Iranian Journal of Pharmaceutical Research
Volume:10 Issue: 4, Autumn 2011

Mycotoxins are natural secondary fungal metabolites produced on agricultural commodities. Mould and mycotoxin contamination may occur before harvesting, between harvesting and drying, and during storage. FAO has estimated that approximately 25% of the world’s crops are contaminated by molds and affected by mycotoxins, and the estimated loss extends to billions of dollars. Since fungi have a widespread distribution in the environment, mycotoxins are considered to be one of the most important contaminants in foodstuffs and feedstuffs. More than 500 different mycotoxins are known, but only a few present considerable food safety hazards. Aspergillus, Fusarium, and Penicillium are the natural fungal flora associated with foods. The most important mycotoxins are aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, fumonisins, and patulin. The common occurrence of mycotoxins in foodstuffs and feedstuffs poses a threat to the humans and animals health. Mycotoxins have acute toxic, mutagenic, carcinogenic, teratogenic, estrogenic, and immunotoxic effects in humans and animals. Despite occurrence of mycotoxin contamination of agricultural products in the developed countries, mycotoxin exposure have greatly been reduced in these populations due to the presence of a legislatively regulated food processing and marketing system and the application of different strategies. However, in the developingcountries, much of the population relies on subsistence farming or on unregulated local markets. Therefore, regulatory limits for major mycotoxin classes and selected individual mycotoxins have been established in many countries to protect the consumer from the harmful effects of these toxins. The requirement to apply these regulatory limits has prompted development of analytical methods for the identification and quantification of mycotoxins in different foodstuffs and feedstuffs.

Non-steroidal anti-inflammatory drugs (NSAIDs) are the competitive inhibitors of cyclooxygenase (COX), the enzyme which mediates the bioconversion of arachidonic acid to inflammatory prostaglandins (PGs). Their use is associated with the side effects such as gastrointestinal and renal toxicity. The therapeutic anti-inflammatory action of NSAIDs is produced by the inhibition of COX-2, while the undesired side effects arise from inhibition of COX-1 activity. Thus, it was though that more selective COX-2 inhibitors would have reduced side effects. Based upon a number of selective COX-2 inhibitors (rofecoxib, celecoxib, valdecoxib etc.) were developed as safer NSAIDs with improved gastric safety profile. However, the recent market removal of some COXIBs such as rofecoxib due to its adverse cardiovascular side effects clearly encourages the researchers to explore and evaluate alternative templates with COX-2 inhibitory activity. Recognition of new avenues for selective COX-2 inhibitors in cancer chemotherapy and neurological diseases such as Parkinson and Alzheimer’s diseases still continues to attract investigations on the development of COX-2 inhibitors. This review highlights the various structural classes of selective COX-2 inhibitors with special emphasis on their structure-activity relationships.

A new sensitive, simple and rapid method for determination of methotrexate (MTX) was developed based on quenching effects of MTX on the fluorescence intensity of Tb3+-1,10-phenanthroline complex. The fluorescence intensity was measured with excitation wavelength of 300 nm and emission wavelength of 545 nm and the quenched fluorescence intensity is proportional to the concentration of MTX in Tris-HCl buffer solution with a pH of 7.0. The effects of pH, time, order of addition of the reagents, temperature and the concentrations of Tb3+, buffer and 1,10-phenanthroline were investigated and optimized. The obtained linear range for the determination of MTX was 0.02-10 μg/mL. The detection limits (signal: noise = 3) was 0.015 μg/mL and the relative standard deviation for replicated determinations of 1 μg/mL of MTX was 1.9%. The proposed method is a simple, practical and relatively free from interference effects and was successfully applied to assess MTX in urine, serum and samples of an injection solution.

In the present study solid dispersions of the antifungal drug Ketoconazole were prepared with Pluronic F-127 and PVP K-30 with an intention to improve its dissolution properties. Investigations of the properties of the dispersions were performed using release studies, Differential scanning calorimetery (DSC), X-ray powder diffraction (XRD) and Fourier transform infrared (FTIR). The results obtained showed that the rate of dissolution of Ketoconazole was considerably improved when formulated in solid dispersions with PVP K-30 and Pluronic F-127 as compared with pure drug and physical mixtures. The results from DSC and XRD studies showed the transition of crystalline nature of drug to amorphous form, while FTIR studies demonstrated the absence of drug-carriers interaction.

Vitex agnus-castus L. is a medicinal plant which is used in several dosage forms for women hormonal disorders and standardized according to the iridoids or flavonoids content. Aucubin, an iridoid glycoside, considered as a marker in some formulations. In this research, a thin layer chromatographic method with densitometric detection has been developed for quantitative determination of aucubin in chaste tree fruits. Chromatographic separation was performed using silica gel high performance thin-layer chromatography (HPTLC) plates with ethyl acetate-methanol-water 77: 15: 8 as mobile phase. Chromatograms were visualized using p-dimethylaminobenzaldehyde as reagent. Aucubin RF-value was about 0.5 and spots were scanned at 580 nm through a mercury lamp. By using this method, the amount of aucubin was found 43.5 mg/100 g of dried plant fruits. The method was validated for selectivity, linearity (r2 = 0.997, 20-100 µg/mL), precision (intra-day < 4.9, inter-day < 7.2) and accuracy measured via determination of recovery (95-98%). The limit of detection and limit of quantization were found 6.6 and 20 µg/mL, respectively. This methodology was found to be precise with respect to the validation parameters. It is simple and convenient and could be applicable to the routine determination of aucubin in different Vitex agnus-castus L. samples.

In a search for new leads towards potent antimicrobial agents, an array of novel (2E)-ethyl-2-(2-(2,4-dinitrophenyl)hydrazono)-4-(naphthalen-2-yl)-6-arylcyclohex-3-ene carboxylates 17-24 were synthesized and characterized through their melting point, elemental analysis, MS, FT-IR, one-dimensional NMR (1H, D2O exchanged 1H and 13C), two dimensional HOMOCOR and HSQC spectroscopic data. In-vitro microbiological evaluations were carried out for all the newly synthesized compounds 17-24 against clinically isolated bacterial strains namely Salmonella typhii, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, β-Hemolytic streptococcus and Micrococcus luteus and also fungal strains namely Aspergillus flavus, Aspergillus niger, Mucor, Rhizopus and Microsporum gypseum and finally, the results of their structure activity relationship were discussed. The obtained results can be used as the key step for the building of novel chemical compounds with interesting antimicrobial profiles comparable to that of the standard drugs.

A series of new 2-(phenylthio) benzoylaryl hydrazones were synthesized by acid-catalyzed condensation of hydrazide 3 with corresponding aldehydes. The chemical structures of the compounds were elucidated by FT-IR, 1H-NMR and Mass spectra. All newly synthesized compounds were evaluated for their antimycobacterial activities against Mycobacterium tuberculosis H37Rv using the microplate alamar blue assay (MABA). Compounds 4f (5-Nitro-2-furyl analogue) and 4g (5-Nitro-2-thienyl analogue) showed antimycobacterial activity with IC90, 7.57 and 2.96 µg/mL, respectively.

A series of 2-methylthio-1,4-dihydropyrimidine derivatives (II) were synthesized in good yields by alkylation of 1,2,3,4-tetrahydropyrimidines (I) with methyl iodide in the presence of pyridine. Their structures were confirmed by elemental analysis, IR and 1H NMR spectra. The compounds were tested for analgesic activity by acetic acid induced writhing method. Compounds IIh, IIe, IIk and IIl showed excellent to good analgesic activity. Other compounds showed moderate analgesic activity. The observed analgesic activity is mainly because of inhibition of the peripheral pain mechanism by the title compounds.

Sulfonamides are the first effective chemotherapeutic agents used for several years to cure or prevent systemic bacterial infections. In addition, this agents showed anti-carbonic anhydrase and cause cell cycle perturbation in the G1 phase, disruption of microtubule assembly, suppression of the transcription activator Nf-Y, angiogenesis and matrix metalloproteinase (MMP). In recent years, novel synthesized sulfonamides have been introduced as antitumor, antiviral and anti-inflammatory agents. In this paper, the cytotoxic effects of 8 synthesized sulfonamides were investigated by MTT assay on HeLa, MDA-MD-468 and MCF-7 cancer cell lines. Human cancer cells were cultured and passaged in RPMI-1640 medium. Cells incubated in 96-well plates in a concentration of 1 × 105 cells/mL for 24 h, and then logarithmic concentrations (0.1 µm, 1 µm, 10 µm, 100 µm, 1mM) of each drug were prepared, added to the plates and incubated for 72 h. Cell survival was then determined using ELISA plate reader in 540 nm applying MTT assay. All tested sulfonamides showed cytotoxic effect on HeLa and MCF-7 cells in the concentration range of 100-1000 µm. These sulfonamides were cytotoxic against MDA-MB-468 cell line at a concentration of 10-100 µm and reduced the cell survival less than 50%. According to the results calculated IC50’s were as following: MDA-MB-468 < 30 µm; MCF-7 < 128 µm and HeLa < 360 µm. In conclusion, some tested sulfonamides had good cytotoxic effects against breast cancer cells, MDA-MB-468 and further investigations are needed to confirm their effects against other cells.

The great demand of people for consumption of natural additives resulted in producing natural vanillin. There are plant sources and chemical procedures for vanillin production but microbial bioconversions are being sought as a suitable alternative. In the present work, the ability to produce vanillin from isoeugenol was screened using growing cultures of various bacteria. Among the 56 strains of bacteria isolated from the soil environments of Iran, a Gram-negative rod designated as strain ISPC2 showed the capability of promoting the formation of high amounts of vanillin when grown in the presence of isoeugenol. On the basis of morphological and physiochemical characteristics and 16S ribosomal ribonucleic acid (rRNA) gene sequence analysis, the isolate was identified as Pseudomonas aeruginosa ISPC2. Vanillin formation was analyzed by GC/FID. In the presence of isoeugenol, a growing culture of P. aeruginosa ISPC2 produced 1.62 gL-1 vanillin (molar yield of 17.3%) after a 72 h reaction at 30°C and 200 rpm. This proposed procedure is an alternative approach to obtain vanillin in an environmentally friendly way. Further studies for standardization and optimization for higher yield of vanillin production, needs to be investigated.

A soil bacterium capable of degrading chitin on chitin agar plates was isolated and identified as Bacillus pumilus isolate U5 on the basis of 16S rDNA sequence analysis. In order to optimize culture conditions for chitinase production by this bacterium, a two step approach was employed. First, the effects of several medium components were studied using the Plackett-Burman design. Among various components tested, chitin and yeast extract showed positive effect on enzyme production while MgSO4 and FeSO4 had negative effect. However, the linear model proved to be insufficient for determining the optimum levels for these components due to a highly significant curvature effect. In the second step, Box-Behnken response surface methodology was used to determine the optimum values. It was noticed that a quadratic polynomial equation fitted he experimental data appropriately. The optimum concentrations for chitin, yeast extract, MgSO4 and FeSO4 were found to be 4.76, 0.439, 0.0055 and 0.019 g/L, respectively, with a predicted value of chitinase production of 97.67 U/100 mL. Using this statistically optimized medium, the practical chitinase production reached 96.1 U/100 mL.

The essential oils, isolated by hydrodistillation from the leaves, seeds and rhizomes of Heracleum sprengelianum (Wight and Arnott), collected from the Western Ghats of Peninsula India, were analyzed by gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC–MS). The antioxidant property of these oils was tested, with and without peroxidation inducer, through the egg yolk-based Thiobarbituric Acid Reactive Substances assay (TBARS assay) and in the concentrations of 50, 100, 250 and 500 mg/L. β-Pinene, 1,8-Cineole, β-Phellandrene and ρ-Cymen-8-ol were the main components of H. sprengelianum leaves, seeds and rhizomes essential oils. The oils demonstrated the antioxidant capacity in the absence of radical inducer 2, 20-azobis-(2-amidinopropane) dihydrochloride (ABAP), mainly that of H. sprengelianum at 250 and 500 mg/L, comparable in some cases to that of α-tocopherol and butylated hydroxytoluene (BHT). The presence of ABAP diminished the antioxidant ability of all tested essential oils, leaf oils of H. sprengelianum still showing the highest antioxidant capacity at 500 mg/L. At 250 and 500 mg/L for BHA, and 500 mg/L for α-tocopherol, the antioxidant capacity significantly increased in the presence of ABAP.

Lesser Periwinkle (Vinca minor L.), a member of Apocynaceae, is not only an ornamental plant with lilac-blue flowers, but also a medical plant producing an important alkaloid, vincamine, found in the leaves which shows a pronounced cerebrovasodilatory and neuroprotective activity. This plant is native to northern Spain, western France, central and southern Europe, and Caucasus. It has been recently cultivated for pharmaceutical purposes by Zardband Botanical Garden in Iran. Since the quality of herb material and alkaloid concentration is greatly influenced by environmental conditions, in this study, we report the isolation and identification of major alkaloids along with the quantification of vincamine as the pharmacologically most important component. Alkaloids from the aerial parts of V. minor were isolated and purified using different chromatographic methods. The structures of these alkaloids were determined on the basis of their physical and spectroscopic data. The concentration of vincamine was determined by high performance liquid chromatography using Tracer Excel 120 ODS A C18 column. Five indole alkaloids including vincaminorine, vincaminoreine, minovine, minovincine, and vincamine (Figure 1) were isolated from the aerial parts of V. minor. Vincamine was found to be the dominant alkaloid in this plant with the content of 0.057% of the dried plant mass. This plant may be used as a natural source for pharmaceutical purposes in Iran, due to the presence of biologically active alkaloids especially vincamine as the major alkaloid in Lesser Periwinkle cultivated.

Antioxidant effects of ethyl acetate fraction of Mentha spicata (L.) were evaluated against 4-nitroquinoline-1-oxide injected mice. For this study, experiment setup consisted of 36 albino mice of either sex divided into 6 groups: Control (25% DMSO in water), ethyl acetate fraction (EAF) alone group (80, 160 mg/Kg body weight-bwt), 4-NQO (7.5 mg/Kg bwt-IP) alone and 4-NQO + EAF. EAF and vehicles were administered orally for five consecutive days. 4-NQO (7.5 mg/Kg bwt) was injected intraperitoneally on the 6th day. After 24 h, the animals were killed; liver sample was extracted and used for bio-assay. 4-NQO alone treated group decreased (27-60%) the antioxidant activities and promoted lipid peroxidation (LPO-60%) over their respective control values. Pretreatment with EAF, at the maximum dose (160 mg/Kg bwt) brought down the LPO up to 87% enhanced by 4-NQO. Among the enzymatic antioxidants, glutathione S-transferase (GST) was the most affected enzyme with 4-NQO and the least was catalase (CAT). Pretreatment with EAF (160 mg/Kg bwt), the restoration of antioxidants like glutathione peroxidase, superoxide dismutase, and CAT were found equal or less than 1.2 fold higher than that of the respective control values whereas, GST was observed to be the most restored antioxidant. Be reduced glutathione (GSH) and the least vitamin C over their control values. EAF restored the GSH and Vitamin E levels were found to be 1.2 fold higher than the respective control values.

The whole plant, Pergularia daemia (Family: Asclepediaceae), was extracted with 50% alcohol and a fresh batch of the plant material was successively extracted with petroleum ether, ethyl acetate and n-butanol to determine its diuretic activity. The diuretic activity of the different extracts at a dose of 400 mg/Kg was assessed orally in rats with furosemide as a standard drug using Lipschitzs test. All extracts except the petroleum ether extract showed significant increase (p < 0.001) in urine output. Urinary electrolyte excretion was also affected by the extracts: the alcoholic, ethyl acetate and n-butanol extract caused an increase in the urinary excretion of sodium and potassium ions. These findings suggest that among the mentioned extracts, ethanolic has the maximum diuretic activity followed by n-butanol extract.

The essential oils of branchlets and fruits of Juniperus excelsa subsp. excelsa and Juniperus excelsa subsp. polycarpos were examined for their antioxidant activity. The compositions of the essential oils were studied by GC and GC-MS. To evaluation the antioxidants activity of the volatile oils, pure components and positive controls at different concentrations, thin-layer chromatography (TLC) screening methods, diphenylpicrylhydrazyl (DPPH) assay, deoxyribose degradation test and modified deoxyribose degradation test were employed. The results of the present study demonstrate some antioxidant activity for the tested essential oils obtained from various parts of both plants. It indicates that the use of these essential oils, in very low concentrations, may be useful as a natural preservative. However before any final conclusion, it is suggested that the antioxidant activity of these oils should also be evaluated by using lipid solvent system methods.

Increased serum uric acid is known to be a major risk related to the development of several oxidative stress diseases. The aim of this study was to investigate the effect of parsley, quercetin and kaempferol on serum uric acid levels, liver xanthine oxidoreductase activity and two non-invasive biomarkers of oxidative stress (total antioxidant capacity and malondialdehyde concentration) in normal and oxonate-induced hyperuricemic rats. A total of 60 male Wistar rats were randomly divided into ten equal groups; including 5 normal groups (vehicle, parsley, quercetin, kaempferol and allopurinol) and 5 hyperuricemic groups (vehicle, parsley, quercetin, kaempferol and allopurinol). Parsley (5 g/Kg), quercetin (5 mg/Kg), kaempferol (5 mg/Kg) and allopurinol (5 mg/Kg) were administrated to the corresponding groups by oral gavage once a day for 2 weeks. The results showed that parsley and its flavonol did not cause any significant reduction in the serum uric acid levels in normal rats, but significantly reduced the serum uric acid levels of hyperuricemic rats in a time-dependent manner. All treatments significantly inhibited liver xanthine oxidoreductase activity. Parsley, kaempferol and quercetin treatment led also to a significant increase in total antioxidant capacity and decrease in malondialdehyde concentration in hyperuricemic rats. Although the hypouricemic effect of allopurinol was much higher than that of parsley and its flavonol constituents, it could not significantly change oxidative stress biomarkers. These features of parsley and its flavonols make them as a possible alternative for allopurinol, or at least in combination therapy to minimize the side effects of allopurinol to treat hyperuricemia and oxidative stress diseases.

Pistacia vera L., a member of Anacardiaceae family, has been used for sedation and analgesia in traditional medicine. In this study, the antinociceptive and anti-inflammatory effects as well as acute toxicity of the aqueous and ethanolic extracts of P. vera leaves were investigated in mice. The antinociceptive activity was studied using hot plate and writhing tests. The effect of the extracts against acute inflammation was determined using xylene-induced ear edema and the activity of the extracts, against chronic inflammation, was assessed using the cotton pellet test. The LD50 values of the infusion and maceration extracts were 0.8 g/Kg and 0.79 g/Kg, respectively. The aqueous and ethanolic maceration extracts of the P. vera leaves at the doses of 0.4 g/Kg and 0.5 g/Kg (IP), respectively, showed antinociceptive effects. The pretreatment of naloxone (2 mg/Kg, SC) inhibited the activities of extracts in hot plate test, but naloxone at the same dose could not inhibit the antinociceptive activity in writhing test. The extracts also showed anti-inflammatory effects in acute and chronic anti-inflammatory tests. The ethanolic extract was as effective as diclofenac in both inflammatory tests. The aqueous and ethanolic extracts of P. vera leaves demonstrated central and peripheral antinociceptive activities dose-dependently and the central effect may be mediated by opioid system. The extracts also demonstrated anti-inflammatory effects against acute and chronic inflammation.

In this study, the total 80% of MeOH extract and also petroleum ether, CHCl3, EtOAc, n-BuOH, and the remaining MeOH fractions obtained by solvent-solvent fractionation of the whole flowering samples of Centaurea bruguierana (DC.) Hand.-Mzt. ssp. belangerana (DC.) Bornm. (Asteraceae), namely “Baad-Avard”, collected from Borazjan in Bushehr Province (Bushehr, Iran) were investigated for larvicidal activity against malaria vector, Anopheles stephensi Liston, according to WHO methods. The mortality rate of total extract and petroleum ether fraction in concentration of 40 ppm were 28% and 86% respectively and the other fractions were inactive. The probit regression analysis for the dose-response to petroleum ether fraction treatment of larvae exhibited the LC50 and LC90 values of 15.7 ppm and 48.3 ppm, respectively. As results showed, the larvicidal activity of the petroleum ether fraction would be due to the nonpolar compounds in the plant which further isolation and purification would obtain the more active compounds in lower concentrations useful for preparation of biological insecticides.

Pycnocycla spinosa Decne. ex Boiss. var. spinosa (Fam. Umbelliferae) is an essential oil-containing wild plant growing in central part of Iran. Hydroalcoholic extract of Pycnocycla spinosa has antispasmodic and antidiarrheal activity. The aim of this study was to further investigate antidiarrheal and small intestinal transit effect of P. spinosa extract for a comparison with loperamide and dicyclomine. Male mice fasted over night with free access to water, were treated with P. spinosa extract, loperamide, dicyclomine or vehicle (p.o.). After thirty min, castor oil was given orally to the animals. In separate groups, magnesium sulphate was given in the beginning and 30 min after the extract or drugs were administered. The onset and number of wet defecation on the absorbent paper was recorded for each animal for 2.5 h. In another group of mice, intestinal transit of charcoal meal after the administration of extract, loperamide or dicyclomine was determined and compared with the control group. P. spinosa extract sharply reduced castor oil and magnesium sulphate induced diarrhea. The extract, in dose of 1 mg/Kg, had antidiarrheal effect similar to loperamide (2 mg/Kg) and with dose of 0.5 mg/Kg, its antidiarrheal action was greater than that of dicyclomine (5 mg/Kg). Unlike dicyclomine, P. spinosa extract significantly reduced the small intestinal transit of charcoal meal. However, its inhibitory effect on intestinal transit was less than loperamide. This study shows that anti-diarrhea l effect of P. spinosa extract is similar to loperamide. The inhibition of intestinal propulsion is a most likely mechanism that may account for anti-diarrhea l activity of the extract.

Cardiovascular diseases with an incidence of approximately 50% are the main causes of death in most advanced countries and an increasing trend in the developing world as well. The World Health Organization estimates that 12 million people per year worldwide die from cardiovascular diseases. Cardiovascular diseases are becoming an increasing problem worldwide and hypercholesterolemia has been correlated for coronary heart diseases. Nearly all lipoproteins are involved in this process including cholesterol carried by very low density lipoproteins (VLDL), remnant lipoproteins and low density lipoproteins (LDL). Currently, available hypolipidemic drugs have been associated with the number of side effects. Herbal treatment for hyperlipidemia poses no side effects and is relatively cheap and locally available. In view of this, the present study was carried out to investigate the effect of Menthe piperita on serum lipid levels of albino rats. Mentha piperita aqueous extract (100 mg/Kg, 250 mg/Kg p.o. daily) was fed for 3 weeks on fructose-fed rats and the levels of glucose, cholesterol, triglycerides, very low density lipoprotein, low density lipoprotein, and atherogenic index was measured. Twenty-four male Sprague Dawley rats were divided into four groups (6 per group). The results of present study indicate that Mentha piperita had significant beneficial effects against fructose-induced hyperlipidemia and showed good antioxidant activity. The aqueous extract of the plant produced a significant decrease (p < 0.05) in elevated levels of glucose, cholesterol, triglycerides, very low density lipoprotein, low density lipoprotein and atherogenic index and also increased the high density lipoprotein cholesterol levels and HDL-ratio without affecting serum insulin levels in fructose-fed rats.

The management of postprandial hyperglycemia is an important strategy in the control of diabetes mellitus and complications associated with the disease, especially in the diabetes type 2. Therefore, inhibitors of carbohydrate hydrolyzing enzymes can be useful in the treatment of diabetes and medicinal plants can offer an attractive strategy for the purpose. Vaccinium arctostaphylos leaves are considered useful for the treatment of diabetes mellitus in some countries. In our research for antidiabetic compounds from natural sources, we found that the methanol extract of the leaves of V. arctostaphylos displayed a potent inhibitory activity on pancreatic α-amylase activity (IC50 = 0.53 (0.53 – 0.54) mg/mL). The bioassay-guided fractionation of the extract resulted in the isolation of quercetin as an active α-amylase inhibitor. Quercetin showed a dose-dependent inhibitory effect with IC50 value 0.17 (0.16 – 0.17) mM.

The drug pump protein MRP2 is a membrane drug efflux transporter widely distributed in normal and tumor tissues. Its role is thought to be crucial for the disposition of many drugs and their substrates in different tissues. In this study, we aimed to examine the effects of systematic inflammation induced by lipopolysaccharide (LPS) on the expression and function of this transporter in rats. Jugular cannulated rats were injected intraperitoneally with LPS. Control rats received equal volume of sterile saline buffer. Rat liver MRP2 expression was analyzed at the level of mRNA through reverse transcription polymerase chain reaction. At various time points following drug administration (15 to 360 min), 250 µL blood samples were obtained from the cannula. The plasma was separated by centrifugation and stored at -20°C until HPLC analysis. Administration of LPS resulted in a slight but not significant increase in MRP2 mRNA levels 24 h after the treatment. In HPLC analysis, a rapid decrease in plasma concentrations of the MRP2 substrate (APAP) was observed at the initial time points. At later time points, the slope of the substrate concentration reached a plateau and paralleled to those of controls. It was postulated that due to the presence of the possible compensatory transport mechanisms in liver and non-hepatic tissues, changes performed to the pump activity were not completely in parallel to the expression of the drug efflux pump.

Nitric oxide (NO) is thought to be involved in spatial learning and memory in several brain areas such as hippocampus. This study examined the effects of post-training intrahippocampal microinjections of 1400W as a selective iNOS inhibitor on spatial memory, in anesthetized and non-anesthetized situations in rats. In the present work, 4-day training trials of animals were conducted. Spatial memory was tested 48 hours after the drug infusions. For microinjection of 1400W into CA1 region of the hippocampus in conscious animals, guide cannula was implanted into the CA1 area and 1400W was infused after recovery from surgical anesthesia. In anesthetized animals, 1400W was microinjected directly into CA1 region by Hamilton syringe during anesthesia. After completion of training, 1400W (10, 50 and 100 µM/side) were microinjected bilaterally (1 µL/side) and testing trials were performed 48 h after drug infusions in both groups of cannulated and non-cannulated rats. Significant reduction was observed in escape latency and traveled distance in animals that received 1400W (100 µM/side, *p < 0.05) via cannula after recovery in comparison with control group. Also, microinjection of 1400W (100 µM/side) in post recovery phase caused a significant (***p < 0.001) reduction in time and distance of finding the hidden platform in comparison with anesthetized situation. These findings suggest that 1400W has a significant improvement on spatial memory and memory enhancement induced by iNOS inhibitor can be affected by anesthesia in a period of time.

Enamel matrix derivative Emdogain (EMD) is widely used in periodontal treatment in spite of the fact that its effect on the developing embryo has not been elucidated. The aim of this study was to investigate the teratogenic effect of EMD on the rat embryo neural crest cells. The neural crest is a unique population of cells that migrates from the dorsal neural tube along defined pathways and produces various cell types including the melanocytes, neuronal and glial cells of the sensory, autonomic and enteric nervous system as well as the chromaffin cells of the adrenal gland. These cells have been used extensively for in-vitro studies of neurogenesis. Cultured cells by micromass culture method derived from midbrain of six embryos (13 day postcoitum; 34-36 smites) and exposed to various concentrations of EMD for 5 days at 37°C and differentiated foci were counted. Retinoic Acid (20 µg/mL) was used as standard positive control. These cells were stained using Mayer’s hematoxylin which is specific for staining differentiated cell nucleus. Neutral red staining determines cell viability rather than related cell differentiation but is used for normalization of Mayer’s hematoxylin results. At the concentration as low as 8 µg/mL of EMD, no toxic effect on fetal cells was observed and it is suggested that EMD has no teratogenic effect at studied concentrations.

In this research, we investigated the cytotoxic mechanisms of one of the widely used pharmaceuticals that are regularly associated with the adverse effects on the liver, sometimes leading to acute liver failure, diclofenac. Diclofenac liver cytotoxicity was associated with reactive oxygen species (ROS) formation and lipid peroxidation which were inhibited by antioxidants and ROS scavengers, ferric chelator, inhibitors of reduced CYP2E1 and CYP2C9, mitochondrial permeability transition (MPT) pore sealing agents and endocytosis inhibitors. Incubation of hepatocytes with diclofenac caused rapid hepatocyte glutathione (GSH) depletion which is another marker of cellular oxidative stress. Most of the diclofenac-induced GSH depletion could be attributed to the expulsion of GSSG. Diclofenac cytotoxicity was also associated with mitochondrial injury, lysosomal membrane rupture and release of digestive proteases which were prevented by antioxidants, MPT pore sealing agents, lysosomotropic agents and inhibitors of cytochrome P450 isoenzymes. These events could cause cytochrome C release from the mitochondrial intramembrane space to cytosol. The cytochrome C release could trigger activation of caspase-3 and apoptosis. We finally concluded that diclofenac hepatotoxicity is a result of metabolic activation by CYP2E1 and CYP2C9 and ROS formation, leading to a mitochondrial/lysosomal toxic cross-talk in the liver hepatocytes.

Pyridostigmine bromide (PB) is a reversible cholinesterase inhibitor. The aim of this study was to determine the effect of orally administration of single dose sustained-released tablet of pyridostigmine bromide (PBSR) on the frequency domain indices of heart rate variability (HRV). Thirty-two healthy young men were participated in this study. They were divided into 2 groups; the pyridostigmine group (n = 22) and the placebo group (n = 10). Electrocardiogram (ECG) was recorded at 10, 30, 60, 90, 120, 150, 180, 210, 240, 300 and 420 min after PBSR administration. At each time, simultaneously, a blood sample was prepared and PB plasma concentration was measured by high-performance liquid chromatography (HPLC) method. Statistical analysis showed that in different indices of HRV, there is a significant increase in low frequency (LF) band at 300 min, but no difference in high frequency band (HF). It also showed significant decreases in normalized high frequency band (Hfnu), normalized low frequency band (Lfnu) and LF/HF ratio at 120, 240 and 300 min after PBSR administration. Maximum plasma concentration of PB was 150 min after the administration. In conclusion, administration of a single dose PBSR can enhance the frequency domains indices of HRV and improve­­ sympathovagal ­balance­­­­.

Phytohemagglutinin (PHA) is a lectin, obtained from the red kidney bean that binds to the membranes of T-cells and stimulates metabolic activity, cell division, etc. The object of this research was the comparison between self made PHA (Indigenous) and imported commercial one, following conventional and High Resolution Cell Synchronization technique (HRCS). From each blood sample of healthy individual donor replicate cell culture with two different PHA (self-made and commercial imported) with same concentration were cultured simultaneously. For culture cells, 3-5 × 1066 cells were cultured in 4 mL medium(RPMI 1640 supplemented with 15 per cent heat inactivated fetal bovine serum, 0.1 mL Phytohemagglutinin was added and kept at 37°C in an atmosphere containing 5% CO2. The processing of mitotic division from 48 h and 72 h cultures was performed according to the standard and High Resolution Cell Synchronization technique. Cytogenetic studies were performed in 100 normal healthy blood donor individuals. Statistical analysis was performed by SPSS (version 16, Inc.USA) software.Our results indicate that the preparation of fresh Phytohemagglutinin at the time of cell division and cell culture procedure reveals satisfactory score. The overall frequency of mitotic index in our study was better when compared with commercial imported Phytohemagglutinin (p < 0.001).The significant differences in the results may be due to fresh preparation. However, cost effective, easy and nearest approach of this indigenous product and high demand for this product among health care services can be considered.

Amikacin has been shown to irreversibly suppressCochlear activity.The aim of this study is to assess the incidence of amikacinototoxicity in multidrug-resistant tuberculosis patients and riskfactors associated withthis ototoxicity.In this cross-sectional study, 41 patientswith multidrug-resistant tuberculosis (MDR-TB) were included.All patients received fixed dose of intravenous amikacin(500 mg/day) and anti-TB medications for six months. Baseline Pure-Tone Audiometry (PTA) was performed on all patients,before and during the drug treatment with the frequency range between 250 Hz and 8000 Hz. Patients were closely observed for the occurrence of symptomatic ototoxicity using a questionnaire. To find an association between the incidence of cochlear damage and patients’ demographics, all patients’ data were recorded. A total of 29 patients suffered from hearing loss (70.1%) (Male: n = 18; Female: n = 20).Using logistic regression, the incidence ofamikacinototoxicity was higher in men than in women. There was a negative correlation between the duration of the amikacintreatment and the difference in hearing thresholds(r = -0.34, p = 0.03). The mean of hearing threshold was significantly increased before and after theamikacintreatment((23.68 ± 19.26 vs. 38.93 ± 22.80) (p < 0.0001)). The incidence of hearing loss was remarkable in MDR-TB patients treating with amikacin. However, risk factors’ determination and monitoring of audiometric result variations could haveinfluenced the incidence of the amikacin ototoxicity.

New-onset hyperglycemia in patients admitted to intensive care units increases the risk of morbidity and mortality. Insulin resistance is frequently seen in the treatment of stress-induced hyperglycemia. Metformin, an oral anti-hyperglycemic agent, may introduce a new treatment protocol in critically ill patients with insulin-resistance hyperglycemia. Fifty-one non-diabetic traumatized patients with blood sugar (BS) levels more than 130 mg/dLwere introducedto three days of treatment with intensive insulin (50 IU) or metformin (1000 mg, twice daily) therapy. Clinical evaluationsincluded acute physiological and chronic health evaluation (APACHE II) and Glasgow Coma Scale (GCS). Experimental tests included BS level, mean arterial pressure (MAP), pH, HCO3, and lactate. Eight patients were excluded and 21 of remained patients treated with insulin and 23 with metformin. There was no significant difference in terms of the evaluated factors between the two groups at the time of admission. Although desirable BS level (BS < 130 mg/dL) was reached by three days of metformin treatment (p < 0.01),there was no significant difference in BS, MAP, pH and HCO3of insulin treated groupin comparison with metformin treated patients. The findings weresimilar for APACHE II and GCS as well. Although obvious studies are required, these findings may lead to effective therapies against stress-induced hyperglycemia.

Prescriptions written by general practitioners and medical specialists were studied and compared to determine the type, time of onset and clinical importance of drug-drug interactions (DDIs) in an attempt to reduce further complications. In 2007, 28, 956, 638 prescriptions and 15, 610, 912 prescriptions in 2008 were filled by pharmacies affiliated with medical science universities. These prescriptions, prescribed by physicians from 33 Iranian medical universities nationwide were then evaluated with a prescription processing software named Pardazesh Nosakh. After processing and analyzing the data, DDIs were discovered in 14 different medical specialists consisting of internists, cardiologists, neurologists, psychiatrists, neurosurgeons, general surgeons, infectious diseases, urologists, dermatologists, ENT, ophthalmologists, orthopedists, and pediatrician. The results were then analyzed through methods applied in the book of Drug Interaction Facts. The results revealed that in 2007-2008, 0.77% of prescriptions had DDIs out of which 0.67% were with significant clinical importance. The percentage of interactions with significant clinical importance was higher in prescriptions of medical specialists and of those, cardiologists and internists ranked top on the list, while dermatologists ranked the lowest. The most common interacting combination prescribed was digoxin and furosmide in 2007-2008, and captopril and triamteren in 2008-2009. Moreover, this study showed that polypharmacy was an important factor which led to DDIs. Drug interactions were common among outpatients prescribed multiple medications and the rate of DDIs increased with the number of drugs prescribed. It is our opinion that by being up-to-date on drug information and participating in related educational classes and workshops, physicians can increase the chances of choosing the correct drug treatment and hence significantly decrease possible DDIs side effects.

The purpose of this study was to determine the adherence to oral hypoglycemic medications and associated factors in type 2 Diabetes Mellitus patients who were referred to the Isfahan Endocrinology and Metabolism Research Centre (IEMRC). Convenience sampling was used to enroll 248 patients with type 2 diabetes in a prospective study at IEMRC from January 2007 to January 2008. Patients had to be on a stable dose of oral hypoglycemic medications (glyburide and metformin) or for 3 months prior to the study and willing to participate in consultation sessions with a pharmacist. Pill count and self report methods were used to measure the adherence. Mean (SD) of patients studied was 56.6 (8.9) years and 62% were females. The mean (SD) duration of diabetes in the study patients was 10.8 (6.1) years and 81.9% of them were literate with basic education. Non-adherence rates to metformin and glyburide were recorded in 39.7% and 35.3% of the study population respectively. Lower HbA1C levels and higher education were associated with higher adherence rates. Forgetfulness, confusion, fasting, adverse effects, complexity of medication regimen and disruption of routines were most commonly reported causes of non adherence. Prevalence of adherence to these two medications did not differ significantly between pill count (62.3%) and self report (62.8%), (p >0.05). Adherence rates did not vary by pill count and self report significantly. It was concluded that good adherence to medications was associated with a lower HbA1C profile; so it seems that pill count is a useful method in the clinical practice to identify non-adherent patients. Further studies are needed to find out efficient interventions to improve the patient’s adherence.

Different symptoms in Climacteric period, includes hot flash. Hormone replacement therapy (HRT) is common therapy for relief of menopausal symptoms but has possible contraindications and side effects. Recently Piascledine (combination of Avocado oil with Soybean oil) showed effects in reducing hot flash severity. Present study designed to compare the effects of HRT with Piascledine in treatment of hot flash. The cases of this study were sixty-six women at the age range of 40 to 70 years and complaints of menopause-induced hot flashing, whose last menstruation dated at least 6 months prior to the beginning of the study. The patients in this open label clinical trial, randomized to receive Piascledine capsule 1 mg or HRT (0.625 mg oral daily Conjugated Estrogen tablets, plus 2.5 mg continuous oral daily Medroxyprogesterone Acetate tablets) for 2 month. Hot flash property and severity was assessed via a daily check list and Visual analog scale. Climacteric symptom was measured before and after intervention using Greene Climacteric Scale (GCS) and Blatt-kupperman Menopausal Index (BKMI). Thirty-three eligible patients were allocated in each group. From the Piascledine group, one patient and from the HRT group, 16 patients weren›t willing to attend the study; therefore, 32 and 17 woman received treatment in Piascledine and HRT groups. 4 patients were withdrawn for vaginal bleeding and one for breast tenderness from HTR group. Hot flash severity in both groups decreased during the time similarly. With regard to GCS (p = 0.571) and BMKI (p = 0.891), the outcome was similar among the two groups. Due to low HRT compliance and its possible risks in long period of time and considering the same activity of soybean supplement and HRT in relieving the hot flash as menopausal symptoms in women, it seems that soybean supplements can be an alternative therapy to hormone.

Vitamin E is a potent reactive oxygen metabolites (ROM) scavenger. It is a lipid-soluble vitamin and its main function is to protect polyunsaturated fatty acids against oxidative stress. Twenty-five mechanically ventilated Intensive Care Unit (ICU) adult patients participated in a prospective randomized clinical trial receiving either placebo (10 patients) or 3 IM doses (1000 IU each) of vitamin E (15 patients). We determined plasma and bronchoalveolar lavage (BAL) fluid concentrations of vitamin E and superoxide dismutase (SOD). Among these 25 patients, there were 14 men and 11 women, aged 63.16 ±15.48 years (mean ± SD; range = 33 to 87 years). Vitamin E supplementation resulted in significant differences in plasma and BAL vitamin E concentrations between the two groups (p-value = 0.01, 0.01), decrease in SOD activities (not differ significantly in plasma (p-value = 0.23)), but with significant differences in BAL (p-value = 0.016) and progressive reduction in Acute Physiology and Chronic Health Evaluation II (APACHE II) (p-value = 0.52) and Sequential Organ Failure Assessment (SOFA) (p-value = 0.008) score in vitamin E group. From the results of this study, it seems that supplementation of vitamin E as a potent antioxidant, along with other supportive measures, can be beneficial in decreasing SOD total activity, ROM production and risk of organ failure in critically ill patients.