Farzad Asadi

The antioxidative effect of Iranian Mentha pulegium extracts and essential oil in sunflower oil

The aim of the present study was to evaluate antioxidative activities of the essential oil, methanol and water extracts of Iranian pennyroyal in vegetable oil during storage. Different concentrations (0, 200, 400, 600, 800 and 1000 ppm) of essential oil, water and methanol extracts and beta-hydroxy toluene (BHT; 200 ppm) were added to sunflower oil emulsion in the presence of cupric ions and incubated for 7 days at 60 degrees C. Peroxide values (PVs) and thiobarbituric acid reacting substances (TBARS) levels were measured in each day up to day of seven. Furthermore, antioxidant capacity of the essential oil and extracts were determined using DPPH and beta-carotene-linoleic acid methods. Values were compared among groups in each incubation time points using ANOVA. Results showed that DPPH and beta-carotene-linoleic acid assay findings on the Mentha pulegium extracts were comparable to those found on BHT. Furthermore, in all incubation time points, M. pulegium extracts lowered PVs and TBARS levels when compared to the control (p<0.001). In this respect, water extract was more potent than the methanol extract. Essential oil did not show considerable antioxidative effect. It seems that water extract of M. pulegium is a potent antioxidant which makes it as a potential antioxidant for oil and oily products during storage.

Palmitate-Activated Macrophages Confer Insulin Resistance to Muscle Cells by a Mechanism Involving Protein Kinase C θ and ε

Background Macrophage-derived factors contribute to whole-body insulin resistance, partly by impinging on metabolically active tissues. As proof of principle for this interaction, conditioned medium from macrophages treated with palmitate (CM-PA) reduces insulin action and glucose uptake in muscle cells. However, the mechanism whereby CM-PA confers this negative response onto muscle cells remains unknown. Methodology/Principal Findings L6-GLUT4myc myoblasts were exposed for 24 h to palmitate-free conditioned medium from RAW 264.7 macrophages pre-treated with 0.5 mM palmitate for 6 h. This palmitate-free CM-PA, containing selective cytokines and chemokines, inhibited myoblast insulin-stimulated insulin receptor substrate 1 (IRS1) tyrosine phosphorylation, AS160 phosphorylation, GLUT4 translocation and glucose uptake. These effects were accompanied by a rise in c-Jun N-terminal kinase (JNK) activation, degradation of Inhibitor of κBα (IκBα), and elevated expression of proinflammatory cytokines in myoblasts. Notably, CM-PA caused IRS1 phosphorylation on Ser1101, and phosphorylation of novel PKCθ and ε. Co-incubation of myoblasts with CM-PA and the novel and conventional PKC inhibitor Gö6983 (but not with the conventional PKC inhibitor Gö6976) prevented PKCθ and ε activation, JNK phosphorylation, restored IκBα mass and reduced proinflammatory cytokine production. Gö6983 also restored insulin signalling and glucose uptake in myoblasts. Moreover, co-silencing both novel PKC θ and ε isoforms in myoblasts by RNA interference, but not their individual silencing, prevented the inflammatory response and restored insulin sensitivity to CM-PA-treated myoblasts. Conclusions/Clinical Significance The results suggest that the block in muscle insulin action caused by CM-PA is mediated by novel PKCθ and PKCε. This study re-establishes the participation of macrophages as a relay in the action of fatty acids on muscle cells, and further identifies PKCθ and PKCε as key elements in the inflammatory and insulin resistance responses of muscle cells to macrophage products. Furthermore, it portrays these PKC isoforms as potential targets for the treatment of fatty acid-induced, inflammation-linked insulin resistance.

Serum and Cerebrospinal Fluid Antioxidant Activity and Lipid Peroxidation in Guillain–Barre Syndrome and Multiple Sclerosis Patients

ABSTRACT Oxidative stress through the changes in the levels of reactive oxygen species and antioxidative parameters can cause various neurological disorders. The aim of the present study was to show antioxidant activity (AOA) and malondialdehyde (MDA) levels in affected people with Guillain–Barre syndrome (GBS) and multiple sclerosis (MS). A total of 15 GBS patients, 13 MS patients, and 15 age and sex matched controls were included in this study. MDA and AOA values were determined in both cerebrospinal fluid (CSF) and serum, spectrophotometrically. We have shown an increase in the values of MDA in the CSF of both GBS and MS patients (0.32 ± 0.073 and 0.22 ± 0.06 μmol/L) compared to the control (undetectable levels). Furthermore, a significant decrease in the serum MDA levels was shown in both GBS and MS patients (0.81 ± 0.18 and 0.73 ± 0.18 μmol/L) when compared to the control (1.7 ± 0.46 μmol/L). A decrease was shown for serum AOA in both GBS (1.7 ± 0.21 mmol/L) and MS patients (2.6 ± 0.62 mmol/L) when compared to the control (3.2 ± 0.17 mmol/L). However, a significant increase in the values of CSF AOA was shown in both MS and GBS patients (1.47 ± 0.19 and 1.42 ± 0.26 mmol/L) compared to the control (0.71 ± 0.19 mmol/L). An imbalance between the levels of AOA and MDA in both CSF and serum can be followed in both MS and GBS patients.

Molecular considerations for development of phage antibody libraries

Nowadays, phage display libraries are used as robust tools for discovery and evolution of peptide/protein based drugs as well as targeting molecules, in particular monoclonal antibodies (mAbs) and its fragments (i.e., scFvs, Fabs, or bivalent F(ab′)2). Phage display technology, as a molecular diversity approach, enables selection of antibody fragments (e.g., scFv/Fab) with high affinity, specificity and effector functions against various targets. However, such selection process itself is largely dependent upon various molecular factors such as methods for construction of phage library, phage/phagemid vectors, helper phage, host cells and biopanning processes. The current review article provides important molecular considerations for successful development of phage antibody libraries that may be used as a platform for translation of antibody fragments into viable diagnostic/therapeutic reagents.

Effect of long-term optional ingestion of canola oil, grape seed oil, corn oil and yogurt butter on serum, muscle and liver cholesterol status in rats.

The aim of the present study was to determine the effect of long-term optional intake of vegetable oils (canola, grape seed, corn) and yogurt butter on the serum, liver and muscle cholesterol status. Twenty-five male Wistar rats were randomly categorized into five groups (n=5) as follows: control, canola oil, grape seed oil, corn oil and manually prepared yogurt butter. In each group, 24h two bottle choice (oil and water) tests were performed for 10 weeks. Serum cholesterol values showed a trend to decrease in grape seed oil, corn oil and yogurt butter groups compared to the control. Optional intake of yogurt butter made a significant increase in HDL-C values (42.34+/-9.98 mg/dL) yet decrease in LDL-C values (11.68+/-2.06 mg/dL) compared to the corresponding control (19.07+/-3.51; 30.96+/-6.38 mg/dL, respectively). Furthermore, such findings were concomitant with a significant decrease in the liver TC levels (1.75+/-0.31 mg/g liver) and an increase in the muscle TC levels (1.85+/-0.32 mg/g liver) compared to the corresponding control (2.43+/-0.31; 0.94+/-0.14 mg/g liver, respectively). Optional intake of manually prepared yogurt butter has more beneficial effects on serum lipoprotein cholesterol values with some alterations in the liver and muscle cholesterol states than the vegetable oils.

Selection of potential therapeutic human single-chain Fv antibodies against cholecystokinin-B/gastrin receptor by phage display technology.

Background and ObjectiveGastric/gastrointestinal cancers are associated with high mortality worldwide. G-protein coupled receptor (GPCR) superfamily members such as gastrin/cholecystokinin-B receptor (CCK-BR) are involved in progression of gastric tumors, thus CCK-BR is considered as a potential target for immunotherapy. However, production of functional monoclonal antibodies (mAbs) against GPCR seems to be very challenging, in part due to its integration in cell membranes and inaccessibility for selection. To tackle this problem, we implemented phage display technology and a solution-phase biopanning (SPB) scheme for production of mAbs specific to the native conformation of CCK-BR.MethodsTo perform the SPB process, we utilized a synthetic biotinylated peptide corresponding to the second extracellular loop (ECL2) of CCK-BR and a semi-synthetic phage antibody library. After enzyme-linked immunosorbent assay (ELISA) screening, the CCK-BR specificity of the selected single-chain variable fragments (scFvs) were further examined using immunoblotting, whole-cell ELISA, and flow cytometry assays.ResultsAfter performing four rounds of selection, we identified nine antibody clones which showed positive reactivity with the CCK-BR peptide in an ELISA assay. Of these, eight clones were unique scFv antibodies and one was a VL single domain antibody. Specificity analysis of the selected scFvs revealed that five of the selected scFvs recognized a denatured form of CCK-BR, while the majority of the selected scFvs were able to recognize the native conformation of CCK-BR on the surface of human gastric adenocarcinoma cells and cervical carcinoma HeLa cells.ConclusionFor the first time, we report on the establishment of a diverse panel of scFv antibody fragments that are specific to the native conformation of CCK-BR. Based on these results, we suggest the selected scFv antibody fragments as potential agents for diagnosis, imaging, targeting, and/or immunotherapy of cancers that overexpress CCK-BR.

Isolation of β -glucan from the cell wall of Saccharomyces cerevisiae

β-Glucan, one of the major cell wall components of Saccharomyces cerevisiae (S. cerevisiae), has been found to enhance immune functions. At present study, we developed an optimal procedure to extract and purify β-glucan. At first, yeast cells were grown in sabouraud dextrose agar and then cultured in yeast extract–peptone–glucose (YPG) broth. After incubation, cells were harvested, washed and disrupted by means of sonication method. The obtained cell walls were used to prepare alkali-soluble β-glucan (glucan-S1). In this regard, 2% sodium hydroxide (NaOH) and 3% acetic acid were used in alkaline–acid extraction, respectively. This preparation contained 2.4% protein. In the next step, DEAE sephacel chromatography was used to remove remaining proteins (glucan-S2). Subsequently this preparation was applied into concanavalin-A sepharose column to remove manann. Finally, β-glucan free of mannoprotein complexes was prepared (glucan-S3).

Assessment of dermal exposure and histopathologic changes of different sized nano-silver in healthy adult rabbits

The purpose of this study is to evaluate the dermal toxicity (Irritation/Corrosion) of three sizes of nanosilver particles (10, 20 and 30 nm) during 3 min, 1 and 4 hours according to the OECD/OCDE guideline Histopathological effects in secondary organs from liver, kidney, heart, spleen and brain 14 day post dermal administration are also reported. 10 and 20 nm Ag nanoparticles treated group showed well defined dermal erythema and oedema. Histopathological findings of 10 and 20 nm (4 hours exposure) on 14-day post dermal administration showed hyperkeratosis, acanthosis, hair-filled follicles and papillomatosis in an irregular epidermis, fibrosis, hyperemia, erythema, intracellular oedema and hyalinisation of collagen in dermis of skin. Liver revealed midzonal and periacinar necrosis, portal mononuclear infiltration, liver fatty change, liver congestion and hyperemic central vein. Splenic red pulp congestion and white pulp hyperreactivity, splenic trabeculae and sinusoidal congestion and hyaline change were found in spleen. Fatty degeneration in some cardiovascular cells and subendocardial hemorrhage without inflammation was perceived. Picnotic appearance of pyramidal neurons in the brain cortex, gliosis and mild perineuronal oedema ischemic cell change and hyperemic meninges was observed in brain. Our research concluded that dermal exposure to lesser sizes of silver nanoparticles is more disastrous than greater ones.

Serum electrolyte and nonelectrolyte status in freshwater juvenile Persian sturgeon Acipenser persicus.

Status of serum electrolyte and nonelectrolyte variables can be used for managing sturgeon species cultured in freshwater or living in seawater. The aim of the present study was to evaluate serum biochemical variables in clinically healthy juvenile Persian sturgeon Acipenser persicus cultured in freshwater. Serum samples from 11 females and 10 males were analyzed, and levels (mean +/- SD) of the following variables were compared between sexes: glucose (Glc; 5.58 +/- 1.25 mmol/L for females and 8.56 +/- 1.80 mmol/L for males), total cholesterol (TC; 2.50 +/- 0.45 and 2.40 +/- 0.65 mmol/L), triglyceride (TG; 7.13 +/- 2.68 and 5.14 +/- 1.27 mmol/L), blood urea nitrogen (BUN; 1.28 +/- 0.2 and 1.01 +/- 0.2 mmol/L), total protein (TOP; 55.84 +/- 8.77 and 41.44 +/- 8.62 g/L), inorganic phosphate (P(i); 6.19 +/- 1.02 and 5.23 +/- 0.49 mmol/L), calcium (Ca; 2.80 +/- 0.43 and 2.63 +/- 0.32 mmol/L), magnesium (Mg; 0.9 +/- 0.23 and 0.99 +/- 0.22 mmol/L), sodium (Na; 152.80 +/- 13.81 and 156.38 +/- 12.67 mmol/L), potassium (K; 2.64 +/- 0.58 and 2.27 +/- 0.39 mmol/L), and chloride (Cl; 143 +/- 13.85 and 151.67 +/- 21.08 mmol/L). There were no differences in TC, Ca, Mg, Na, K, or Cl between sexes. The Glc value was lower in female Persian sturgeon than in males, whereas the values of TG, BUN, TOP, and P(i) were higher in females than in males. Freshwater adaptation may affect serum ion concentrations in juvenile Persian sturgeon.

Serum lipid and lipoprotein profile of Asian tortoise (Agrionemys horsfieldi) in prehibernation state

The Asian tortoise (Agrionemys horsfieldi), which now has a worldwide distribution, originates from Iran, Pakistan, West China, Kazakstan, Turkmenia, and Russia. Because of the increasing popularity of this animal as a domestic pet and its presence in many zoological gardens, knowledge of its physiological characteristics at different stages of life is important. In this regard, serum lipid and lipoprotein profiles of Asian tortoise were measured just before hibernation. Blood samples were collected from the jugular vein of 16 tortoises of both sexes. Total cholesterol (TC), phospholipid (PL), high-density lipoprotein cholesterol (HDL-C) were measured chemically, very-low-density lipoprotein cholesterol (VLDL-C) was calculated as an estimation of one fifth of triglyceride (TG), low-density lipoprotein cholesterol was determined on the basis of Friedewalds formula, and total lipid was calculated from the summation of TG, TC, and PL. PL and TC comprised 79 and 21% of total lipid in the male and 85 and 14% in the female, respectively. TG level was low in both sexes. Moreover, approximately 86 and 12% of TC were carried through LDL and HDL, respectively. Before hibernation, PL and LDL-C are the major lipid and cholesterol fractions of serum, respectively.