Morphological parameters of hepatocytes in the European mole (Talpa europaea) and herb field mouse (Sylvaemus uralensis) under industrial pollution: Qualitative and quantitative assessment.

Morphological alterations of cells and tissues usually occur in biological organisms exposed to environmental contaminants, there by acting as a biomarker of environmental pollution, thus, making this study highly pertinent. The effect of industrial pollution on the qualitative and quantitative morphological parameters of hepatocytes (through histological analysis and cytomorphometry) was studied in two contrasting species of small mammals (Talpa europaea and Sylvaemus uralensis), taking into account the animal age (young and adult groups) and liver concentrations of heavy metals (Cu, Zn, Cd, Pb). Studies were performed in the regions exposed to emissions from two currently operating copper smelters: Middle Ural Copper Smelter (Middle Urals, T. europaea catching area) and Karabash Copper Smelter (Southern Urals, S. uralensis catching area). Seven morphometric parameters of hepatocytes were measured, of which two key parameters were selected by the method of principal components-the cell packing density and nuclear-cytoplasmic ratio (N/C). It was found that cell packing density in T. europaea from the impact zone decreased relative to the background area in young animals. At the same time, the differences in this parameter between the age groups from the background zone were leveled in the impact area of catching. The N/C ratio in T. europaea hepatocytes showed no correlation with either animal age or site of capture (background or impact area). In S. uralensis, both parameters, even taking into account the age, were found to be insensitive to indicate an effect of industrial pollution. Dystrophic changes (tested through histological analysis) in the liver tissue were revealed in all animal groups, but their frequency did not depend on any of the factors (age, zone) as well as the level of accumulation of toxic heavy metals (Cd, Pb). Morphometric parameters of hepatocytes have proved to be more reliable indicators of pollution, compared to the frequency of liver histopathology, due to lower subjectivity in their evaluation.

Assessment of Astaxanthin Accumulation in Hepatocytes of Atlantic Salmon Fed Different Diets Using NMR Spectroscopy.

This study aimed to assess the astaxanthin (Ax) accumulation in hepatocytes isolated from farmed Atlantic salmon fed different diets (rich marine, poor, poor with marine phospholipids (MPL) and poor with docosahexaenoic acid (DHA)). Nuclear magnetic resonance (NMR) spectroscopy was used for the Ax detection and quantification. The use of the 13C-enriched Ax allowed the assessment of short-time Ax metabolism. The substitution of fish oil and meal in fish feed on plant analogs and the addition of MPL caused further catabolism and decrease of Ax accumulation in hepatocytes from 17 to about 6 mg/kg or to almost zero in the case of DHA addition. Signals assignment of the native and 13C-enriched astaxanthin in acetone were performed using 1D and 2D NMR spectra.

Quantitative Assessment of Liver Function by Using Gadoxetic Acid-enhanced MRI: Hepatocyte Uptake Ratio.

Purpose To determine whether hepatocyte uptake ratios derived at gadoxetic acid-enhanced MRI correlate with quantitative measures of liver function and can help to identify contraindication to major hepatectomy. Materials and Methods Between August 2016 and October 2016, 50 study participants with chronic liver disease or cirrhosis underwent liver MRI at 3.0 T including T1 mapping and elastography. Liver function was quantitatively assessed by using the indocyanine green retention test (ICG R15). T1 maps were obtained by using the Look-Locker sequence before and 10 minutes after gadoxetic acid administration (0.025 mmol/kg). The relationship between ICG R15 and the following MRI parameters was evaluated: pre- and postcontrast T1 values of the liver, hepatocyte uptake ratio representing the amount of contrast media solely taken into hepatocytes, liver volume, and degree of enhancement at the common bile duct. Diagnostic performance of the hepatocyte uptake ratio to identify patients with ICG R15 greater than 20% (ie, contraindication to hepatectomy) was compared with other parameters by using areas under the receiver operating characteristic curve. Results Hepatocyte uptake ratio showed a negative correlation with ICG R15 r of -0.78 (P < .001). In participants with chronic liver disease or Child-Pugh class A, those with ICG R15 of 20% or less showed higher hepatocyte uptake ratio than those with ICG R15 greater than 20% (P < .001). Hepatocyte uptake ratios demonstrated better performance for helping to detect ICG R15 greater than 20% than did liver volume (area under the curve, 0.96 vs 0.70; P = .01). Conclusion Hepatocyte uptake ratios are negatively correlated with liver function as measured by indocyanine green retention test and provide acceptable diagnostic performance for helping to identify participants who have contraindications to major hepatectomy. © RSNA, 2018 Online supplemental material is available for this article. See also the editorial by Reeder in this issue.

Extra Pulmonary Toxicity Assessment of Gold and Silver Nanorods Following Intra Tracheal Instillation in Rats.

The gold nanorods (GNRs) and silver nanorods (SNRs) are utilized in various types of industrial and commercial applications. But, there is limited availability of extra pulmonary toxicity data regarding these nanorods. The present investigation evaluated the extra pulmonary toxicity induced by 10 and 25 nm GNRs and SNRs in rats following intra tracheal instillation. The serum biochemical analysis results have shown elevated levels of serum alanine transaminase (ALT) and serum creatinine following 1 day and 1 week post instillation. GNRs have shown greatly increased serum ALT levels at 1 day, 1 week and 1 month post exposure periods compared to SNRs and quartz (QTZ) treated rats. In case of serum creatinine levels, both GNRs and SNRs have shown similar elevated levels. Histopathology studies of rat liver tissues following exposure of GNRs and SNRs displayed that congestion of central vein, shrinkage and ballooning of hepatocytes and lymphocytic infiltration leading to degeneration after 1 week and 1 month post instillation periods. The histopathology of rat kidney tissue was showed tubular dilation, degeneration and necrosis with 10 nm SNRs and 10 nm GNRs after 1 month post instillation period. The 10 nm GNRs and SNRs have shown great changes in serum biochemical analysis and histopathological studies compared to 25 nm test nanorods. These observations suggest the size and dose dependent translocation and extra pulmonary toxicity of both GNRs and SNRs.

Assessment of magnesium influence on fatty acid content in isolated rat hepatocytes subjected to incubation.

Magnesium salts are components of many dietary supplements used in treatment or prevention of magnesium deficiency. Hypomagnesemia usually results from an improper lifestyle, including unbalanced diet. Isolated hepatocytes of animals or humans are the preferred model used to study the in vitro effects of exogenous factors on cellular metabolic changes. The aim of this study was to evaluate the content of saturated, monounsaturated and polyunsaturated fatty acids and their esters in isolated rat hepatocytes influenced by different magnesium concentrations. The isolated rat hepatocytes were used as the test material. Hepatocytes were prepared in culture medium (Hepatocyte Medium) + MgCl(2) solution to concentrations of 2 mM/dm(3) MgCl(2), 4 mM/dm(3) MgCl(2). After incubation with different concentrations of magnesium ions, changes in the content of fatty acids and their esters were found for the whole hepatocytes and hepatocyte membranes. Despite changes in the fatty acid content in the whole hepatocytes and their membranes, there were no changes in the coefficient of degree of saturation of fatty acids when different concentrations of MgCl2 were used.

Liver microsomes and S9 from rainbow trout (Oncorhynchus mykiss): comparison of basal-level enzyme activities with rat and determination of xenobiotic intrinsic clearance in support of bioaccumulation assessment.

Metabolism plays an important role in bioaccumulation of xenobiotics in fish. The applicability of trout liver microsomes and S9 fraction in bioaccumulation assessment of xenobiotics in fish was investigated in the present study. Basal-level activities of 7-ethoxyresorufin-O-dealkylase, testosterone 6beta-hydroxylase, glutathione-S-transferase, and uridine 5'-diphospho-glucuronosyltransferase in trout liver microsomes and S9 were significantly lower than those in rat liver microsomes and S9. The in vitro-to- in vivo scaling factors, which are the values of liver microsomal and S9 protein contents per unit weight of trout liver, were determined to be 38.4 +/- 5.1 (mean +/- standard deviation throughout) and 95.9 +/- 11.9 mg/g, respectively. Intrinsic clearance (CL(int)) values for a number of reference compounds obtained from trout liver S9 were lower than those from trout liver microsomes. After correction with the scaling factors, trout liver microsomes and S9 provided equivalent prediction of trout hepatic clearance (CL(H)) using the well-stirred liver model, but their CL(H) values were significantly lower than those obtained from freshly isolated trout hepatocytes. Consequently, trout liver microsomes and S9 showed poorer prediction of the bioconcentration factors of the reference compounds compared with trout hepatocytes. Unit conversion revealed that CL(int) values obtained from trout liver microsomes and S9 were 6.3 to 22.4% of those from trout hepatocytes, which explained, to a large extent, the differences in their CL(H) and bioconcentration factor prediction.

Assessment of NK cells response to hepatocyte derived chemotactic agents.

This project was aimed to examine the NK92 cells response as the CXC chemokine responder cells in rat model of liver disorder and injuries. Hepatocytes were isolated from Sprague-dawly rats and cultured on collagen type 1. Migration of NK92 cells was assessed using a 48 well micro-chemotaxis technique. Transwell chambers were positioned faced up, blocked Medium supernatant (500 microL) obtained from hepatocytes cultures were placed into the lower compartment of each Transwell. The upper compartment was filled with either 500 microL of NK92 cells. After washing, Membrane-attached cells were fixed; stained and Membrane-attached cells were counted by light microscopy and/or by size gating (9-14 microm) with an automated counter system. Human NK92 cells were attracted to recombinant human IP-10 in a concentration and time-dependent manner. NK92 cells also exhibited a chemotactic response to medium harvested from primary hepatocyte cultures. Isolated and cultured hepatocytes express several different chemokines. Although we identified that medium from hepatocyte cultures contains specific chemokines by immunoblotting, there is potential that migration assays detected yet other chemokines and other factors such as complement components. In this report, we demonstrated that hepatocytes expressed factors that were chemoattractive for human NK92 cells and that the factors must interact with the repertoire of receptors responsible for recruitment of these cells.

Immunohistochemical assessment of parafibromin in mouse and human tissues.

Parafibromin is a protein encoded by the HRPT2 oncosuppressor gene, whose mutation causes the hyperparathyroidism-jaw tumour syndrome, characterized by the occurrence of parathyroid adenoma or carcinoma, fibro-osseous jaw tumours, and renal neoplastic and non-neoplastic abnormalities. Non-morphological techniques, such as Northern and Western blotting and reverse transcriptase-PCR, indicate that parafibromin is ubiquitously expressed, but extensive immunohistochemical studies have not been performed. To increase our knowledge of the distribution and patterns of expression of parafibromin, we examined its expression and location in many different mouse and human organs by immunohistochemistry. There were no substantial differences in parafibromin expression between mouse and human. We found widespread expression of parafibromin, except in connective tissue, smooth muscle, endothelium and some other types of epithelia (colonic, urinary, tubaric, uterine, thyroid). Heterogeneity of positivity intensity and subcellular location (nuclear, nucleocytoplasmic, cytoplasmic) was found between tissues and cell types, suggesting differential functional involvement of parafibromin. Moreover, higher parafibromin expression was found in cell types, such as hepatocytes, cells of the base of gastric glands, renal cortex tubules and the pars intermedia of the hypophysis, which are characterized by different proliferative capacity, thus indicating that the cellular function of parafibromin may not be reduced only to its anti-proliferative effect.