Iranian Journal of Biotechnology
Volume:7 Issue: 3, Summer 2009

Given the difficult accessibility of articular cartilage as chondrocyte source for fabricating cartilage construct, alternative chondrocyte source needs to be considered. The present study examined the chondrocytes from costal cartilage for their suitability as cellular material for cartilage tissue engineering. For this purpose, about 5×105 passaged-5 chondrocytes, isolated from rat knee and rib hyaline cartilage, were separately mixed with one ml alginate solution; the mixture was dropped in calcium chloride solution as 2-mm-beads and incubated for a period of 2 months during which structural features, proliferation rate, and expressional level of some gene were determined by microscopy, MTT test and real-time PCR analysis respectively. According to our results, both articular and costal chondrocytes were observed as organelle-rich round to oval cells embedded in lacuna-like cavity in alginate. The propagation pattern of the cells in alginate was the same; undergoing proliferation during the first two-third and ceasing mitosis during the last third of culture period. The level of aggrecan and collagen II (cartilage-specific) gene expression at costal cultures were comparable to those of articular cultures; the expression levels tended to be very low at early days and largely up regulated at the end of cultivation period (day 60). Interestingly, in contrast to the culture of articular cells, the level of collagen I (bone-specific gene) expression was in negligible amount at costal culture (P<0.05). Taken together, our data suggests that the costal chondrocytes could be considered as appropriate cells for creating 3D cartilage constructs for cartilage defects. Keywords: Costal and articular chondrocytes, proliferation, gene expression, structure, alginate.

In the present study، genotyping of short tandem repeat (STR) loci including CSF1PO، D16S539، F13A01، F13B، LPL and HPRTB was performed on genomic DNA from 127 unrelated individuals from the Iranian province of Isfahan. The results indicated that the allele and genotype distributions were in accordance with Hardy-Weinberg expectations. The observed heterozygosity (Ho)، expected heterozygosity (He) as well as forensic and paternity indices including power of discrimination (PD) and exclusion (PE)، polymorphism information content (PIC)، typical paternity index (PItypical) and probability of paternity (W) were determined for the examined STR alleles. Also، genetic diversity index (GD) and population parameter (θ) were calculated in the six loci. The combined power of discrimination (Pdcombined) and combined probability of exclusion (PEtypical) were 0. 9999998 and 0. 999856 over the six loci، respectively. Together، the examined STR loci in this study have proven a relatively high genetic variation in Iranian population. The data could be used in construction of a genetic database for this population.

Present investigation is directed to biotransform, an anti-inflammatory compound, meloxicam by employing five actinomycetes cultures with the aim of producing new bio-active derivatives. Among the cultures screened, Streptomyces griseus NCIM 2622 was found to oxidise meloxicam into two metabolites whereas, S. griseus NCIM 2623 could oxidise meloxicam to only one metabolite in significant quantities. The metabolites formation in culture broth was monitored and confirmed by HPLC analysis. The structures were elucidated based on LC-MS-MS data and previous reports as 5-hydroxymethyl meloxicam and 5-carboxy meloxicam. From the above results it is concluded that S. griseus NCIM 2622 can be employed to oxidise meloxicam.

Abstract α1-antitrypsin (AAT) is a protease inhibitor that its primary function is to inhibit neutrophil elastase, a protease that can degrade all of the constituents of the pulmonary connective tissue matrix. AAT administration purified from blood is beneficial for AAT deficiency and cystic fibrosis and several other inflammatory diseases. A less expensive supply of AAT from recombinant producer can reduce patient treating cost compare to blood purified source. To facilitate and accelerate the production of eukaryotic proteins with correct post-translational modifications, we have developed a protein production system based on the transduction of CHO and COS-7 cells using lentivirally vectors. To accomplish this human AAT cDNA was cloned into a replication-defective lentiviral vector. The transgene AAT-Jred chimer was transferred to CHO and COS-7 cell line using this vector and its expression was visualized by fluorescent microscopy. The mRNA levels expression of AAT genes were determined using RT-PCR and its secretion from both cells into the medium was determined using ELISA.The results show that by employing a lentivirally vector efficient genetic loading of CHO and COS-7 cells with the AAT gene was achieved. The expression of the AAT gene in target cells was confirmed using RT-PCR. ELISA assay showed both CHO and COS-7 cells were active for secretion of AAT to medium.It has been concluded that by using a Lentivirus-based gene delivery system, we were able to express high amounts of recombinant human AAT protein in the CHO and COS-7 cell lines and this expression system possess key properties that ensure their employment to deliver therapeutic gene to mammalian cultured cells

Optimal control problems appear in several engineering fields and in particular on the fed-batch fermentation processes. Since Simulated Annealing Method has less been used in bioprocesses; we prefer to use this method of optimization in this research because of two reasons. First the uncertainty of the model of these processes and also these models depended all on a lot of parameters. The optimal feeding profiles of a fed batch process for three cases obtained by means of a stochastic algorithm. The accepted profiles were smooth and similar to those derived analytically in other studies. With these result effect of cell mass and glucose concentration was shown on feed profile and ethanol production. And ability of simulated annealing was tested for this kind of model; however the answer was so good.

In this paper computational design of novel 3- (4-chlorophenyl) -5- (3hydroxy-4-etoxyphenyl) -4،5-dihydro-1H-pyrazole-1-carboxamide is reported. The structure and relative energies of the target molecule are predicted using Hartree- Fock method. The methods of theoretical chemistry have been used to elucidate the molecular properties. The analysis of molecular descriptors defined by Lipinski has shown that the candidate drug obey ''rule of five''. The solubility of drug in water has been determined as it is of useful importance in the process of drug discovery and development from molecular design to pharmaceutical formulation and biopharmacy. Electrostatic potential maps have been constructed and the nature of Electrostatic Potential maps for the predicted bioactive conformations to identify the electrophilic and nucleophilic regions has been discussed. All toxicities associated with candidate drug have been calculated. P-glycoprotein (P-gp) is a cell membrane-associated protein that transports a variety of drug substrates. P-gp expressed in normal tissues as a determinant of drug pharmacokinetics and pharmacodynamics been examined. Drug plasma-protein binding and volume of distribution are one of the many factors which influences bioavailability of a drug، hence its value has also been calculated. To avoid rejection of drugs، it is becoming more important to determine pKa، absorption، polar surface area and other physiochemical properties associated with a drug، before synthetic work is undertaken.

The main objective of this study is to find out wheather trehalose affects photosynthesis genes (PS1-H and LHCB1) expression. At the first step، expression levels of 2 photosynthetic genes PSI-H and LHCB1 were determined in 10 d seedlings of WT growing on 100 mM sorbitol or trehalose using Q-PCR method. Results showed that PSI-H and LHCB1 transcript levels were reduced in the presence of 100 mM trehalose. Then cDNA of PS1-H and LHCB1 were PCR amplified and inserted into pGEMT easy vector as a universal vector and pBin19 (under control of 35S promoter) as a plant expression vectors. The results from molecular analysis and sequencing showed that PSI-H and LHCB1 genes have been cloned in correct orientation in both plasmids. Re-combinant plasmids were transferred to Arabidopsis seedlings via Agrobacterium tumefaciens mediated transformation method. Re-transformed lines showed different phenotypes compared to WT. Expression levels were determined in different re-transformed lines compare to WT seedlings. PS1-H and LHCB1 gene expression in re-transformed lines showed an induction with presence of inserted cDNA. A significant reduction in photosynthetic genes transcript abundance was observed by feeding of 100 mM trehalose، compare to 100 mM sorbitol. These findings showed that trehalose has inhibitory effect on photosynthetic genes expression.