Hematological, immunological, and polyamines alterations in the concomitant occurrence of Fasciola gigantica and hepatic leiomyoma in cattle.

No previous studies have investigated the polyamines alterations during fascioliasis due to F. gigantica in ruminants. This study was therefore carried out to find out the possible relationship between the extent of liver destruction and leiomyoma and some hematological and immunological parameters and polyamines alterations in F. gigantica infection. Fifty cattle with liver fascioliasis and fifteen healthy cattle were selected for the study. For the histopathological study, liver tissue samples were stained with hematoxylin and eosin (H&E) and Masson's Trichrome methods. The leiomyoma suspected specimens were immunohistochemically stained for smooth muscle actin and desmin. Different hematological parameters were investigated in infected and non-infected animals. Furthermore, levels of putrescine, spermidine, and spermine were measured in homogenized liver samples. Serum IL-4 and TNF-α levels were also evaluated. By histological examination, the lesions were noted in all the infected specimens. These lesions were varied from leiomyoma, chronic catarrhal cholangitis, arteriosclerosis, telangiectasia, and fresh migratory tunnels filled with RBC and eosinophils. Comparison of hemogram results between infected and non-infected groups revealed a significant decrease in red blood cell counts (RBC), mean corpuscular hemoglobin concentration (MCHC), and platelet count (PLT) in infected animals. Also, a significant elevation in mean corpuscular volume (MCV) concentration was detected in infected animals. The putrescine and spermine levels of the infected animals were significantly higher than the non-infected animals. Although spermidine was increased in infected livers, its elevation was not significant. Based on the results, the level of IL-4 and TNF-α was not significantly changed in infected animals. In conclusion, the concurrent occurrence of leiomyoma and fascioliasis due to F. gigantica and polyamines elevation (putrescine and spermine) is reported for the first time. The role of polyamines in the concurrent occurrence of leiomyoma and fascioliasis is an area for future research.

Automatic classification of schizophrenia patients using resting-state EEG signals.

Schizophrenia is one of the serious mental disorders, which can suspend the patient from all aspects of life. In this paper we introduced a new method based on the adaptive neuro fuzzy inference system (ANFIS) to classify recorded electroencephalogram (EEG) signals from 14 schizophrenia patients and 14 age-matched control participants. Sixteen EEG channels from 19 main channels that had the most discriminatory information were selected. Possible artifacts of these channels were eliminated with the second-order Butterworth filter. Four features, Shannon entropy, spectral entropy, approximate entropy, and the absolute value of the highest slope of autoregressive coefficients (AVLSAC) were extracted from each selected EEG channel in 5 frequency sub-bands, Delta, Theta, Alpha, Beta, and Gamma. Forty-six features were introduced among the 640 possible ones, and the results included accuracies of near 100%, 98.89%, and 95.59% for classifiers of ANFIS, support vector machine (SVM), and artificial neural network (ANN), respectively. Also, our results show that channels of alpha of O1, theta and delta of Fz and F8, and gamma of Fp1 have the most discriminatory information between the two groups. The performance of our proposed model was also compared with the recently published approaches. This study led to presenting a new decision support system (DSS) that can receive a person's EEG signal and separates the schizophrenia patient and healthy subjects with high accuracy.

Subcellular Organelle Toxicity Caused by Arsenic Nanoparticles in Isolated Rat Hepatocytes.

BACKGROUND: Arsenic, an environmental pollutant, is a carcinogenic metalloid and also an anticancer agent. OBJECTIVE: To evaluate the toxicity of arsenic nanoparticles in rat hepatocytes. METHODS: Freshly isolated rat hepatocytes were exposed to 0, 20, 40, and 100 µM of arsenic nanoparticles and its bulk counterpart. Their viability, reactive oxygen species level, glutathione depletion, mitochondrial and lysosomal damage, and apoptosis were evaluated. RESULTS: By all concentrations, lysosomal damage and apoptosis were clearly evident in hepatocytes exposed to arsenic nanoparticles. Evaluation of mitochondria and lysosomes revealed that lysosomes were highly damaged. CONCLUSION: Exposure to arsenic nanoparticles causes apoptosis and organelle impairment. The nanoparticles have potentially higher toxicity than the bulk arsenic. Lysosomes are highly affected. It seems that, instead of mitochondria, lysosomes are the first target organelles involved in the toxicity induced by arsenic nanoparticles.

Protective role of silymarin and D-penicillamine against lead-induced liver toxicity and oxidative stress.

This study was performed to assess hepatotoxicity and alterations in liver antioxidant defence in acute lead (Pb) exposure and the protective effects of silymarin in comparison to D-penicillamine in rats. Forty eight Albino rats were divided in eight groups and received the following treatments in a 10-day experiment - group 1: normal saline as control; group 2: 25-mg/kg Pb acetate, intraperitoneally (IP) for the last 5 days; group 3: 100-mg/kg D-penicillamine, IP for the last 5 days; group 4: 200-mg/kg silymarin, orally for 10 days; and groups 5, 6, 7 and 8: in addition to Pb, they received D-penicillamine, for the last 5 days, silymarin for 10 days, a combination of silymarin for 10 days and D-penicillamine for the last 5 days and silymarin for the last 5 days, respectively. Pb acetate exposure induced significant elevation in serum alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzyme activities in group 2 compared to control group. Significant reductions in serum total protein and albumin in all Pb-exposed groups and in serum glucose in groups 2, 6 and 8 were also observed. Liver tissue superoxide dismutase and glutathione peroxidase were significantly lower in groups 2 and 8 compared to control group. Silymarin pretreatment and D-penicillamine administration in groups 5, 7 and 8 could significantly lower ALP, ALT and AST and improve liver antioxidant enzymes. Thus, acute Pb exposure induced hepatotoxicity with suppression of liver antioxidant defence system and silymarin, as an antioxidant could alleviate this effect.

Effect of Enrofloxacin on Histochemistry, Immunohistochemistry and Molecular Changes in Lamb Articular Cartilage.

Enrofloxacin is a synthetic chemotherapeutic agent from the class of the fluoroquinolones that is widely used to treat bacterial infections. It is metabolized to ciprofloxacin in the body as active metabolite. Fluoroquinolones change in the articular cartilage, especially with high doses and more than two weeks use. So, due to relatively excessive use of enrofloxacin in mammals and similarity of lambs to human subjects with respect to skeletal activity cycles, this study was done to investigate the effects of enrofloxacin on some cellular and molecular changes in growing lamb articular cartilage to evaluate some possible mechanisms involved these changes. Twelve, 2-month-old male lambs divided into three groups: control group received only normal saline; therapeutic group received 5mg/kg enrofloxacin subcutaneously, daily, for 15 days and toxic group received 35 mg/kg enrofloxacin in the same manner as therapeutic group. Twenty four hours after the last dose, the animals were sacrificed, and their stifle joints were dissected. Sampling from distal femoral and proximal tibial extremities was done quickly for further histological and molecular studies. Collagen-п content was studied with avidin-biotin immunohistochemistry method in different groups. Expression of Sox9 and caspase-3 was evaluated by Real-time PCR. Immunohistochemical changes were included decreases of matrix proteoglycans, carbohydrates, and Collagen-п in the toxic group. Some of these changes were observed in the therapeutic group with less intensity in comparison to the toxic group. Enrofloxacin were significantly decreased (P≤0.05). Sox9 expression in therapeutic and toxic groups compared to control group. But caspase -3 expressions in the toxic group significantly increased (P≤0.0001) with a comparison to other groups, while, between control and therapeutic groups, there were no significant differences. So, it can be concluded that enrofloxacin increases apoptosis in chondrocytes and decreases their numbers. Enrofloxacin use in growing lambs even at recommended therapeutic dose is not completely safe on articular cartilage. Moreover, higher doses of enrofloxacin induce severe changes in lamb articular cartilage.

Verapamil and rifampin effect on p-glycoprotein expression in hepatocellular carcinoma.

BACKGROUND: High expression of p-glycoprotein (P-gp) has been associated with a poor prognosis in patients with hepatocellular carcinoma (HCC). It is likely that P-gp overexpression is responsible for multidrug resistance in HCC. OBJECTIVES: The aim of this study was to elucidate the effect of potent carcinogen nitrosamine with and without verapamil and rifampin drugs on P-gp expression at the mRNA level in HCC. MATERIALS AND METHODS: Four groups of rats (n = 5) were selected with different treatments and one group as control. mRNA concentration changes were monitored using quantitative PCR (QPCR). RESULTS: A significant difference was found between verapamil treated group and the control regarding the mRNA level. The mdr1a mRNA was significantly decreased in the verapamil group (P ≤ 0.001). Rifampin administrated group had a decreased level of the mdr1a mRNA compared to the control group (P ≤ 0.006). No significant changes were observed in HCC induced rats regarding the mdr1a mRNA level when treated with verapamil and rifampin. An enhanced expression of the mdr1a gene was found In the HCC induced animals when treated with drugs. CONCLUSIONS: Verapamil and rifampin were found specific and effective against P-gp expression in HCC. In conclusion, treatment efficacy of most anticancer drugs is increased in combination with verapamil and rifampin against most advanced HCC.

The pharmacokinetics of Hemiscorpius lepturus scorpion venom and Razi antivenom following intramuscular administration in rat.

Hemiscorpius lepturus (H. lepturus) is one of the most dangerous scorpions in Iran. Intramuscular administration (IM) of available Razi antivenom to H. lepturus venom is used by many of Iranian clinicians. The purpose of the current study was to investigate the efficiency of IM route for treatment of envenomed patients by H. lepturus. We compared the pharmacokinetics parameters of venom and antivenom via subcutaneous (SC) and IM administration, respectively. The blood samples were taken at various predetermined time intervals, i.e., 10, 40, 60, 180, 210, 360 and 400min following 5µg (131)I-labeled venom and 5, 10, 40, 120 and 360min following 0.2ml of (131)I-labeled antivenom administration. The radio-iodination was carried out using the chloramin-T method. The results showed that pharmacokinetic parameters of the venom were T(elimination half-life) = 103.25min; Vd/F (apparent volume of distribution) = 14.9ml/kg; Cl/F (total blood clearance) = 0.04ml/kg/min; mean resident residual time (MRT) = 244.3min, and for the antivenom T(1/2) = 628.59min, Vd = 666.66ml/kg, Cl = 0.13ml/kg/min and MRT = 1292min. A comparison of pharmacokinetic profiles indicated that the intramuscular administration was helpful in the referral less than 2hr to clinical centers but not those exceeding 3hr. Overall, the data showed that immunotherapy against H. lepturus stings was likely to be more effective through intravenous administration.

Evaluation of an oral insulin formulation in normal and diabetic rats.

AIM: As injection is not an ideal means for insulin delivery, various attempts have been made to administer insulin orally until now. The development of an oral dosage form of insulin would help diabetic patients and make the treatment more convenient. The aim of the present study is to evaluate the effectiveness of an oral insulin formulation containing polar and non-polar ingredients. MATERIALS AND METHODS: New excipient for oral insulin administration in normal and diabetic rats was evaluated by measuring blood glucose concentrations in two groups (10 rats each) of normal and streptozotocin-induced diabetic rats. Oral insulin was administrated and blood glucose was measured by glucometer at 0, 1, 2, 3 and 4 h post-feeding. The data was compared by Student's t test. RESULTS: Oral insulin formulation significantly (P<0.05) reduced blood glucose from 100 mg/dl to 33.73 mg/dl and 451.66 mg/dl to 200.83 mg/dl at 4 h in normal and diabetic rats, respectively. CONCLUSION: The novel excipient used could protect insulin from gastric and pancreatic enzymes and reduce blood glucose concentration in both healthy and diabetic rats suggesting that oral delivery of insulin is feasible in a near future.

The effect of aspartate-lysine-isoleucine and aspartate-arginine-tyrosine mutations on the expression and activity of vasopressin V2 receptor gene.

BACKGROUND: Vasopressin type 2 receptor (V2R) plays an important role in the water reabsorption in the kidney collecting ducts. V2R is a G protein coupled receptor (GPCR) and the triplet of amino acids aspartate-arginine-histidine (DRH) in this receptor might significantly influence its activity similar to other GPCR. However, the role of this motif has not been fully confirmed. Therefore, the present study attempted to shed some more light on the role of DRH motif in G protein coupling and V2R function with the use of site-directed mutagenesis. METHODS: Nested PCR using specific primers was used to produce DNA fragments containing aspartate-lysine-isoleucine and aspartate-arginine-tyrosine mutations with replacements of the arginine to lysine and histidine to tyrosine, respectively. After digestion, these inserts were ligated into the pcDNA3 vector and transformation into E. coli HB101 was performed using heat shock method. The obtained colonies were analyzed for the presence and orientation of the inserts using proper restriction enzymes. After transient transfection of COS-7 cells using diethylaminoethyl-dextran method, the adenylyl cyclase activity assay was performed for functional study. The cell surface expression was analyzed by indirect ELISA method. RESULTS: The functional assay indicated that none of these mutations significantly altered cAMP production and cell surface expression of V2R in these cells. CONCLUSION: Since some substitutions in arginine residue have shown to lead to the inactive V2 receptor, further studies are required to define the role of this residue more precisely. However, it seems that the role of the histidine residue is not critical in the V2 receptor function.

Metallothionein biosynthesis as a detoxification mechanism in mercury exposure in fish, spotted scat (Scatophagus argus).

It is of crucial importance to study on the biomarkers types to assess the specification of the pollutants and health status of marine ecosystems in environmental evaluation projects. In this respect, total metallothionein biosynthesis and mercury bioaccumulation in the liver and gills under acute mercury exposure were investigated in fish, Scat (Scatophagus argus). Spotted scat was exposed to different mercury concentrations (0, 10, 20, 30) for 24, 48, 72 h. Total MT levels were determined by enzyme-linked immunosorbent assay (ELISA) method. Mercury contents were determined through cold vapor atomic absorption spectrometry (CVAAS). Induction of MT during exposure was tissue specific, displaying different response pattern in gills and liver. Mercury accumulated in liver much higher than in gills and the latter also showed lower MT level (P<0.05). MT biosynthesis in liver showed a significant (P<0.05) increase after exposure to different mercury concentration with increase in exposure time, whereas total MT content did not significantly (P>0.05) change in gills except for 72 h exposure at 30 µg l(-1). Nonetheless, the relationship between MT biosynthesis and Mercury bioaccumulation in both tissues was significant (P<0.05). The results suggest that this form of MT in S. argus was Hg inducible and could be extended as a biomarker of mercury pollution in marine ecosystems.

Comparison of the prophylactic effect of silymarin and deferoxamine on iron overload-induced hepatotoxicity in rat.

In pathologic conditions or poisoning states, iron overload can affect different tissues including liver. In this study, the prophylactic effect of deferoxamine and silymarin was compared in decreasing experimental iron-overload-induced hepatotoxicity in rats. The study was done in six groups of rats, which received drugs q2 days for 2 weeks. The rats in groups 1 to 6 received drugs, respectively: normal saline, iron dextran, iron dextran + deferoxamine (intraperitoneally), iron dextran + silymarin (orally), iron dextran + silymarin (intraperitoneally), and iron dextran + deferoxamine (intraperitoneally) + silymarin (intraperitoneally). At the end of the study, blood was collected, and serum was separated for laboratory tests. The liver of rats was separated for iron measuring and tissue processing. The serum iron concentration and the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity were determined. The numbers of necrotic hepatocytes were counted as quantity index tissue injury in light microscopic examination. The mean of serum and liver iron in group 2 was significantly greater than group 1. Liver iron was significantly decreased in other groups except group 4. Also serum iron was decreased in groups 3 to 6 compared to group 2 (nearly 400%). ALT activity in group 3 and AST activity in group 5 were significantly lesser than in other groups. The mean of necrotic hepatocytes in group 2 was significantly increased in comparison to group 1. This elevation was significantly prevented by deferoxamine and silymarin. The result of the present study shows that silymarin has a protective effect similar to deferoxamine on iron overload-induced hepatotoxicity.