Iranian Journal of Basic Medical Sciences
Volume:12 Issue: 2, Summer 2009

7-prenyloxycoumarins including 7-isopentenyloxycoumarin, auraptene and umbelliprenin, and herniarin have been widely recognized as bioactive coumarins. This paper presents the ways to synthesis these compounds.7-prenyloxycoumarins were synthesized by reaction between 7-hydroxycoumarin (1 M) and relevant prenyl bromides (1.5 M) in acetone at room temperature. The reaction was carried out in the presence of DBU (1, 8-diazabicyclo [5.4.0] undec-7-ene) (2 M). After 24 hr, the mixture was concentrated under reduced pressure. The compounds were purified by column chromatography.Three bioactive 7-prenyloxycoumarins, namely, umbelliprenin, auraptene and 7-isopentenyloxycoumarin, together with herniarin were synthesized from 7-hydroxycoumarin under alkaline conditions (DBU) and then purified by column chromatography. The structures of the products were characterized by NMR spectroscopic method including 1H- and 13C-NMR experiments. The method of synthesis for 7-prenyloxycoumarins and herniarin which is presented here has not been reported yet. Moreover, for the first time, umbelliprenin was chemically prepared in this work.

Mesenchymal stem cells (MSCs) from large animals as goat which is genetically more closely related to human have rarely been gained attentions. The present study tried to isolate and characterize MSCs from goat bone marrow. Fibroblastic cells appeared in goat marrow cell culture were expanded through several subcultures. Passaged-3 cells were then differentiated among the osteogenic, adipogenic and chondrogenic cell lineages to determine their MSC nature. Differentiations were determined by RT-PCR analysis of related gene expression. To identify the best culture conditions for propagation, passage-3 cells were plated either at varying cell densities or different fetal bovine serum (FBS) concentrations for a week, at the end of which the cultures were statistically compared with respect to the cell proliferation. In this study, we also determined goat MSC population doubling time (PDT) as the index of their in vitro expansion rate. Passage-3 fibroblastic cells tended to differentiate into skeletal cell lineages. This was evident in both specific staining as well as the specific gene expression profile. Moreover, there appeared to be more expansion when the cultures were initiated at 100 cells/cm2 in a medium supplemented with 15% FBS. A relatively short PDT (24.94±2.67 hr) was a reflection of the goat MSC rapid rate of expansion. Taken together, fibroblastic cells developed at goat marrow cell culture are able to differentiate into skeletal cell lineages. They undergo extensive proliferation when being plated at low cell density in 15% FBS concentration.

Air pouch is a well-established inflammatory model in which fluid extravasations; leukocyte migration, angiogenesis and other parameters involved in the inflammatory response can be measured. In this study, the influence of gender on inflammatory parameters has been examined in the air pouch model. To induce air pouch, adult male and female Wistar rats were anesthetized, then 20 ml and 10 ml of sterile air were injected subcutaneously on the back on day 0 and day 3, respectively. On day 6, inflammation was induced by injection of 1 ml of carrageenan 1% into pouches. After 6 and 72 hr, the rats were sacrificed, pouch fluid was collected in order to determine exudates volume and the accumulated cells were counted using a hemocytometer. Pouches were dissected out and weighed. Angiogenesis of granulomatous tissue was assayed using hemoglobin kit. Analysis of our data demonstrated a sexually dimorphic pattern in inflammation parameters both in acute and chronic forms (P<0.05). The value of angiogenesis in the air pouch model in male rat was higher than that female rats (P<0.001). The degree of inflammation and angiogenesis induced in Wistar rat air pouch model is gender-dependent, suggesting that gender may be a key consideration in the design of inflammation experiments.

The fractions of Candida albicans have been used as an immunomodulator. The present work assessed the effect of different fractions of C. albicans on nitric oxide (NO) production by mice peritoneal macrophages. Cell wall and cytoplasmic fractions of C. albicans ATCC 10321 strain were extracted. Mice peritoneal macrophages were purified and cultured. Different concentrations of both fractions and also killed C. albicans cells were used for macrophages stimulation and evaluation of NO production. NO amount was detected in culture supernatants of macrophages by Griess reagent. Also, MTT assay was performed to assess the viability of macrophages.The results elucidated that suppressive effect of cell wall proteins on NO release was significant at the dose of 100 μg/ml (P=0.01), while cytoplasmic fraction increased NO amount at the dose of 1 μg/ml compared to the control group (P=0.003). Augmentation of NO production was statistically significant at 200 killed C. albicans per well (P=0.006).According to our findings, cytoplasmic fractions and killed C. albicans have a positive effect on NO production by peritoneal macrophages, while cell wall fractions did not. Therefore, it is proposed that C. albicans fractions can be studied more as inflammation modulators.

Comparison of prooxidant-antioxidant balance (PAB) assay with crocin assay.Twenty eight serum samples were chosen, PAB and the total antioxidant capacity were measured by PAB assay and crocin, respectively, and the correlation of both assays, along with their correlation with other clinical and biochemical parameters, were determined. A significant negative correlation was established between PAB assay and crocin assay. Also a significant negative correlation was established between PAB and uric acid and creatinine.The results showed that by increasing the total antioxidant capacity, which is showed by crocin, the PAB shifts in favor of antioxidants, which is showed by PAB assay. Now, it could be considered that the PAB, along with other risk factors, might help in the prediction of the risk for cardiovascular events; and further research could clarify whether by application of PAB assay and appropriate interventions for correcting oxidative stress, progression of the cardiovascular disease could be reduced.

Although, type 2 diabetes is the most frequent type of diabetes, its main cause is yet to be clarified. Several environmental and genetic parameters are believed to be involved in diabetes. It has also been established that cytokines play key roles in pathogenesis of diabetes. Expression of cytokines is different from person to person and in different societies. Several studies showed that polymorphisms of +874 of interferon-gamma (IFN-g) and -590 of interleukin-4 (IL-4) are associated with the regulation of expression of these genes. This study was aimed to find polymorphisms of these regions in type 2 diabetes patients.In this experimental study peripheral blood samples were collected from 160 type 2 diabetic patients and 160 healthy controls. DNA was extracted by salting out method. Polymorphisms of +874 of IFN-g and -590 of IL-4 were analyzed by ARMS-PCR and RFLP-PCR.Our findings indicated that TT genotype of IFN-g was increased in type 2 diabetic patients compare d to the control but difference was not significant.Our results didn’t show any significant difference between IL-4 genotype in diabetic and healthy controls either.Our results suggested that TT genotype of IFN-g can be associated with diabetes. This association can be described by the fact that over e xpression of IFN-g shift s immune system to Th1; therefore, pancreatic cells can be miscarried by immune cells.

Due to increasing emergence of drug-resistance in Helicobacter pylori isolates, traditional plants are potentially valuable sources of novel anti-H. pylori agents. In this research, anti-H. pylori activity of the organic extracts of twenty native Iranian plants was determined against ten clinical isolates of H. pylori. Disc diffusion was used to determine the biological activity of 20 plant extracts as well as 8 antibiotics commonly used to treat H. pylori infections. Minimum inhibitory concentrations were also measured by tube and agar dilution methods for the biologically active plant extracts.Of the twenty plant extracts analyzed, sixteen exhibited good anti-H. pylori activity, using disc diffusion. The ten most active extracts were Carum bulbocastanum, Carum carvi, Mentha longifolia, Saliva limbata, Saliva sclarea, Ziziphora clinopodioides, Thymus caramanicus, Glycyrrhiza glabra, Xanthium brasilicum andTrachyspermum copticum. Minimum inhibitory concentrations measured for the 10 biologically active plant extracts were within the range of 31.25 to 500 mg/ml. Among the ten plant extracts effective against H. pylori clinical isolates, Carum carvi, Xanthium brasilicum and Trachyspermum copticum showed the highest activity.

Quinazolinones are heterocyclic compounds, with biological and pharmacological activities, such as inhibiting some proteins, enzymes and reducing blood lipids. Following previous results of our group, effects of two new derivatives of quinazolinones 9(3)-quinazolinone-2-propyl-2-phenylethyl (QPPE) and 9(3)-quinazolinone-2-ethyl-2–phenylethyl (QEPE) on livers, intestines and kidneys of newborn Balb/C mice were investigated. Pregnant mice were divided into four groups of control, sham, experimental 1, treated with QPPE, and experimental 2, treated with QEPE. Experimental groups received 100 mg/kg body weight (most effective dose) of QPPE and QEPE, sham groups received methyl cellulose 0.05% (the solvent) and control groups received distilled water, intraperitoneally (IP), on day 8 of gestation. Five days after birth, livers, intestines and kidneys were removed, fixed in formalin 10%, stained with hematoxylene and eosin for histological and pathological studies.Results showed appearance of fatty changes in livers, an increase in diameters of hepatocytes and central veins of livers, and reduction in the lengths of villi of proximal, middle and distal segments of newborn Balb/C mice intestines. Furthermore, there was a diminished diameter of the lumen of the proximal tubules, and average diameter of the lumen of distal tubules which led to an increase in the number of glomeruli cells of newborn Balb/C mice kidneys.Regarding inflammation in different parts of the kidneys, livers and intestines, our investigations suggest that quinazolinones may have some toxic effects on embryos.

This study planned to assess the value of PCR IS6110 assay in tissue specimens of needle pleural biopsy in patients suspicious to pleural tuberculosis.Sixty eight patients with lymphocytic exudative pleural effusion underwent pleural biopsy. Tissue samples were sent for pathologic examination and PCR IS6110 assay. The results of PCR reported as positive/ negative and assessed according to the current gold standard pathologic diagnosis.Twenty nine patients had tuberculous and 12 had malignant pleural involvement, respectively. The remaining 27 samples were reported as non-specific pleurisy. Results of PCR were positive in 35 out of 68 total subjects and in 19 out of 29 TB patients. Sensitivity and specificity of PCR were calculated as 67.9% and 62.5%, respectively. An acceptable sensitivity and specificity for PCR examination of pleural tissue can serves it as a useful adjunct in undergoing needle pleural biopsy for possibility of tuberculosis.