele and haplotype frequencies.ResultsMale sex and older age were more frequent in hospitalized patients, none of whom had been vaccinated during the previous seasonal flu vaccination campaignes. Compared to the group of asymptomatic or pauci-symptomatic patients, hospitalized patients also had a higher frequency of autoimmune diseases and glucose-6-phosphate-dehydrogenase (G6PDH) deficiency. None of these patients carried the beta-thalassemia trait, a relatively common finding in the Sardinian population. The extended haplotype HLA-A*02:05, B*58:01, C*07:01, DRB1*03:01 [OR 0.1 (95% CI 0–0.6), Pc = 0.015] was absent in all 182 patients, while the HLA-C*04:01 allele and the three-loci haplotype HLA-A*30:02, B*14:02, C*08:02 [OR 3.8 (95% CI 1.8–8.1), Pc = 0.025] were more frequently represented in patients than controls. In a comparison between in-patients and home care patients, the HLA-DRB1*08:01 allele was exclusively present in the hospitalized patients [OR > 2.5 (95% CI 2.7–220.6), Pc = 0.024].ConclusionThe data emerging from our study suggest that the extended haplotype HLA-A*02:05, B*58:01, C*07:01, DRB1*03:01 has a protective effect against SARS-CoV-2 infection in the Sardinian population. Genetic factors that resulted to have a negative influence on the disease course were presence of the HLA-DRB1*08:01 allele and G6PDH deficiency, but not the beta-thalassemic trait. Absence of influenza vaccination could be a predisposing factor for more severe disease.

Activation of the Nrf2-Keap1 pathway, the main intracellular defense against environmental stress, has been observed in several human cancers, including hepatocellular carcinoma (HCC). Here, we assessed whether distinct mechanisms of activation may be involved at different stages of hepatocarcinogenesis. We adopted an experimental model consisting of treatment with diethylnitrosamine (DENA) followed by a choline-devoid methionine-deficient (CMD) diet for 4 months. The CMD diet was then replaced with a basal diet, and the animals were killed at 6, 10 or 13 months after DENA injection. Nrf2 activation occurred at early steps of hepatocarcinogenesis and persisted throughout the tumorigenic process. While Nrf2 mutations were extremely frequent at early steps (90%), their incidence diminished with the progression to malignancy (25%). Conversely, while p62 was almost undetectable in early nodules, its accumulation occurred in HCCs, suggesting that Nrf2 pathway activation at late stages is mainly due to Keap1 sequestration by p62. We demonstrate that, in a model of hepatocarcinogenesis resembling human non-alcoholic fatty liver disease, Nrf2 mutations are the earliest molecular changes responsible for the activation of the Nrf2-Keap1 pathway. The progressive loss of mutations associated with a concomitant p62 accumulation implies that distinct mechanisms are responsible for Nrf2-Keap1 pathway activation at different stages of hepatocarcinogenesis.

Background The AOAC Stakeholder Panel on Strategic Food Analytical Methods approved Standard Method Performance Requirements (SMPR®) 2018.009 for lactose in low-lactose or lactose-free milk, milk products, and products containing dairy ingredients. The LactoSens®R Method is a biosensor assay kit developed for the determination of lactose in a variety of lactose-free or low-lactose milk, dairy, and infant formula products produced with yeast-neutral lactases. Objective In response to a call for methods, the LactoSensR method was validated in a single laboratory study with comparison to SMPR 2018.009. Method The LactoSensR method was evaluated for calibration, interference, repeatability, recovery, and robustness. In a method comparison study samples naturally containing low levels of lactose were evaluated using LactoSensR and an accredited high-performance anion-exchange chromatography with pulsed amperometric detection. Results Calibration with lactose standard solutions was shown to be linear and the method was shown to be free of interference from a variety of sugars, vitamins, alcohols, flavorings, and other compounds. Matrix studies, including 85 spiked materials, 55 products naturally containing lactose, and 13 reference materials, resulted in RSDr of 0–10.5% at 8–100 mg lactose/100 g and 0.2–5.4% at >100 mg lactose/100 g for milk and dairy products and 1.0–6.8% for infant formula, in compliance with SMPR 2018.009 with few exceptions. Recovery was 85.0–110.3% at 8–100 mg lactose/100 g and 85.6–109.7% at >100 mg lactose/100 g for milk and dairy products and 91.1–97.0% for infant formula, also meeting the performance requirements with few exceptions. The method was shown to be robust to changes in ambient temperature, sample temperature, and sample volume. Conclusions The LactoSensR method compares favorably with the requirements of SMPR 2018.009 and should be adopted as a First Action AOAC Official MethodSM. Highlights The LactoSensR method is a fast, easy-to-use method that meets the requirements of SMPR 2018.009.

Activation of Wnt/β-catenin signaling is frequent in human and rodent hepatocarcinogenesis. Although in mice the tumor-promoting activity of agonists of constitutive androstane receptor (CAR) occurs by selection of carcinogen-initiated cells harboring β-catenin mutations, the molecular alterations leading to hepatocellular carcinoma (HCC) development by the CAR agonist 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCP) in the absence of genotoxic injury are unknown. Here, we show that CAR activation per se induced HCC in mice and that 91% of them carried β-catenin point mutations or large in-frame deletions/exon skipping targeting Ctnnb1 exon 3. Point mutations in HCCs induced by TCP alone displayed different nucleotide substitutions compared with those found in HCCs from mice pretreated with diethylnitrosamine. Moreover, unlike those occurring in HCCs from diethylnitrosamine + TCP mice, they did not result in increased expression of β-catenin target genes, such as Glul, Lgr5, Rgn, Lect2, Tbx3, Axin2, and Ccnd1, or nuclear translocation of β-catenin compared with the control liver. Remarkably, in the nontumoral liver tissue, chronic CAR activation led to down-regulation of these genes and to a partial loss of glutamine synthetase-positive hepatocytes. These results show that, although chronic CAR activation per se induces HCCs carrying β-catenin mutations, it concurrently down-regulates the Wnt/β-catenin pathway in nontumoral liver. They also indicate that the relationship between CAR and β-catenin may be profoundly different between normal and neoplastic hepatocytes.

Activation of thyroid hormone receptor β (THRβ) has shown beneficial effects on metabolic alterations, including non-alcoholic fatty liver disease (NAFLD). Here, we investigated the effect of TG68, a novel THRβ agonist, on fatty liver accumulation and liver injury in mice fed a high-fat diet (HFD). C57BL/6 mice fed HFD for 17 or 18 weeks, a time when all mice developed massive steatohepatitis, were then given TG68 at a dose of 9.35 or 2.8 mg/kg for 2 or 3 weeks, respectively. As a reference compound, the same treatment was adopted using equimolar doses of MGL-3196, a selective THRβ agonist currently in clinical phase III. The results showed that treatment with TG68 led to a reduction in liver weight, hepatic steatosis, serum transaminases, and circulating triglycerides. qRT-PCR analyses demonstrated activation of THRβ, as confirmed by increased mRNA levels of Deiodinase-1 and Malic enzyme-1, and changes in lipid metabolism, as revealed by increased expression of Acyl-CoA Oxidase-1 and Carnitine palmitoyltransferase-1. The present results showed that this novel THRβ agonist exerts an anti-steatogenic effect coupled with amelioration of liver injury in the absence of extra-hepatic side effects, suggesting that TG68 may represent a useful tool for the treatment of NAFLD.

Background Although the effects of high dose radiation on human cells and tissues are relatively well defined, there is no consensus regarding the effects of low and very low radiation doses on the organism. Ionizing radiation has been shown to induce gene mutations and chromosome aberrations which are known to be involved in the process of carcinogenesis. The induction of secondary cancers is a challenging long-term side effect in oncologic patients treated with radiation. Medical sources of radiation like intensity modulated radiotherapy used in cancer treatment and computed tomography used in diagnostics, deliver very low doses of radiation to large volumes of healthy tissue, which might contribute to increased cancer rates in long surviving patients and in the general population. Research shows that because of the phenomena characteristic for low dose radiation the risk of cancer induction from exposure of healthy tissues to low dose radiation can be greater than the risk calculated from linear no-threshold model. Epidemiological data collected from radiation workers and atomic bomb survivors confirms that exposure to low dose radiation can contribute to increased cancer risk and also that the risk might correlate with the age at exposure. Conclusions Understanding the molecular mechanisms of response to low dose radiation is crucial for the proper evaluation of risks and benefits that stem from these exposures and should be considered in the radiotherapy treatment planning and in determining the allowed occupational exposures.

To measure out-of-field doses in a phantom model to better quantify this radiation.The individual contribution of photons and neutrons to the total out-of-field dose for 6 MV and 20 MV photons at open beam were measured in a purpose-designed water phantom. Radiation doses were measured at seven separate points (P1-P7) in the phantom with thermoluminescent detectors (TLD 100, 600, and 700) and GAFchromic™ EBT films.At a prescribed dose of 75Gy to the isocentre, the photon dose level in the close-to-field area (P2) ranged from 2.0-2.5Gy for 6 MV and 1.5-2.0Gy for 20 MV; the total out-of-field doses at P2 and P7, respectively, were estimated to be as follows: for 6 MV: TLD 100 (3.23% and0.14%); radiochromic film (2.52% and0.03%); and for 20 MV: TLD 100 (2.94% and0.78%); TLD 700 (2.02% and0.14%); and radiochromic film (1.73% and0.01%). Although the dose decreased rapidly as the distance from the central beam axis increased, even distant doses could be as high as several centigrays. The neutron dose for 20 MV photons at a distance of 25cm from the isocentre was 4.0mSv/Gy.Our results show that in the close-to-field area, the dose level could be as high as 1.5Gy assuming a prescribed dose of 75Gy to the isocentre. By contrast, the doses delivered to more distant areas from the planning target volume were much lower (centigrays). These findings show that both 6 MV and 20 MV photons could produce dosimetrically important dose levels outside of the field. The data reported here may be of value to study the potential impact of even very low doses of radiation on human tissues.

Purpose Patients who undergo external beam radiotherapy are at risk of developing second tumours due to scattered radiation outside the path of the primary beam. The aim of this study was to experimentally determine the in vitro radiobiological effects of scattered radiation in cells located outside the primary photon beam and to compare this to the effects that occur in cells inside the primary beam. The comparison was performed by assessing cell viability, DNA damage, and apoptosis. Material and methods Cells from the human breast cancer line MDA-MB-231 were inserted in a water phantom and irradiated at varying doses (1.5, 2.0, 2.5, and 3.0 Gy). The cells were placed at two geometrical points: in the central beam axis and at 10 cm out-of-field. The dose was constant in both geometrical points. Survival fraction, number of DNA double strand-breaks, and cleaved poly-(ADP-ribose) polymerase (PARP) levels were determined by clonogenic assay and flow cytometry. Results A slight, non-significant decrease of 3 to 5% in cell survival fraction was observed in cells irradiated outside the primary field. The number of PARP-positive cells and DNA double strand-breaks both increased after out-of-field irradiation. Conclusion Scattered irradiation appears to induce an in vitro biological response on out-of-field cells that is stronger than the effect of primary radiation on in-field cells, independent of the bystander effect. These findings suggest that the biological response of healthy tissues outside the primary beam might be higher than previously believed.

Purpose During radiotherapy, leakage from the machine head and collimator expose patients to out-of-field irradiation doses, which may cause secondary cancers. To quantify the risks of secondary cancers due to out-of-field doses, it is first necessary to measure these doses. Since most dosimeters are energy-dependent, it is essential to first determine the type of photon energy spectrum in the out-of-field area. The aim of this study was to determine the mean photon energy values for the out-of-field photon energy spectrum for a 6 MV photon beam using the GEANT 4–Monte Carlo method. Material and methods A specially-designed large water phantom was simulated with a static field at gantry 0°. The source-to-surface distance was 92 cm for an open field size of 10 × 10 cm2. The photon energy spectra were calculated at five unique positions (at depths of 0.5, 1.6, 4, 6, 8, and 10 cm) along the central beam axis and at six different off-axis distances. Results Monte Carlo simulations showed that mean radiation energy levels drop rapidly beyond the edge of the 6 MV photon beam field: at a distance of 10 cm, the mean energy level is close to 0.3 MeV versus 1.5 MeV at the central beam axis. In some cases, the energy level actually increased even as the distance from the field edge increased: at a depth of 1.6 cm and 15 cm off-axis, the mean energy level was 0.205 MeV versus 0.252 MeV at 20 cm off-axis. Conclusion The out-of-field energy spectra and dose distribution data obtained in this study with Monte Carlo methods can be used to calibrate dosimeters to measure out-of-field radiation from 6 MV photons.

The paper presents the research results on the possibility of the integration of advanced oxidation process (AOP) involving Fenton reaction with nanofiltration (NF) for dairy effluent treatment. It has been found that Fenton oxidation reduces organic compounds, total phosphorus and total nitrogen. However, NF enables high ions retention derived from both effluent and Fenton oxidation components. As a result, it was possible to obtain water, which, without any harmful effects, could be discharged into environment. This water also fulfilled most of the requirements to be reused in dairy industrial plant.

Introduction. The rapid development of new radiotherapy technologies, such as intensity modulated radiotherapy (IMRT) or tomotherapy, has resulted in the capacity to deliver a more homogenous dose in the target. However, the higher doses associated with these techniques are a reason for concern because they may increase the dose outside the target. In the present study, we compared 3DCRT, IMRT and tomotherapy to assess the doses to organs at risk (OARs) resulting from photon beam irradiation and scattered neutrons. Material and methods. The doses to OARs outside the target were measured in an anthropomorphic Alderson phantom using thermoluminescence detectors (TLD 100) 6Li (7.5%) and 7Li (92.5%). The neutron fluence rate [cm−2·s−1] at chosen points inside the phantom was measured with gold foils (0.5 cm diameter, mean surface density of 0.108 g/cm3). Results. The doses [Gy] delivered to the OARs for 3DCRT, IMRT and tomotherapy respectively, were as follows: thyroid gland (0.62 ± 0.001 vs. 2.88 ± 0.004 vs. 0.58 ± 0.003); lung (0.99 ± 0.003 vs. 4.78 ± 0.006 vs. 0.67 ± 0.003); bladder (80.61 ± 0.054 vs. 53.75 ± 0.070 vs. 34.71 ± 0.059); and testes (4.38 ± 0.017 vs. 6.48 ± 0.013 vs. 4.39 ± 0.020). The neutron dose from 20 MV X-ray beam accounted for 0.5% of the therapeutic dose prescribed in the PTV. The further from the field edge the higher the contribution of this secondary radiation dose (from 8% to ~45%). Conclusion. For tomotherapy, all OARs outside the therapeutic field are well-spared. In contrast, IMRT achieved better sparing than 3DCRT only in the bladder. The photoneutron dose from the use of high-energy X-ray beam constituted a notable portion (0.5%) of the therapeutic dose prescribed to the PTV.

Objectives Although the hepatomitogenic activity of triiodothyronine (T3) is well established, the wide range of harmful effects exerted by this hormone precludes its use in liver regenerative therapy. Selective agonists of the beta isoform of thyroid hormone receptor (TRβ) do not exhibit T3‐induced cardiotoxicity and show a good safety profile in patients with NASH. The aim of this study was to investigate whether two novel TRβ agonists, the prodrug TG68 and the active compound IS25 could stimulate hepatocyte proliferation without T3/TRα‐dependent side effects. Methods Rats were treated with three different doses (12.5, 25 and 50 μg/100 g body weight) for one week. Hepatocyte proliferation, liver injury and serum biochemical parameters were measured by immunohistochemistry, qRT‐PCR and Western blot. Results Both drugs increased hepatocyte proliferation as assessed by bromodeoxyuridine incorporation (from 14% to 28% vs 5% of controls) and mitotic activity. Enhanced proliferation occurred in the absence of significant signs of liver injury as shown by lack of increased serum transaminase levels or of apoptosis. No cardiac or renal hypertrophy typically associated with treatment with T3 was observed. Importantly, no proliferation of pancreatic acinar cells, such as that seen after administration of T3 or the TRβ agonist GC1 was detected following either TG68 or IS25, demonstrating the hepato‐specificity of these novel TRβ agonists. Conclusions The present study shows that TG68 and IS25 induce massive hepatocyte proliferation without overt toxicity. Hence, these agents may have a significant clinical application for regenerative therapies in liver transplantation or other surgical settings.

Although triiodothyronine (T3) induces several beneficial effects on lipid metabolism, its use is hampered by toxic side-effects, such as tachycardia, arrhythmia, heart failure, bone and muscle catabolism and mood disturbances. Since the α isoform of thyroid hormone receptors (TRs) is the main cause of T3-related harmful effects, several efforts have been made to develop selective agonists of the β isoform that could induce some beneficial effects (i.e. lowering triglyceride and cholesterol levels reducing obesity and improving metabolic syndrome), while overcoming most of the adverse T3-dependent side effects. Herein, we describe the drug discovery process sustained by ADME-Toxicity analysis that led us to identify novel agonists with selectivity for the isoform TRβ and an acceptable off-target and absorption, distribution metabolism, excretion and toxicity (ADME-Tox) profile. Within the small series of compounds synthesized, derivatives 1 and 3, emerge from this analysis as “potentially safe” to be engaged in preclinical studies. In in vitro investigation proved that both compounds were able to reduce lipid accumulation in HepG2 and promote lipolysis with comparable effects to those elicited by T3, used as reference drug. Moreover, a preliminary in vivo study confirmed the apparent lack of toxicity, thus suggesting compounds 1 and 3 as new potential TRβ-selective thyromimetics.

In this work, the use of innovation technique like plasma treatment was introduced in modification of surfaces of polymeric membranes. This creates a great potential in the development of new functional properties to filtration materials. The surface of polymeric membranes was altered by the deposition by magnetron sputtering method of thin layers of metal oxides such as copper oxide, silver oxide or titanium oxide.

Background & aims: Only limited therapeutic options are currently available for hepatocellular carcinoma (HCC), making the development of effective alternatives essential. Based on the recent finding that systemic or local hypothyroidism is associated with HCC development in humans and rodents, we investigated whether the thyroid hormone triiodothyronine (T3) could inhibit the progression of HCCs. Methods: Different rat and mouse models of hepatocarcinogenesis were investigated. The effect of T3 on tumorigenesis and metabolism/differentiation was evaluated by transcriptomic analysis, quantitative reverse transcription PCR, immunohistochemistry, and enzymatic assay. Results: A short treatment with T3 caused a shift in the global expression profile of the most aggressive preneoplastic nodules towards that of normal liver. This genomic reprogramming preceded the disappearance of nodules and involved reprogramming of metabolic genes, as well as pro-differentiating transcription factors, including Kruppel-like factor 9, a target of the thyroid hormone receptor β (TRβ). Treatment of HCC-bearing rats with T3 strongly reduced the number and burden of HCCs. Reactivation of a local T3/TRβ axis, a switch from Warburg to oxidative metabolism and loss of markers of poorly differentiated hepatocytes accompanied the reduced burden of HCC. This effect persisted 1 month after T3 withdrawal, suggesting a long-lasting effect of the hormone. The antitumorigenic effect of T3 was further supported by its inhibitory activity on cell growth and the tumorigenic ability of human HCC cell lines. Conclusions: Collectively, these findings suggest that reactivation of the T3/TRβ axis induces differentiation of neoplastic cells towards a more benign phenotype and that T3 or its analogs, particularly agonists of TRβ, could be useful tools in HCC therapy. Lay summary: Hepatocellular carcinoma (HCC) represents an important challenge for global health. Recent findings showed that systemic or local hypothyroidism is associated with HCC development. In rat models, we showed that administration of the thyroid hormone T3 impaired HCC progression, even when given at late stages. This is relevant from a translational point of view as HCC is often diagnosed at an advanced stage when it is no longer amenable to curative treatments. Thyroid hormones and/or thyromimetics could be useful for the treatment of patients with HCC.

The aim of this study was experimental and modelling analysis of the nanofiltration process used for the removal of chromium(III) ions from salt solution characterized by low pH. The experimental results were interpreted with Donnan and Steric Partitioning Pore (DSP) model based on the extended Nernst-Planck equation. In this model, one of the main parameters, describing retention of ions by the membrane, is pore dielectric constant. In this work, it was identified for various process pressures and feed compositions. The obtained results showed the satisfactory agreement between the experimental and modelling data. It means that the DSP model may be helpful for the monitoring of nanofiltration process applied for treatment of chromium tannery wastewater.

WstępW pracy przedstawiono wyniki badań florystycznych, przeprowadzonych w Dolinie Potoku Psarskiego w Będzinie. Opisano florę naczyniową badanego terenu, ze szczególnym uwzględnieniem gatunków roślin leczniczych, a także rzadkich, zagrożonych i objętych ochroną prawną.Materiał i metodyBadania terenowe prowadzono według standardowej metody florystycznej w sezonach wegetacyjnych 2011–2012. Sporządzono listę florystyczną, w której podano informacje o zagrożeniu gatunków oraz o ich ochronie prawnej. Wskazano również możliwości wykorzystania występujących na badanym obszarze roślin.WynikiW latach 2011–2012 w Dolinie Potoku Psarskiego odnotowano 295 gatunków roślin naczyniowych, a wyniki badań terenowych zestawiono z danymi literaturowymi. Na badanym obszarze nie udało się potwierdzić występowania 14 gatunków, podawanych w dwóch starszych publikacjach. 106 gatunków badanej flory uznawanych jest za rośliny lecznicze (36%), a spośród nich Farmakopea Polska X opisuje surowce pozyskiwane z 37 gatunków. Ochronie prawnej podlega 8 gatunków, a 20 uznano za rzadkie i zagrożone w województwie śląskim. Najbardziej interesującym nowym znaleziskiem florystycznym jest Iris sibirica L., uznawany dotąd za wymarły na badanym obszarze.WnioskiBadana dolina cechuje się dużą bioróżnorodnością i jest ostoją dla wielu gatunków chronionych oraz roślin regionalnie rzadkich i zagrożonych. Licznie występujące w Dolinie Potoku Psarskiego gatunki lecznicze mogą posłużyć jako naturalny rezerwuar nasion, zwłaszcza roślin rzadkich i chronionych, a także do celów naukowych.