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Areeba Ahmad, Veena Maheshwari, Arif Ahmad, Rashid Saleem and Riaz Ahmad

Observation of Esterase-Like-Albumin Activity during N'-Nitrosodimethyl amine Induced Hepatic Fibrosis in a Mammalian Model

Aim: Hepatic fibrosis (HF) is characterized by irregular growth and amassing of fibrous scar tissues in the liver causing weakened hepatocytes metabolism and protein level alterations, including albumin. Albumin with Mr~68-70 kDa is unglycosylated soluble plasma protein with various biological roles. In this study, we demonstrate ‘esterase-like activity’ of albumin during NDMA-induced HF in rats.

Material and Methods: In rats, HF was induced by weekly i.p. injections of NDMA in doses of 10 mg/kg b.wt. Sera of controls (untreated) and treated rats were processed for biochemical tests, electrophoretic profiling and in-gel esterase activity localization using α, β-naphthyl acetates. H&E staining of liver sections (~ 5 μm) was done to confirm induction of HF.

Results: NDMA satisfactorily induces hepatic fibrosis within 21 days which is also evident by significant increase in SALP, SGOT, SGPT and bilirubin levels in rats. ‘Esterase-like activity’ of albumin detected in animal sera remains stable throughout the course of treatment irrespective of other biochemical changes.

Conclusion: During pathogenesis of HF, formation of stable esterase-albumin complex may have some important role and hence, prior recommending the use of albumin as diagnostic marker we propose further investigations to elucidate the mechanism of its formation.

Open access

Bartosz Czuba, Magdalena Fituch, Slawomir Mandziuk, Barbara Jodlowska-Jedrych, Wlodzimierz Matysiak, Justyna Halasa, Franciszek Burdan, Agnieszka Korga, Magdalena Iwan, Iwona Luszczewska-Sierakowska and Jarosław Dudka

Abstract

The main side effects of the administration of doxorubicin, a widely used anticancer drug, is the generation of a reactive oxygen species (ROS) in normal cells. As a result, redox disorders and secondary oxidative stress are developed. Doxorubicin ROS generation is attributed to enzymes that are produced abundantly in hepatocytes. Oxidative stress has been a well-known risk factor of doxorubicin-related toxicity. However, in addition, according to the data collected in the last decade, changes in thyroxin status can propagate ROS generation, and, thus, initiate the doxorubicin hepatic effect. Moreover, both compounds have an impact on the cell metabolism. The aim of the study was to verify the thesis that thyroxin can modulate the effect of doxorubicin with regard to redox status and lipid metabolism disorders. In our work, we determined the ratio of NADP+/ NADPH and NAD+/NADH in liver homogenates, blood ketone bodies and triglycerides in the liver and blood in rats treated with doxorubicin and thyroxin. Our results indicate that thyroxin has an insignificant effect on NAD+/NADH, NADP+/NADPH ratios and on hepatic and blood triglycerides. Moreover, thyroxin administration normalized the level of blood ketone bodies that was disturbed by doxorubicin.

Open access

Pınar Peker Akalın, Veysel Soydal Ataseven, Doğan Fırat, Yaşar Ergün, Nuri Başpınar and Oğuzhan Özcan

Abstract

The aim of this study was to determine the ceruloplasmin (Cp) and vitamin C concentrations, the total antioxidant status (TAS), and selected biochemical parameters in dairy cows spontaneously infected with bovine leukaemia virus (BLV). Of the 27 cows included in the study, 18 animals were seropositive for enzootic bovine leukosis (EBL), whereas nine cows were seronegative and were used as controls. The serum aspartate aminotransferase (AST) (P = 0.003) and Cp concentrations (P = 0.03) decreased (65.17 ± 5.03 and 7.70 ± 0.72 respectively) in BLV-infected cows, as compared to healthy animals (100.67 ± 11.50 and 10.40 ± 0.70 respectively). A slight insignificant increase in alkaline phosphatase activity and unchanged levels of alanine aminotransferase, lactate dehydrogenase, calcium, magnesium, and TAS were demonstrated in EBL cows. As the TAS and vitamin C levels remained unchanged in EBL cows, it may be suggested that ruminants may compensate for the impaired oxidative/antioxidative balance. The results obtained also indicate that BLV may suppress AST and Cp synthesis or secretion in the liver through an unknown mechanism. The mechanism of action of BLV in hepatocytes, especially on AST and Cp, requires further investigation to elucidate the immune suppression caused by oncogenic retroviruses.

Open access

Zdzisław Zakęś, Krystyna Demska-Zakęś, Mirosław Szczepkowski, Maciej Rożyński and Elżbieta Ziomek

Abstract

The aim of the study was to determine the impact of diet and sex on the hematological and blood plasma biochemical profiles and the liver histology of pikeperch, Sander lucioperca (L.) reared in recirculating aquaculture systems (RAS) (initial mean body weight (BW) 1.35 kg). The proximate composition of the two commercial feeds used were (protein/lipid/nitrogen-free extracts) (P/L/NFE)) P505/L118/NFE294 g kg−1 (group I) and P471/L141/NFE290 g kg−1 (group II). Neither diet nor sex had a significant impact on final fish body weight (≈ 2.0 kg). Sex was noted to significantly impact glucose content (Glu – higher in males) and cholesterol (Chol – higher in females) in the blood plasma. Diet was confirmed to have a significant impact on levels of hematocrit (Ht), hemoglobin (Hb), and mean corpuscular hemoglobin (MCH), and the values of these indicators were higher in group I. Sex had a significant impact on Ht, Hb, MCH, and mean corpuscular hemoglobin concentration (MCHC), with higher values in male pikeperch. Diet and sex had significant impacts on the values of the pikeperch hepatosomatic index (HSI), hepatocyte size and that of their nuclei, and the values of the nucleocytoplasmic index (NCI).

Open access

P. A. Ahammed Shareef and Syed M. A. Abidi

Abstract

A survey on the occurrence and epidemiology of the encysted progenetic metacercariae of Euclinostomum heterostomum infection in Channa punctata in the Aligarh region of North India revealed a mean prevalence, intensity, and abundance of 18.61, 1.52, and 0.38%, respectively, during the period from April 2011 to March 2012. Liver, kidney, peritoneum, muscle, and ovary were found to be infected with this parasite, and the later three are reported for the first time in this fish species. The histopathology of the infected tissues indicated the following at the host-parasite interface: tissue damage, infiltration of immune cells into the cyst wall, chronic inflammatory responses, and granulomatous lesions. The infected liver showed degeneration of hepatocytes, cytoplasmic vacuolation, nuclear alterations, mallory body formation, fibrosis, and necrosis. The pathology of the infected kidney included distortion and dilation of renal tubules, vacuolar degeneration, hypertrophy and hyperplasia of tubular epithelial cells, occlusion of tubules, fibrosis, hemorrhage, and congestion of glomeruli. The infected muscle demonstrated comparatively fewer pathological changes confined only to the circumference of the cyst wall. The ovary displayed the least changes. The conclusions drawn from the study are that the large metacercarial cysts formed by E. heterostomum in the vital organs of the economically important fish C. punctata could result in the impairment of fish physiology and health, thereby affecting their productivity and quality for human consumption.

Open access

Shahbazi S, Mahdian R, Karimi K and Mashayekhi A

Abstract

Coagulation factor VII (FVII) is a key enzyme of the extrinsic coagulation cascade that is predominantly produced by hepatocytes. The F7 gene mutations cause FVII deficiency with considerable molecular and phenotypic heterogeneity. We characterized the molecular alterations of the F7 gene and their corresponding mRNA transcripts in Iranian patients from eight unrelated families. The mutations were detected by polymerase chain reaction (PCR)-sequencing of all F7 gene exons, their flanking intronic sequences, as well as their corresponding cDNA fragments. Homozygous P303T, C91S and R304Q mutations were detected in patient 2, patient 5, and patient 6, respectively. Patient 7 was a compound heterozygote for S282R and H348R and patient 8 was a compound heterozygote for R304Q and IVS7+7A>G mutations. Furthermore, our investigation revealed three heterozygous individuals, patient 1 and patient 3 with the A244V mutation who were symptomatic and patient 4 with V(–39)I mutation who was also asymptomatic. The F7 mRNA expression analysis revealed that, except the transcript of V(–39)I, other mutation-harboring transcripts were expressed at detectable levels. In conclusion, this report reinforces the genetic and phenotypic heterogeneity of FVII deficiency. The findings of the mRNA study implied that decreased FVII protein activity subsequent to missense mutations does not completely reflect the degradation of mutation-harboring mRNA.

Open access

Hang Zhang

Kohara K, Katsume A, et al . Pathogenesis of hepatitis C virus infection in Tupaia belangeri. J Virol, 2010, 84(1): 303-311. 5 Su J, Yan R, Gan Y, et al . Study on hepatitis B virus infection in adult tupaia. Chinese Journal of Pathology, 1987, 16 (2): 103-105. 6 Walter E, Keist R, Niederost B, et al . Hepatitis B virus infection of tupaia hepatocytes in vitro and in vivo. Hepatology, 1996, 24(1): 1-5. 7 Yan RQ, Su J J, Huang DR, et al. Human hepatitis B virus and hepatocellular carcinoma. I. Experimental infection of tree shrews with hepatitis

Open access

Sendi Montanic, Michela Terdoslavich, Uros Rajcevic, Luigina De Leo, Serena Department of Medical Sciences, University of Trieste, Cattinara Hospital, Trieste, Italy, Vladka Curin Serbec and Sabina Passamonti

Background. Bilitranslocase (TC 2.A.65.1.1) is a bilirubin-specific membrane transporter, found on absorptive (stomach and intestine) and excretory (kidney and liver) epithelia and in vascular endothelium. Polyclonal antibodies have been raised in rabbits in the past, using a synthetic peptide corresponding to AA65-77 of rat liver bilitranslocase, as an antigen. Affinity-purified antibodies from immune sera have been found to inhibit various membrane transport functions, including the bilirubin uptake into human hepatocytes and the uptake of some flavonoids into human vascular endothelial cells. It was described by means of immunohistochemistry using polyclonal antibodies that bilitranslocase expression is severely down-regulated in clear cell renal carcinoma. The aim of our work was development and characterization of high-affinity, specific mAbs against bilitranslocase, which can be used as a potential diagnostic tool in renal cell carcinoma as well as in a wide variety of biological assays on different human tissues.

Materials and methods. Mice were immunized with a multi-antigen peptide corresponding to segment 65-75 of predicted primary structure of the bilitranslocase protein. By a sequence of cloning, immune- and functional tests, we aimed at obtaining a specific monoclonal antibody which recognizes a 37 kDa membrane protein, and influences the transport activity of bilitranslocase.

Results. On the basis of previous results, specific IgM monoclonal antibodies were produced in BALB/c mice, in order to further improve and extend the immunological approach to the study of bilitranslocase in renal cancer cells as well as to develop its potential diagnostics use.

Conclusions. In this article we show an immunological approach, based on newly developed monoclonal antibodies, to a detailed biochemical and functional characterization of a protein whose gene and protein structure is still unknown. We were able to demonstrate our novel mAb as a tumor marker candidate of renal cell carcinoma, which may prove useful in the diagnostic procedures.

Open access

Isaac J. Asiedu-Gyekye, Abdulai Seidu Mahmood, Charles Awortwe and Alexander K. Nyarko

Abstract

Polyhexamethylene biguanide (PHMB) is an antiseptic with antiviral and antibacterial properties used in a variety of products including wound care dressings, contact lens cleaning solutions, perioperative cleansing products, and swimming pool cleaners. There are regulatory concerns with regard to its safety in humans for water treatment. We decided to assess the safety of this chemical in Sprague-Dawley rats. PHMB was administered in a single dose by gavage via a stomach tube as per the manufacturer’s instruction within a dose range of 2 mg/kg to 40 mg/kg. Subchronic toxicity studies were also conducted at doses of 2 mg/kg, 8 mg/kg and 32 mg/kg body weight and hematological, biochemical and histopathological findings of the major organs were assessed. Administration of a dose of 25.6 mg/kg, i.e. 1.6 mL of 0.4% PHMB solution (equivalent to 6.4×103 mg/L of 0.1% solution) resulted in 50% mortality. Histopathological analysis in the acute toxicity studies showed that no histopathological lesions were observed in the heart and kidney samples but 30% of the animals had mild hydropic changes in zone 1 of their liver samples, while at a dosage of 32 mg/kg in the subchronic toxicity studies, 50% of the animals showed either mild hepatocyte cytolysis with or without lymphocyte infiltration and feathery degeneration. Lymphocyte infiltration was, for the first time, observed in one heart sample, whereas one kidney sample showed mild tubular damage. The acute studies showed that the median lethal dose (LD50) is 25.6 mg/kg (LC50 of 1.6 mL of 0.4% PHMB. Subchronic toxicological studies also revealed few deleterious effects on the internal organs examined, as seen from the results of the biochemical parameters evaluated. These results have implications for the use of PHMB to make water potable.

Open access

Mirdza Apsīte, Nadežda Bērziņa and Natālija Basova

Abstract

Lohman Brown chickens with age from the 1 st to 35 th day received the food with high doses of selenium (Se1 mg/kg), copper (Cu100 mg/kg), or both elements (Se1 + Cu100). Live weight increase of all three experimental chicken groups was by 9.3, 12.9 and 8.1%, respectively, in comparison with the control. The concentration of selenium in the blood of the Se1 group chickens was by 45.5, in liver by 63.4 and in kidney by 19.7% higher that in organs of control group chickens. Selenium accumulation in organs of Se1 group chickens was highly correlated with increase of glutathionperoxidase activity in blood (r = 0.90) and in liver (r = 0.85) and with decrease of glutathione concentration in liver. In Cu100 group chickens, copper concentration increased by 11.7 in blood, in liver by 23.7, and in kidney by 19.9%. Together with more intensive excretion of glutathione from hepatocytes, copper concentration in bile increased by 17.7% compared to that in control group chickens. Also wing feathers participated in the regulation of copper homeostatic balance, as copper concentration in feathers increased by 66.7%. The concentration of malondialdehide in liver of chickens from all groups was similar (43.5-45.2 μmol·g-1 wet wt.), indicating that overload of selenium and copper did not cause profuse production of oxyradicals in the organism. Increased accumulation of selenium and copper in chickens influenced biochemical regulation of iron, zinc and cadmium deposition in liver, kidney, tibia and feather, changing the relations between Se and Fe, Se and Cd, Cu and Fe, Cu and Zn, and Cu and Cd concentrations. The analysis indicates increased tolerance of chicken to loads of selenium (1 mg/kg) and copper (100 mg/kg) doses