Archives of Advances in Biosciences

Since the commercial introduction of insulin in 1923, thyroid hormone in 1934, factor VIII in 1948, and calcitonin in 1970, plenty of new proteins have been firmly established as therapeutic agents. Currently there is great interest in production of the protein-based drugs which are used for both preventative and therapeutic purposes. Generally, the pharmaceutical proteins perform the same function as naturally-occurring proteins in body. Traditionally animal and plant sources have been used to obtain protein-based drugs which are indeed expensive and available in the limited supply. For instance, the human growth hormone was taken from human corpses and insulin required to treat diabetes was collected from slaughtered pigs, cows and other animals. Nowadays, the use of recombinant DNA (rDNA) technology has enabled the production of large quantities of protein drugs as cost-efficient sources. The cloned genes are then genetically engineered into microorganisms or animals to produce the protein of interest. Production of protein-based drugs in rDNA technology can be achieved through the transgenic animals (pharming), microorganisms or through hybridomas. Of the more than 200 pharmaceutical proteins which have been investigated to date, more than half are undergoing research and development, about 100 are in clinical trials, and a dozen or so have already been marketed. The most important indications for them are cardiovascular disorders, tumors, autoimmune diseases, and infections. Overall, the protein-based drugs have found significant therapeutic potential, various clinical applications and growing market worldwide. On the other hand, several research centers/institutes in the field of life sciences have been established in our universities during the past decade. Now, these research centers and institutes could be pioneer for acquiring the big responsibility in different stages of production and commercialization of many protein-based drugs.

 Reza Yousefi

    The relation between single nucleotide polymorphisms (SNPs) and some diseases has been concerned by many researchers. Also the missing SNPs are quite common in genetic association studies. Hence, this article investigates the relation between existing SNPs in DNMT1 of human chromosome 19 with colorectal cancer. This article aims is to presents an imputation method for missing SNPs not at random. In this case-control study, 100 patients suffering from colorectal cancer consulting with the Research Institute for Gastroenterology and Liver Disease of Shahid Beheshti University of Medical Sciences were considered as the case group and 100 other patients consulting with the same research institute were considered as the control group and the genetic test was applied in order to identify the genotype of the 6 SNPs of the DNMT1 of chromosom 19 for all the patients under investigation. The obtained data were analyzed using logistic regression, then a fraction of the data was eliminated both at random and not at random and the imputation was done through the EM algorithm and the logistic regression coefficients variation before and after the imputation was compared. The results of this study implied that in both methods, at random and not at random missing SNPs, the estimation of the logistic regression coefficients after the imputation through EM algorithm has a greater correspondence to the results obtained from the complete data in comparison with the method of eliminating the missing values.

Down syndrome (DS), trisomy 21, is the most common chromosomal syndrome that affects one in 600-800 live births. The advanced maternal age is the only well known risk factor to cause DS. Our study revealed that many young mothers produced DS children than advanced age mothers in India. A total of 150 suspected DS cases were investigated cytogenetically. Randomly selected 200 healthy families in South India were used as controls. Logistic regression was performed on case-control dataset which was generated by randomly selecting the child from each of the control families. Pedigree analyses indicated that the maternal grandmothers had advanced age during conception of their daughters who gave birth to DS child. Case-control status was used as dependent variable, whereas parental and grandparental age was used as covariates. Logistic regression was reported as odds ratios, univariate and multivariate. The age of maternal grandmother showed highly significant difference in odds ratio, indicating that the advanced age of maternal grandmother was the possible risk factor.  Therefore, it is important to sort-out the effect of advanced age mothers vs grandmothers on increased frequency of DS reported in different populations.

Photodynamic Therapy (PDT) using 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) has been considered as a new method for treating neoplasms. However, ALA-PDT is suboptimal for thick tumors. Searching for new approaches, we investigated the effect of adding differentiation therapy (DT) with Vitamin E succinate (VES) to PDT in human prostate LN-CaP-FGC10 cancer cells in vitro. The purpose of DT was to reverse the lack of differentiation in cancer cells and to enhance the effectiveness of ALA- dependent PDT.

Three groups of cells were grown on RPMI1640 culture medium supplemented with 10% FBS. The cells included: ALA-PDT cells, which received 0.3 mM ALA for 4 hours at dark, and exposed to 532 nm, 50 mW Nd-YAG laser beam for 3 min; DT+ALA-PDT cells, which received 6 µg/mlVES for 24, 48 and 72 hours, followed by the addition of 0.3 mM ALA for 4 h and exposure to Nd-YAG laser beam for 3 min; control cells which were untreated. After 24 h, the percentage of cell viability was determined by MTT assay. Accumulation of PpIX was measured by spectrophotometry and fluorescent microscopy. Mechanism of induced cell death was investigated via Hoechst staining. The combination of both factors (VES and 5-ALA) lead to a significant increase in cell death after 72 h. Induction of differentiation augmented PpIX accumulation in cells treated with ALA. Elevated intracellular PpIX levels resulted in an enhanced lethal photodynamic sensitization of VES plus ALA-treated cells after 72 h. Apoptotic cell death by both ALA-PDT and VES-ALA-PDT was confirmed by Hoechst staining. Our data suggest that VES used in combination with 5-ALA may provide a new combinatorial approach for treating certain cancers.

     One of the most important aspects in recombinant biologic production, based on GMP rules, is the accuracy of final product quality control, especially assessment of host cell macromolecules contamination rate in final product. The purification requirement can be eliminated when the yeast cell containing the recombinant protein is used as a host cell. It is possibile that the final product contaminated  to the host cell protein during purification stages of HBsAg (HBV vaccine). The protein purification costs depend on the purification  procedures required. Nowadays several companies produce commercial kits for identification and assessment of host cell protein contamination based on ELISA and Western blotting methods. But high prices, difference in sensitivity and lack of easy access to these kits sometimes create problems. So, in this study, two methods of Ammonium sulphate and caprilic acid precipitation technique were used separately for IgG purification. The results showed that IgG purification increased by 97% in caprylic acid method, compared with only a 77% increase in ammonium sulphate method. There were also significant differences in specificity and sensitivity between our standardized ELISA technique and using commercial kit (Cygnus CHO HCP). 

      The aim of this study is to compare the sensitometric properties of commonly used radiographic films processed with chemical solutions in different workload hospitals. The effect of different processing conditions on induced densities on radiologic films was investigated. Two accessible double emulsions Fuji and Kodak films were exposed with 11-step wedge and processed with Champion and CPAC processing solutions. The mentioned films provided in both workloads centers, high and low. Our findings displays that the speed and contrast of Kodak film-screen in both work load (high and low) is higher than Fuji film-screen for both processing solutions. However there was significant differences (p=0.000 and 0.028) in films contrast for both workloads when CPAC solution had been used. The results showed base plus fog density for Kodak film was lower than Fuji. Generally Champion processing solution caused more speed and contrast for investigated films in different conditions and there was significant difference(p=0.01) in 95% confidence level between two used processing solutions. Low base plus fog density for Kodak films provide more visibility and accuracy and higher contrast results in using lower exposure factors to obtain better quality in resulting radiographs. In this study we found an economic advantages since Champion solution and Kodak film are used while it makes lower patient dose. Thus, in a radiologic facility any change in film processor/processing cycle or chemistry should be carefully investigated before radiological procedures of patients are acquired.

Esophagus cancer is the eighth most common cancer worldwide and particularly high in an area extending from the southern border of the Caspian Sea in Iran across central Asia to China. Since information about this mysterious disease is poor, proteomics may be solving this enigma. Altering gene expression in cancer cell is a remarkable indicator can be detected by proteomics techniques and bioinformatic analysis. In this study, normal and cancerous cells were obtain from patients, total proteins were purified by standard methods, and proteins separated by two dimensional electrophoresis (2DE). Some of proteins were identified by Mass spectrometry (MS-MALDI method). By using bioinformatic analysis illustrate molecular mechanism in this disease. Analysis of gels base on Flicker software and Mass Spectrometry led the same result. 61 protein spots detected in both gels that 21 spots have down regulated and 12 spots have up regulated in cancerous cell than normal. About 14 spots were disappeared in cancer cell while 14 new spots expressed. By using flicker detected 8 Protein that refer to TRFE, SZ07, C1 TC, Kininogen, anexin, keratin, fructosebisphosphate aldolase A and heat shock. Mass spectrometry (MS-MALDI method) identified anexin, keratin, fructosebisphosphate aldolase A and heat shock.  Identified proteins were functionally categorized based on Gene Ontology (GO) annotation terms using the DAVID program package. The major molecular functions that annotated with PIR include phosphoprotein, disease mutation while annotated by GO include response to organic substance, response to wounding  and cellular homeostasis. The cellular component and molecular function presenting the greatest enrichment that concluded two clusters that the two most importants are cellular homeostasis and extracellular region part. Results reveal that the most of molecular function in cancerous tissue maintenance cellular homeostasis, cell regeneration and repair, so tissues undergo stress try to survive. It can be also concluded that aldolase A, fructose-bisphosphate, keratin 14,  formyltetrahydrofolate synthetase and transferrin can be some diagnostic biomarkers and also drug targets in esophagus cancer.

Systemic lupus erythemathosus is an autoimmune disease that affected many various types of tissues in 10% of world population and over 30 genes has associated with it. Nouroimmunoendocrynology concepts have shown that immune system could be affected by neuron system and vice versa, 5-hydroxytryptamine receptor a (5HT3Ra) was studied as a main receptor in these relations.

In this study, peripheral blood sample were collected from (SLE) patient and normal individuals. The total cellular RNAs were extracted and the cDNAs were synthesized. This process was followed by real-time PCR using specific primers for 5HT3Ra gene and beta-actin gene as internal control. Eventually PCR products have been sequenced.

Results of this study suggested that this special receptor expressed in polymorpho-nuclear cells. We found over expression of 5HT3Ra in patients in comparison with healthy individuals group. Interestingly, some nucleotide changes have been found in 5HT3Ra gene in patients but not found sequential nucleotide changes in healthy individuals group.

This study supposed that over expression of 5HT3Ra gene in SLE patients lead to over activation of immune cells that derived from over stimulation  of them from serotonin blood serum that finally lead to autoimmune reactions that terminated in SLE.

    Endometriosis is a painful reproductive disease afflicting about up to 20% of women. It is one of the most frequent benign gynaecological diseases, however, little is known about the pathological of endometriosis. Over the past decade, high-throughput proteomics technologies have evolved considerably and have become increasingly more commonly applied to the investigation of female reproductive disease, including endometriosis. In this mini-review the authors look at the application of proteomics technologies in order to find biomarker associated with endometriosis.

The antidepressant effects of aloe vera hydro alcoholic extract at different concentrations were compared with the fluoxetine-treated and the control groups of mice using forced-swimming, FST and open box, OFT tests. The mice were evaluated in five groups (control, taking aloe vera at the dosage levels of 150 mg/kg, 300 mg/kg, and 450 mg/kg, and finally fluoxetine at a dose of 10 mg/kg) by the FST and OFT tests on 1^st, 7^th, and 14^th days. The results of the OFT test showed no significant differences between these five groups. The results of FST test indicate the antidepressant effects of aloe vera even at low doses and it was found that the effect of fluoxetine at a dose of 10 mg/kg was equivalent to the effect of aloe vera at a dose of 150 mg/kg for the reduction in immobility time in mice in FST test. According to the results obtained from FST test, the antidepressant effects on mice treated with the 450 mg/kg dose of aloe vera showed better recovery as compared with other groups on 1^st, 7^th, and 14^th days. With regard to the experiments performed at different times, all the evidence pointed to the conclusion that the antidepressant effect of aloe vera was more than the control group.  Based on the results of the OFT and FST tests, aloe vera extract at different doses, has favorable antidepressant effects on mice as compared to the fluoxetine-treated and the control groups  and the better effects were seen by increasing the dose and duration of drug use.