Background: Vibrio cholera toxin B (CTB) subunit is the pentameric non-toxic portion of cholera toxin (CT) which is responsible for the holotoxin binding to the GM1 ganglioside receptor present on nucleated cells. this study was to produce, purify, and verify recombinant CTB (rCTB) subunitin prokaryotic system.Materials and Methods: In this experimental study, rCTB expression vector (pET-28a) which could be induced in E. coli (BL21) was designed and synthesized. Then the recombinant expression strains containing the result of IPTG interaction were induced and the rCTB was generated on small and large scales. The rCTB produced through Ni2+-charged resin, after refolding and free of possible CTA contaminants, was extracted. After PURIFICATION, rCTB was verified by Western blotting.Results: The results indicated the level of PURIFICATION to be about 480µg of purified active pentameric rCTB for each liter of the induced culture. Also, Western blotting analysis showed that recombinant CTB is strongly and specifically recognized by polyclonal antibodies against the cholera toxin.Conclusion: The findings of this study demonstrated that E.coli is an available host for production of CTB. In addition, the designed host and vector can be used in large scale production of this protein.

Background: Inhibin is a protein synthesized by granulosa and sertoli cells which preferentially inhibit pituitary secretion of FSH through a negative - feedback. Studies have showed that measurement of inhibin has a clinical role in understanding the fertility status of men and women and could also be used as a prognostic marker for pre-eclampsia and Down syndrome in fetus. There are many different multi-step procedures for the PURIFICATION of inhibin. In this study we attempted to purify inhibin from human follicular fluid using immunoaffinity techniques.Materials and Methods: Follicular fluid (FF) collected from women referring to Avicenna Infertility Clinic, was filtrated and subsequently concentrated by ammonium sulfate. The presence of inhibin in follicular fluid was detected using enzyme-linked immunosorbent assay. In order to purify inhibin, an affinity chromatography column using anti-inhibin monoclonal antibody was prepared. The purified protein was analyzed through SDS-PAGE and western blotting after a protein solution load of human inhibin into the column.Results: The SDS-PAGE results of affinity proved the presence of inhibin following silver staining appeared as a single 32 kDa band. Western blotting revealed that specific antiinhibin antibody was able to recognize inhibin epitopes. The present protocol for PURIFICATION of inhibin is a technique with 92% yield.Conclusion: This method is a sensitive procedure for high yielded productions of inhibin compared to the previously described methods, using HPLC and gel filtration. However, since preparing and obtaining follicular fluid, as a main source of inhibin, is rather difficult but suitable for laboratories, recombinant inhibin is recommended for mass production.

Background and Objectives: Streptokinase is used clinically as an intravenous thrombolytic agent for the treatment of acute myocardial infarction and is commonly prepared from cultures of Streptococcus equisimilis strain H46A. The objective of the present study was the production of streptokinase from strain H46A and PURIFICATION by chemical reduction method.Materials and Methods: The rate of streptokinase production evaluated under the effect of changes on some fermentation factors. Moreover, due to the specific structure of streptokinase, a chemical reduction method employed for the PURIFICATION of streptokinase from the fermentation broth. The H46A strain of group C streptococcus, was grown in a fermentor. The proper pH adjusted with NaOH under glucose feeding in an optimum temperature. The supernatant of the fermentation product was sterilized by filtration and concentrated by ultrafiltration. The pH of the concentrate was adjusted, cooled, and precipitated by methanol. Protein solution was reduced with dithiothreitol (DTT). Impurities settled down by aldrithiol-2 and the biological activity of supernatant containing streptokinase was determined.Results: In the fed –batch culture, the rate of streptokinase production increased over two times as compared with the batch culture and the impurities were effectively separated from streptokinase by reduction method.Conclusion: Improvements in SK production are due to a decrease in lag phase period and increase in the growth rate of logarithmic phase. The methods of PURIFICATION often result in unacceptable losses of streptokinase, but the chemical reduction method give high yield of streptokinase and is easy to perform it.

In the fisheries industry, compared to other food industries, a large volume of byproducts such as bone, skin, and viscera are produced, which are costly to treat and dispose of in the environment. The rapid growth of tilapia and its marketability have also been considered. The value of proteins as the main source of amino acids in human health has been proven. In addition to their nutritional value, proteins have biological functions. These proteins show their functions through bioactive peptides. Production methods of bio peptides affect their biological activity. The articles showed that the use of enzymatic hydrolysis method for therapeutic applications makes them more stable than fermentation and solvent extraction methods and is a safer method than the other two methods. Bioactive peptides have physiological effects such as antihypertensive, antioxidant, antimicrobial, anti-cancer, anti-diabetic, etc. and have many applications in the food, pharmaceutical and cosmetic industries and can be added to the food industry. The purpose of this article is to review the extraction of bioactive peptides from fish waste.

The proper consumption of plant wastes could not only provide a possible source of natural products, but it also is an environmental friendly research. The aim of this study was to use grape wastes as a source of peroxidase. In practice, one isoenzyme of peroxidase (POD1) was partially purified from Vitis vinifera wastes, the plant which is widely harvested in Iran. The activity of this novel peroxidase was determined using guaiacol as its substrate. The new peroxidase was partially purified and its kinetic parameters determined. The values of Km and Vmax of peroxidase for guaiacol were 83. 2 mM and 0. 35 M/min respectively. Optimum pH and temperature were determined for guaiacol to be 6. 2 and 60 C respectively. According to SDS-PAGE results, the molecular weight of isozymes was 38-40 KD. The results indicate that agricultural leftovers from Vitis vinifera are a considerable source for a peroxidase with reliable kinetic behaviors.

BACKGROUND AND OBJECTIVE: Human- derived antibodies, which are prepared by expensive methods are used in diagnosis, prophylaxis, and treatment of diseases. The aim of this study was to develop an effective and inexpensive method for PURIFICATION of human serum immunoglobulins. METHODS: Human serum immunoglobulins were precipitated by ammonium sulfate (50% saturation) and then PURIFICATION was done by thiophilic chromatography (T-gel). PURIFICATION of antibodies was evaluated by cellulose acetate electrophoresis. Concentration of antibodies (IgG, IgM, IgA) in serum and the resulting pooled fractions were estimated by single radial immunodiffusion (SRID) and percentage of recovery was calculated. FINDINGS: Analysis of products by electrophoresis showed only one band in γ-region. Recovery of the purified IgG was 75% while that for IgM was considerably low (about 17%). CONCLUSION: The purified immunoglobulins were eluated by a low salt buffer closely neutral pH, thus post treatment of product was not necessary. Thiophilic chromatography causes the PURIFICATION of immunoglobulins at a stage after ammonium sulfate precipitation. The purified immunoglobulins in this study could be suitable for immunochemistry methods.

1. Prelude Surface water, especially rivers are the most important sources of water for drinking and agricultural purposes. The parameters involved in the ecosystem of the river covers a wide range and since the rate of elimination, some commonality, decrease and increase of pollutants and the factors influencing them is different, and there is in some cases, addressing all the features and parameters of water quality is not possible. In this regard, water quality parameters such as DO, BOD, COD, NH4, NO3 and PO4 in the Abbas Abad river of Hamedan were evaluated. The purpose of this study, was to simulate water quality parameters of Abbas Abad River with the QUAL2KW models and estimate the amount of pollution entering Abbas Abad River, without making environmental problems of rivers with the river enjoying self-PURIFICATION capacity. Also, it is noteworthy that no research has been done using this model in Abbas Abad river, so the results can be used in developing control programs pollutant load (TMDL) for Abbas Abad river, and in the related organizations for similar applications. 2. Materials and Methods 2. 1. Study area Abbas Abad River is 18 km long from the slopes of Mount Fakhrabad of a height 3312 meters 12 km southwest of Hamedan flowing to the north. The river in its path (between regions Ganjnameh and Abbas Abad), gets branches, and after passing Hamadan Abbas Abad village garden pours in to, in Grachqa Lands into the river (Gishin) khaku. Abbas Abad River, because is a mountainous basin, and the steep, has a swift flow. Due to the snow catchment, this river has a permanent origin. 2. 2. Research Methodology In this study, was used QUAL2KW model to simulate DO, BOD, COD, nitrate, ammonium and phosphate and calculate parameters of self-PURIFICATION capacity of the Abbas Abad River. To calibrate the model, the necessary information for the years 2012-2013 was obtained from the Environmental Protection Agency Hamadan. In order to test and validate the model, data collection was carried out in May, June and August 1394. In order to determine normal flow of the river, two patterns of the three-year river flow continuity and moving average of the river flow. And according to the 43-year period (1971-2013), the normal flow for months of the study was determined. 2. 3. Collecting data According to the field observations and determination of the locations of wastewater disposal into the Abbas Abad River, water harvesting (dividing the water 1 and 2), lateral branches (Tarykdrh) added, were selected in 5 stations sampled on the Abbas Abad River from the Ganjnameh to the Natural History Museum (behind Faculty of agriculture), 7. 43 kilometers long. These stations were sampled and analyzed in the months and required quality tests were conducted. To model, the river was divided into 16 pieces. Since the water quality of the river is influenced by flow and temperature, to determine the critical month in terms of water quality, based on 3-year moving average and Discharge River was 43-year-old took place, who was selected in August as dry months. 2. 4. Self-PURIFICATION capacity and TMDL process In this study, given Abbas Abad River plays apart to supply water to Hamedan city and another part is used as water right of farmers to irrigate gardens and agricultural land, to use drinking water and agriculture standards in the examination of self-PURIFICATION capacity river, two interval were considered. First interval, the range of Ganjnameh up to water division 1, to harvest drinking water and second interval of the division of water up to the water withdrawals for agriculture in behind the School of Agriculture (water division 2) were selected. In this regard, Iran's water quality standards were used. And the standard in order to apply the water used in agriculture FAO standard was used. To determine self-PURIFICATION capacity of river and total maximum of daily load, the TMDL process was used and according to the type of use (agriculture and drinking), in compliance with the respective standards safety margin was considered. For phosphate, the entering load pollutants was reduced up to standard. 3. Results Based on these results, levels of dissolved oxygen in the range of one kilometer beginning to change is not considerable, but with the continue the path up to km 4. 100, relative to the initial state, the average level A drop in 1. 5 percent relatively minor exists, and then up to the end of the path was increased to 5 percent. According to the charts of ammonia and nitrate, with the increasing river discharge and dissolved oxygen, process of nitrification taken place and oxygen caused the conversion of ammonium to nitrate. This increase of nitrate, in the interval (3. 500 km), an average of 15 percent, but is not the index. Also the flow increased that is combined with the relative increase oxygen, enhanced the quality of river water, and dropped amounts of BOD and COD an average of 45 percent. This also causes the status of phosphate to improve in the river. Self-PURIFICATION capacity of river: For drinking purposes, if the values of BOD, COD, NH4 and NO3 increase up to level 1, 0. 3, 9. 3 and 10, fold respectively, they are still acceptable In terms of drinking water standards. It should be noted that this is accordance with position and place of harvest of the river for drinking purposes (the first interval). If in future, the harvesting place of drinking water is changed, the assessment will not be valid. It should be noted that the amount of Phosphate River is always higher than the standards for drinking purposes and thus should be reduced to the initial value of 0. 64. Also, for agricultural purposes, if the values of BOD, COD, NH4, NO3 and PO4 respectively up to 35, 28, 25, 46 and 40-fold increase, still will be accepted in terms of agricultural standards. As already stated, this would suit position and harvesting of water from rivers for agriculture (second period) is valid. If the relative increase parameters, examined compared to relevant standards, we find that the BOD further increases other elements, although significant difference cannot be observed between respective elements, also, there was determined the lowest level of self-PURIFICATION in the first interval of COD and the second interval of NO3. 4. Conclusion and recommendations The aim of this study was to determine assimilative capacity of the Abbas Abad River of Hamedan, in compliance with the standards and quality criteria such as parameters of DO, BOD, COD, NH4, NO3 and PO4. In this regard, QUAL2KW one-dimensional model and the measured data in May, June and August 2011, 2012 and 2015 were used. Was applied the TMDL process for to determine the self-PURIFICATION capacity of the river and total maximum daily load. The results showed that due to the mountainous and steep area, re-aeration is properly done so that although the values of BOD, COD, NH4 and NO3 increase up to 1, 0. 3, 9. 3 and 10-fold respectively, they are acceptable in terms of standards for drinking. The river PO4 is always higher than the standards for drinking purposes and it is necessary to decrease the initial value of 0. 64. For agricultural purposes, if the values of BOD, COD, NH4, NO3 and PO4, respectively, and up to 35, 28, 25, 46 and 40-fold, they will be accepted in terms of agricultural standards. Based on these results, self-PURIFICATION capacity of the river in the first interval (drinking) is more than that in the second interval (agriculture). Also pattern of behavior of DO, BOD, COD, NH4, NO3 and PO4 parameters in river follows a normed probabilistic distributions, normal, Pearson, log normal, Pearson and normal respectively, and this information can help in the prediction of variability in this quality index.

Background: Streptokinase (SK) is an effective and specific thrombolytic treatment of acute myocardial infarction. Despite its significant limitations, streptokinase remains the drug of choice particularly in countries with poorer economies because of its relatively low cost. In this study, the production and PURIFICATION of streptokinase using a pMAL expression vector were evaluated.Methods: The pMAL vector, including the skc gene, obtained from Streptococcus equisimilis H46A, was transformed into E. coli BL21 to produce the soluble active fusion protein SK-MBP. The conditions of SK production were optimized by manipulating temperature, induction time and IPTG concentrations. This protein was purified by DEAE-sepharose column chromatography and the final purity was determined and activity of purified SK-MBP was measured using a synthetic substrate (S2251).Results: After optimizing the production conditions, SK-MBP was the major portion of total protein. Purified SK-MBP formed a single band using SDS-PAGE and had high biological activity.Conclusion: In this study we used pMAL expression vector to produce SK-MBP in E. coli BL21. Using this method we prevented the accumulation of inclusion bodies in spite of the high level of production of SK-MBP. Choosing a suitable host organism for the production of recombinant proteins is one of the most important factors that influence the level of desired protein production. Further studies are recommended to test other host organisms for this purpose.