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Seed aging is a process that can lead to a complete loss of seed viability. This process occurs when seeds are exposed to long-term storage or controlled deterioration treatments such as excessive dryness, lack of oxygen, and temperature fluctuations on Earth or in outer space. Proteomic changes can occur in the dry state of seeds. Extreme temperature fluctuations in outer space, especially on the moon and Mars, are one of the most important challenges for space scientists to transfer plant seeds and grow them in outer space outside the atmosphere. The present study studied the effects of simulated space temperature fluctuations on the quantitative and qualitative content of dry tomato seed proteins. The results showed 13 protein bands in the molecular weight range of 8.89 to 91.82 kilodaltons. A double or more significant decrease in the intensity of protein bands with high molecular weight was also observed in the treated group compared to the control. Finally, the two-dimensional electrophoresis technique followed by mass spectrometry is suggested for better resolution of tomato seed proteins and their better separation, as well as identification of unknown protein bands in future studies.

S-layer proteins of Deinococcus radiodurans are the best self-assemble systems among other proteins that have an essential role in the fabrication of nanowires. Therefore, the purification of these proteins is necessary. The purpose of this research was to optimize the purification of s-layer protein from D. radiodurans with the response surface method. The three factors of SDS concentration, incubation time and mass percent in five levels were considered, and 20 runs were designed by Design-Expert software with a central composite method. Each run includes microbe culture, mass cell preparation, microbe incubation in specific SDS concentration and time and mass percent, separation of the bacteria from detergent with a centrifuge at 5000g, sedimentation of s-layer proteins from detergent solution with a centrifuge at 20000g, determination of protein concentration, and protein purity by Bradford and SDS-PAGE methods, respectively. Finally, the data obtained were analyzed. Analysis of the results demonstrated that at the 95% confidence level, the effect of the detergent concentration factor on the purified protein percent was more than other factors. The optimization results of factors are 5.64% SDS concentration, 7.33% mass percent, and 3 hours incubation time. At optimized conditions the protein concentration and purity percent were obtained 0.584 mg/ml and 47.61% respectively.

Introduction: Triticale (X. Triticosecale Wittmack) was created by doubling the number of F1 hybrid chromosomes between wheat (Triticum spp.) as the female parent and rye (Secale spp.) as the male parent. Investigation of the plants reaction to water shortage is systematically considered by identifying features that are related to drought tolerance subjected to the physiological, cellular, biochemical, and molecular bases. One of the main subjects regarding plants damage under drought stress is the accumulation of reactive oxygen species. Plants possess enzymatic and non-enzymatic systems to prevent oxygen free radical damage which are using them in an adaptive way under drought stress condition. Changes in the expressional level of antioxidant enzymes and the concentration of these enzymes, as well as increasing their activity, are among the very important changes occurring in the cellular level as the result of facing drought stress. Considering the role of the superoxide dismutase enzyme in regulating the amount of oxygen free radicals in plant cells, the aim of the present study was to investigate the gene expression of the superoxide dismutase enzyme with copper/zinc coenzyme under drought stress conditions.Materials and methods: Two genotypes of triticale (sensitive and tolerant) were subjected to drought stress conditions after establishment, and their aerial parts and roots were sampled at 0, 12, 36, and 72 hours after applying drought stress. Drought treatment was applied by weight based on the soil capacity of the pots and by daily weighing of the pots. Each time point repeated three times where the plant shoot was samples prior to plant root. This experiment was carried out in a factorial manner and with a completely randomized design in the greenhouse. The expression level of the superoxide dismutase gene with copper/zinc coenzyme was measured in the samples using the quantitative PCR method.Results: The results of the present study showed that the expression of the Cu/Zn-SOD gene is affected and changed under stress conditions. In general,, with the increase in the amount of stress severity and time of stress, the amount of immediate gene expression increased. The changes in the expression of this gene were greater in the shoots than in the roots, and the shoots responded faster to drought stress. Also, between the two sensitive and tolerant genotypes used in this experiment, the level of Cu/Zn-SOD gene expression in both roots and shoots was higher in the tolerant genotype than in the sensitive triticale genotype.Conclusion: According to the changes in expression and amount in two sensitive and tolerant triticale genotypes, it was generally determined that the superoxide dismutase enzyme is an essential enzyme in the plant to respond to drought stress and also to create tolerance in the triticale plant. Also, according to the changes in gene expression related to the Cu/Zn-SOD isozyme in triticale, it was determined that this isozyme is probably one of the active isozymes of the superoxide dismutase enzyme, whose expression is increased under drought stress conditions.

Introduction: Triticale (X. Triticosecale Wittmack) was created by doubling the number of F1 hybrid chromosomes between wheat (Triticum spp.) as the female parent and rye (Secale spp.) as the male parent. Investigation of the plants reaction to water shortage is systematically considered by identifying features that are related to drought tolerance subjected to the physiological, cellular, biochemical, and molecular bases. One of the main subjects regarding plants damage under drought stress is the accumulation of reactive oxygen species. Plants possess enzymatic and non-enzymatic systems to prevent oxygen free radical damage which are using them in an adaptive way under drought stress condition. Changes in the expressional level of antioxidant enzymes and the concentration of these enzymes, as well as increasing their activity, are among the very important changes occurring in the cellular level as the result of facing drought stress. Considering the role of the superoxide dismutase enzyme in regulating the amount of oxygen free radicals in plant cells, the aim of the present study was to investigate the gene expression of the superoxide dismutase enzyme with copper/zinc coenzyme under drought stress conditions. Materials and methods: Two genotypes of triticale (sensitive and tolerant) were subjected to drought stress conditions after establishment, and their aerial parts and roots were sampled at 0, 12, 36, and 72 hours after applying drought stress. Drought treatment was applied by weight based on the soil capacity of the pots and by daily weighing of the pots. Each time point repeated three times where the plant shoot was samples prior to plant root. This experiment was carried out in a factorial manner and with a completely randomized design in the greenhouse. The expression level of the superoxide dismutase gene with copper/zinc coenzyme was measured in the samples using the quantitative PCR method. Results: The results of the present study showed that the expression of the Cu/Zn-SOD gene is affected and changed under stress conditions. In general,, with the increase in the amount of stress severity and time of stress, the amount of immediate gene expression increased. The changes in the expression of this gene were greater in the shoots than in the roots, and the shoots responded faster to drought stress. Also, between the two sensitive and tolerant genotypes used in this experiment, the level of Cu/Zn-SOD gene expression in both roots and shoots was higher in the tolerant genotype than in the sensitive triticale genotype. Conclusion: According to the changes in expression and amount in two sensitive and tolerant triticale genotypes, it was generally determined that the superoxide dismutase enzyme is an essential enzyme in the plant to respond to drought stress and also to create tolerance in the triticale plant. Also, according to the changes in gene expression related to the Cu/Zn-SOD isozyme in triticale, it was determined that this isozyme is probably one of the active isozymes of the superoxide dismutase enzyme, whose expression is increased under drought stress conditions. [ABSTRACT FROM AUTHOR]

In order to investigate the interaction effects of cadmium as a heavy metal and salicylic acid as an antioxidant and anti-stress substance on sunflower plants, sunflower seeds were washed with distilled water after disinfection with 1% sodium hypochlorite solution and were planted in pots containing sandy soil in a growth chamber with a temperature of 34 ℃ during the day and 25 ℃ at night, and 16/8 hours of light/dark photoperiod. This research was performed as a factorial completely randomized design with four replications under greenhouse conditions. Sunflower plants were exposed to cadmium chloride treatment (0, 50, 100, 150 and 200 µM) at two-leaf stage on a weekly basis. One week after cadmium treatment, plant leaves were sprayed with salicylic acid (0, 250 and 500 µM). After harvesting the plants, cadmium concentrations in roots and shoots were measured. Then, antioxidant enzymes were assayed. Total protein content of leaf samples was extracted and the pattern of leaf protein changes was investigated by SDS-PAGE electrophoresis. Cadmium concentration in roots and shoots increased with increasing cadmium and salicylic acid treatment. Catalase enzyme activity decreased with increasing cadmium concentration, and salicylic acid had a very small effect on its activity. The activity of superoxide dismutase, ascorbate peroxidase, and glutathione reductase enzymes increased with increasing cadmium concentration while increasing salicylic acid concentration reduced the activity of these enzymes. Leaf protein electrophoresis under cadmium and salicylic acid treatments showed differences in leaf protein intensity compared with control plants. The expression of some proteins was increased or decreased in the treated plants. Therefore, cadmium and salicylic acid treatments made changes in the amount of cadmium uptake, antioxidant enzymes activity, and the pattern of sunflower leaf proteins.

The present study was carried out to provide more information about proteins in uterine secretions in the camelus dromedarius. Camel ovaries, blood samples and uterine secretions from 50 sexually mature non-pregnant camels (C. dromedarius) were collected at slaughter. The total protein values of uterine secretions were determined by Lowry method and electrophoresis of proteins on polyacrylamide gel was carried out. Significant fractions of camel uterine proteins were determined. Standard molecular weight comparison revealed that the major protein in uterine fluid had a relative molecular weight of 70 KD and could be albumin. The nearly 30 KD band could be uteroferin and 13-14 KD band corresponded with 14 KD basic polypeptide of a member of plasmin/trypsin inhibitors. On the other hand, the nearly 44 and 23 KD bands were luteal-stage specific. Interestingly the nearly 23 KD luteal phase specific band in this study corresponds with the range of progesterone dependent retinol binding acidic proteins. Although camel uterine proteins are shown to be irrelevant of serum steroids, but the presence of two luteal phase specific bands showed that proteins in uterine secretions may change when the corpus luteum present. [ABSTRACT FROM AUTHOR]

Peanut (Arachis hypogaea) is one of the most important edible oil grains of legume in the world, that understanding the genetic variation among its varieties in choosing the variation and transferring the useful genes is valuable.To study the genetic relationships and diversity of peanut varieties, 10 varieties of peanuts were planted in a research farm in Lasht-Nesha (Guilan) in a randomized complete block design with three replications in 2016. After harvesting the seeds peanut varieties, soluble protein was performed by the Bradford method and SDS-PAGE electrophoresis of seed proteins. In survey of the soluble protein, a local variety of Guilan (Goli) was in group A alone Separately, than the other varieties. Analysis of protein bands with Total lab software showed 17 bands visible on the vertical gel. The highest level of expression of protein was band 11 with a molecular weight of 26 kDa were observed in all genotypes. Cluster analysis of protein bands by Ward method and cut it in 50% similarity, divided genotypes in four groups. Genotypes 6, 7, 8, 9 and 10 in the first group, genotypes 3, 4 and 5 in the second group and genotypes 1 and 2 were separately in groups third and fourth. In general, the results showed that enough diversity in the seed proteins was observed among peanut varieties.

Trichoderma species have been famous for production of cellulases with relatively high enzymatic activity. However, attempts to use their cellulolytic enzymes in the bioconversion of cellulosic wastes have not been successful because of high cost of production and low enzymatic yields. This study aimed to obtain gamma-induced mutants of T. viride with enhanced extracellular cellulase production. Spore suspensions were exposed to γ-rays at 250 Gy as optimum dose. After irradiation, all germinated spores were grown onto PDA plates and mutant strains with better sporulation were selected and subcultured five times to test their stability. Cellulase activity was tested using Whatman No. 1 filter paper, carboxymethyl cellulose, avicel, bacterial cellulose and walseth cellulose according to the IUPAC recommendation. Extracellular proteins profiles of mutant strains were studied via SDS-PAGE. The maximum activity of total cellulase and avicelase were observed in the isolate of M21 (92.43 and 74.40 U/mg, respectively) and maximum endo-glucanase activity was observed in M18 mutant. The results of this study showed that the application of gamma ray led to a significant increase in Cellulose activity of 38 percent of mutant strains. Thus, this method could be used as a simple and efficient way to achieve strains with the ability to produce high levels of enzymes and other biological metabolites.

Background: Prostate cancer is the most common cancer in men. Identification of cancer nature, progress and the mechanism of its inhibition have been always scientific important issues. The aim of this study was to analyze the effect of olives Oleuropein particles on protein expression levels in prostate cancer cell line, LNcap. Methods: Survival test (MTT) has been applied in 8 concentrations of Oleuropein: 19, 39, 78, 156, 312, 625, 1250, and 2500 in LNcap cell line, as well differential comparative proteomics analyses has been performed. Two dimensional electrophoresis results in relation to differential proteins have been identified by MS- MALDI-TOFTOF approach, as their aligns have been recognized in Mascot Database. Results: Survival test demonstrates that olive Oleuropein controled LNCap cells growth and increased their roundness. LNCap cells have most been controlled in 625ppm concentration. GSTP1and DMNT1proteins have been identified as differential proteins in Oleuropein treated samples. Where comparing to controls, DMNT1 had been downregulated. On the other hand GSTP1 had been upregulated in 625 ppm Oleuropein treated samples. Conclusion: Oleuropein may be effective for inhibition of prostate cancer cells in LNcap cell line, by affecting protein expression, antioxidant genes expression, some transcriptional factors expression in relation with cells leading toward apoptosis direction, and metastasis activity reduction. [ABSTRACT FROM AUTHOR]

Background: Finding cures with herbal origins with an easy and cost-effective way of extraction for treatment of diseases such as cancer is highly appreciated. Considering that proteases are involved in initiation and spreading of cancer cells, proteases inhibitors can be take into account as an option for treatment. Sporamin is belonging to trypsine inhibitors group which exists in sweet potato tuber. In this study, the method of sporamin extracting was optimized, and for the first time, its effect on proliferation of breast cancer, MCF-7 cell line, was examined. Methods: Aqueous extraction was obtained from sweet potato. Transparent extraction was inserted into chromatography diethylaminoethanol-Sepharose (DEAE-Sepharose) column. After washing with a linear salt gradient, fractions were separated. To determine protein purity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Sporamin inhibitory activity was measured using Laskowaski method, and eventually 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT method) was used to assess the antiproliferative effect of sporamin. Findings: A single band of 25 kDa sporamin was obtained via SDS-PAGE. Its inhibitory effect was measured 800 units in mg/minute. According to the results of MTT method, the amounts of inhibitory concentration 50 (IC50) for 24 and 48 hours were significantly different (P < 0.01). Conclusion: Using single-column ion exchange and linear salt gradient for sporamin purification was successful; and due to its antiproliferative effect on MCF-7 cells, there is hope that this technique could be used for the purification of drug and treatment of disease in future.

Introduction: Luteolin is an important plant extract. This shows anti-inflammatory, inhibition of cell growth, induction of apoptosis and inhibition of metastasis. Gastric cancer is one of the most common cancers in the world. Also metastasis is widespread in this kind of cancer and late diagnosis is a cause of patient's death. Cofilin, actin and nucleoside-diphosphate kinases (NDPK) are important in cell death and metastasis. In this study the effect of luteolin on MKN-45 gastric carcinoma cells was evaluated. Materials and Methods: At first the cells were cultured and then for 48 hours were cultured with the drug concentration. Then the cells where collected and their protein were extracted and analyzed using 2D electrophoresis and separated on the basis of their molecular mass and PI. After that these samples were used for determine the amino acid sequence by MOLDI-TOF. Results: The growth rate of MKN-45 inhibited under influence of luteolin, this means increasing Apoptosis. Based on the results presented here by identification of protein spots by MOLDI-TOF mass spectrometry the expression of three protein cofilin, actin and NDPK has been decreased by luteolin. Conclusion: It seems that the anti- tumoral effect of luteolin is expressed through three proteins cofilin, actin and NDPK in gastric cancer cells

Introduction: Background: Neff regulatory protein as well as structural proteins such as MPER and V3 of HIV-1 is important targets in vaccine studies. Therefore, in the current study, the Nef-MPER-V3 fusion protein was cloned and expressed in prokaryotic expression system that will be evaluated as a candidate for future vaccine studies. Materials and Methods: The Nef-MPER-V3 sequence was synthesized and inserted into pET-28a(+) vector using Not I/HindIII enzymes. Then, the recombinant construct was expressed in BL21 (DE3) strain of E.coli. For optimization of protein expression, several factors such as time of induction, IPTG concentration and optical density (OD) were evaluated. The recombinant protein was purified on a Ni-NTA agarose column and analyzed by 12% SDS-PAGE. Finally, the fusion protein was confirmed by western blotting. Results: The target gene was successfully cloned into the recombinant vector. The best condition for the expression such as; 2 m IPTG, OD600 = 0.7 and growth 5 hours post-induction were determined and the recombinant protein was purified by affinity chromatography in denaturing condition with urea at a concentration of about 300 μg/ ml. The purified fusion protein migrated as a clear band of around 35 kDa in SDS-PAGE and was detectable using anti-Neff antibody in western blotting. Conclusion: Our results showed that the Nef-MPER-V3 protein was successfully expressed in prokaryotic system and purified protein may provide the antigen for future experiments for immunogenicity evaluation in mice is currently undertaken. [ABSTRACT FROM AUTHOR]

Background: Nowadays, more than 100 types of cancer are known worldwide, which leukemia is one of the most common type. Chronic myeloid leukemia (CML) is one of the most recognized forms of leukemia, accounting for between 15% and 20% of total leukemia. Efforts to find new synthetic compounds are one of the basic strategies for finding new drugs for CML treatment. The purpose of this study was to investigate the effects of growth suppression and induction of apoptosis (family of pyrazinoic acid) on the cancer cell line of K562 leukemia. Methods: In this study, the effect of a certain concentration of 2-oxo-1, 2-diphenylethyl-pirazine-2-carboxylate (2 DP.P) was evaluated on the growth, cell cycle and induction of apoptosis of K562 cells using various techniques. MTT test was used to investigate the effects of 2-DP.P on cell viability. For this purpose, 5×104 of K562 cells were cultured at concentrations of 25-100 μM from the 2-DP.P for 24, 48 and 72 hours intervals. Electrophoresis, fluorescence microscopy and flow cytometry were used for analysis of apoptosis. Results: The 2-DP.P inhibited dose and time dependently growth in K562 cells. IC50 of the compound was calculated 25 μM. Also, data from fluorescence microscopy, DNA fragmentation and cell cycle analysis confirmed the reduction of viability and induction of apoptosis by the compound. Conclusion: due to the effects of growth inhibition and induction of apoptosis by the 2-DP.P, this compound can be suggested as a new and effective compound for further studies in treating patients with leukemia. [ABSTRACT FROM AUTHOR]

Background and Aims: Many bacteria including Escherichia coli may enter into a viable but non-culturable (VBNC) state under unfavorable stresses, which are unable to be detected by culture-based methods. In this study, the use of Reverse Transcription PCR (RT-PCR) for detection of VBNC state of E. coli O157:H7 was investigated. Materials and Methods: Escherichia coli O157:H7 was inoculated in distilled water and 30 percent salt water at room temperature. The cultivability of bacteria was determined using routine culture and colony counting on MacConkey agar. The RT-PCR of 16S rRNA gene involved direct extraction and purification of RNA, DNase I treatment for removing DNA contamination, cDNA synthesis and electrophoresis of PCR products of cDNA was used to detect viable E. coli O157:H7 under studied treatments and was compared with the results of RT-PCR of 16S rRNA gene of heat- killed bacteria. Results: The cultivability of bacteria was maintained during the study period in distilled water; however, the use of 30% NaCl caused the bacteria to be non-cultivable on day 4. The RTPCR of 16S rRNA showed the positive expression of this gene in cultivable and non-cultivable bacteria during the study period, whereas heat-killed bacteria were negative for this gene, which indicated the efficacy of RT-PCR of 16S rRNA in differentiation of alive from dead bacteria. Conclusions: Escherichia coli O157:H7 entered into the VBNC under 30 percent NaCl which can be associated with serious human health problems. RT-PCR can be used to detect bacteria in the VBNC state. [ABSTRACT FROM AUTHOR]

Background: Many immunologic and inflammatory mechanisms play a role in asthma etiology. A member of a T cell immunoglobulin (Ig) domain and mucin domain (TIM) gene family, TIM-1, in 5q31-33 region is shown to be correlated with development of T helper-2 (TH2) Cells and allergic diseases. So the aim of this study was to investigate the association of the genotype and allele frequencies of the TIM-1polymorphisms with asthma and the relationships among this polymorphism to IgE levels in patients with asthma. Methods: PCR-RFLP (Polymerase chain reaction–restriction fragment length polymorphism) and polyacrylamide gel electrophoresis (PAGE) were used for investigating the presence of TIM-1 polymorphisms in 200 asthmatic patients and 200 healthy controls. To assess the frequency of genotypes and its relationship with serum immunoglobulin E and asthma, chi-square test was used and P<0/05 was considered significant. Results: The genotype and allele frequency of TIM-1 polymorphisms were significantly different between the patient and controls. Significant association was observed between this polymorphism and the risk of asthma. Conclusion: The 5529A>G, 416G>C and 5529A>G polymorphismS of TIM1 gene may be associated the susceptibility of asthma and also with total serum IgE levels. [ABSTRACT FROM AUTHOR]

Hemoglobinopathies are most common inherited disorders in the world; approximately 7 percent of the worldwide population and 5-6 percent of population of Iran are carriers. The hemoglobin disorders inherit as autosomal recessive and are very common in the Mediterranean area and much of the Asia and Africa. The control of this inherited disorders need to genetic counseling and accurate screening by more advanced and more accurate methods. This study explains features of current Iran hemoglobin disorders, nominates the accessible methods for screening them and introduces the capillary zone electrophoresis as a rapid and more accurate method. The required data were extracted of various articles and then for good explanation, current Iran hemoglobinopathies properties were showed in the tables and electropherograms of important hemoglobin disorders in Iran population were provided for help to interpretation results of blood tests by capillary zone electrophoresis method. Hemoglobin disorders are including thalassemias and hemoglobin variants; Disruption in the production and malfunction of globin chains cause types of hemoglobin disorders. We cannot introduce one of clinical laboratory tests as critical and basic method for screening and distinguishing types of inherited hemoglobin disorders as alone. For distinguishing the types of them must be prepared enough information and data of the hemoglobin disorders and for more accurate analysis must be used simultaneously different methods as gel electrophoresis, high performance liquid chromatography, isoelectric focusing, capillary zone electrophoresis or molecular tests. The capillary electrophoresis is an accurate and rapid method for screening types of the hemoglobin disorders. Other side this method cannot analyze all of them, so must be used biochemical, biophysical and molecular methods for confirmation the results. This review showed we can use the capillary electrophoresis and HPLC as two complementary methods for hemoglobinopathies screening. We can analyze by the methods more hemoglobin disorders and decrease more laboratory errors. Moreover, we must have patient history, hematological indices, information and data of types of hemoglobinopathies. The patient history and complete blood count results as red blood cell count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration can be useful and helpful in screening the hemoglobin disorders and then distinguishing all of hemoglobin disorders.

The genus Salvia (Lamiaceae) comprises over 900 species in the world, with a relatively wide dispersion in the Iran’s flora. Until now, about 58 species of the genus have been reported and identified in Iran, in which 17 of them were endemic. In order to study, investigate and evaluate the intraspecific diversity, similarity and dissimilarity among Iranian Salvia officinalis accessions, an experiment was carried out using SDS-PAGE technique. In this study, the seeds from five accessions were collected from gene bank and were evaluated. The seed storage proteins were extracted by buffers and were measured. Phylogenetic relationships were analyzed according to presence and absence of bands on the gel. A dendrogram was prepared using calculation of the accession’s similarity index. Mean comparison were done by Tukey’s test. The seed protein contents showed significant differences (p≤0.01) among accessions. A total of 39 bands were indicated on the gel. The maximum diversity was detected in the accession No. 2 while, the lowest band’s number were recorded with the accessions No. 1 and 5. Based on dendrogram, the accessions were divided into two groups; one includes accession No. 1 and 4 other accessions were located in the second group further classified into two subgroup including accessions No. 2 and 3 in one clade and accessions No. 4 and 5 in the other ones.

The country Iran is one of the most important centers for diversity of the genus Verbascum. This study was carried out to determinate intraspecific diversity of the V. speciosum populations collected from East Azerbaijan, West Azerbaijan, Kordestan, Kermanshah and Hamedan provinces. With using the D.S.S. method seventeen special habitats were determined for V. speciosum and floristic-ecologic data collected from each special habitat. Seed storage proteins were subjected for electrophoresis studies. In the survey of all special stations, 109 plant species were distinguished as associated species. As the result of analysis of floristic data, as floristic marker, four groups were determined that this grouping is distinguished the existence of intraspecific diversity in V. speciosum. Analysis of ecologic data was also confirmed the above mentioned grouping. The analysis of seed storage protein banding pattern showed four quite obvious groups. In the survey of electrophoresis pattern, existence of differences regarding number and density of the protein bands indicating existence of intraspecific diversity in the populations of V. speciosum. Therefore in this species, the grouping that introduced with floristic marker, mostly confirmed also with ecological and electrophoresis markers. This means that floristic grouping can only be used as a cost-effective and efficient method for studying of intraspecific diversity.

Background: DNA-based computing is an emerging research aspect that enables the in-vivo computation and decision making with significant correctness. Recent papers show that the expression level of miRNAs are related to the progress status of some diseases such as cancers and DNA computing is introduced as a low cost and concise technique for detection of these biomarkers. In this paper, DNAbased logic gates are implemented in the laboratory to detect the level of miR-21 as the biomarker of cancer. Materials and Methods: At the first, required strands for designing DNA gates are synthesized. Then, double stranded gate is generated in laboratory using a temperature gradient that followed by electrophoresis process. This double strand is the computation engine for detecting the miR-21 biomarker. miR-21 is as input in designed gate. At the end, the expression level of miR-21 is identified by measuring the generated fluorescent. Results: at the first stage, the proposed DNA-based logic gate is evaluated by using the synthesized input strands and then it is experimented on a tumor tissue. Experimental results on synthesized strands show that its detection quality/correctness is 2.5x better than conventional methods. Conclusion: Experimental results on the tumor tissues are successful and are matched with those are extracted from real time PCR results. Also, the results show that this method is significantly more suitable than real time PCR in view of time and cost. [ABSTRACT FROM AUTHOR]

Application of lysozyme as a natural antimicrobial or preservative agent in food and pharmaceutical industry is well known. The antimicrobial effects of lysozyme can be expanded by conjugation with carbohydrates via Maillard reaction, however this natural reaction is a time consuming process. So in this study a new method microwave heating, was applied to Accelerate the Maillard reaction. Lysozyme was mixed with dextran and heated by microwave irradiation. The glycation extent was determined by SDS-PAGE electrophoresis, cation-exchange chromatography and sugar analysis. SDS-page pattern show that with increasing heating time a high molecular weight conjugates were appeared and maximum glycosylation was occurred after 4 minutes heating. The elution profiles (chromatograms) of the lysozyme–dextran conjugates compared the natural lysozyme showed that the conjugated peak shifted towards the higher molecular weight fraction and overlapped with the non conjugated proteins, indicated that lysozyme was covalently bound to dextran. These results suggest that Maillard reaction rate can be increase using microwave heating. [ABSTRACT FROM AUTHOR]

Background and Aims: Brucellosis is an infectious zoonotic disease responsible for reproductive disorders among animals and significant economic losses to the meat and milk supplychain. The purpose of this study was therefore to evaluate and diagnose the species and strains of Brucella isolates among cattles via multiplex polymerase chain reaction. Materials and Methods: In this study, blood samples were taken from 22 cows diagnosed as having brucellosis by PCR test. Blood samples were cultured in BACTEC vials and incubated at 37 degrees Celsius for 5 days following which each sample was cultured on Brucella agar and kept for 3 days under Co2 condition.The 16SrRNA gene was used as the target for the detection of Brucella genus and the bcsp31 gene was used for the detection of Brucella biovars. The PCR products were electrophoresed on %1 agarose and compared with standard strains. Results: The PCR results showed a total of 22 positive samples for Brucella among cattles. Out of the 22 cattle positive samples, 19 were identified as B.abortus (biovars 1, 2, 3, 4, 5) and 3 as B. melitensis (biovars 1, 2, 3). Conclusions: Brucella abortus biovars 1 and 3 and B .melitensis biovar 1 are the most prevalent biovars among cattles in Lorestan province, Iran. Moreover, multiplex PCR method could be used to improve the qualitative level and the Brucella diagnosis speed in laboratories. [ABSTRACT FROM AUTHOR]

Background and Objectives CD40L is a membrane protein and a member of the tumor necrosis factor super family (TNFSF) that plays an important role in transferring cell signaling in innate and adaptive immunity. Since this protein plays its role through clustering the receptors on the target cell surface, it seems that the multimeric form of this molecule can have a stronger effect than the natural trimeric form. So in this study we describe cloning and expression of dodecameric soluble CD40L through self multimerizing protein, surfactant protein D (SP-D), in the HEK293 cell line. Materials and Methods In this experimental study, the SPD-CD40L was designed in silico and cloned in pcDNA3.1(+) plasmid. HEK293 cells were transfected and used as the expression host cells. The expression of SPD-CD40L was determined at the RNA level by RT-PCR method. After the purification of the recombinant proteins by affinity chromatography, the molecular weight of the recombinant protein was determined by SDS-PAGE. To evaluate the specificity of protein, ELISA and Dot Blot were used. Results The results indicated that HEK293 cells were transfected and recombinant protein was expressed 24 and 48 hours after Transfection. Moreover, the results of ELISA and Dot Blot methods represented the specificity of recombinant SPD-CD40L chimeric protein. Conclusions Chimeric protein SPD-CD40L was expressed successfully by pcDNA3.1(+) in eukaryotic HEK293 cell line and the molecular weight of 4-trimeric (dodecameric) protein was shown to be consistent with the expected amount. [ABSTRACT FROM AUTHOR]

Background and Objectives There is considerable phenotypic variation of Beta-thalassemia with common mutations whose understanding would facilitate the prevention of the specific syndromes. The aim of this study was to determine the common Beta-thalassemia mutations in Isfahan and Chaharmahal Bakhtiari province and thier relationship with blood parameters. Materials and Methods In this descriptive study, 10 ml venous blood samples were taken from 321 Beta thalassemia carriers having referred to the private Isfahan Center of Medical Genetics in 2016. For the confirmation of the diagnosis of Beta thalassemia in these patients, the indices of MCV and MCH with Mindary device and HbAl, HbA2, Hbf and RBC with the electrophoresis method were measured, yf, t-test and SPSS 22 were used in analyzing data. Results Mutation Fr36/37 (-T) in the population studied in Isfahan and Chaharmahal Bakhtiari with the frequency rates of 34 (26.35%) and 22 (32.35%) showed the highest in the studied mutations. About 80% of cases were detectable mutations in the beta globin gene in the people with HbA2 > 3.5 and in 100% of cases with MCH < 27 and MCV < 80. Conclusions Beta-thalassemia mutations among Chaharmahal Bakhtiari and Isfahan populations show diversity and wide distribution. Average mutations studied based on blood indices showed a wide variety. The results showed a positive correlation between beta-thalassemia mutations and red blood cells indices which can be effective in fast and efficient screening of these common mutations. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a protozoan parasitic disease and a global health problem. The aim of this study is to diagnose the parasitic infection in humans for epidemiological identification and providing control programs using proprietary co-designed primers in three species of Leishmania. Materials and Methods: 30 common Leishmania isolates were gathered from different centers in Iran. Having been cultured in RPMI-1640 Medium, DNA was extracted and the gene ITS2-rRNA was amplified by PCR. The amplicons were examined by electrophoresis on agarose gel 2%. Also, in FLASH PCR method, a specific probe and florence colour were used to investigae the amplicon existence on sample. Results: The results of the investigations by PCR and FLASH PCR methods show that these methods are sensitive and specific for diagnosis of Leishmania Conclusion: In this study, identification of Leishmania parasite using specific primer pairs was successful and TaqMan could be one of the most sensitive diagnostic methods to identify parasite load for the ITS2 region of Leishmania [ABSTRACT FROM AUTHOR]

Introduction: Nowadays, rising of cancer prevalence has increased the importance of its treatment. Leukemia is the cancer of blood-forming tissues which derives from red and white blood progenitors cells. Prednisolone (Family members of glucocorticoid drugs) with anti-tumor and apoptotic effects has been considered by many researchers. Objective: The aim of this study was the investigation of the effect of prednisolone on K562 cell line. Materials and Methods: First, The K562 cells were cultured. After, by using different concentrations of prednisolone, antitumor properties of this drug on K562 cells was measured by MTT assay after 72 hours. Finally, Hoechst (33342) staining and DNA electrophoresis were used for study of apoptosis. Data analysis was performed using SPSS software and ANOVA test (P<0.05). Results: In this study, anti-tumor effect of prednisolone on K562 leukemia cell line was confirmed. The results also showed that prednisolone, compared with the control group, had cytotoxic activity that is highest in 100μM at 72 hours.Statistical analysis indicated that this drug inhibited the growth of K562 cells (P < 0.05). Conclusion: Based on the results, prednisolone is an effective agent for chronic myeloid leukemia therapy and prevention. We suggest that further studies on the effects of this drug on animal models and human be done. [ABSTRACT FROM AUTHOR]

The genus Salvia (Lamiaceae) comprises over 900 species in the world, with a relatively wide dispersion in the Iran's flora. Until now, about 58 species of the genus have been reported and identified in Iran, in which 17 of them were endemic. In order to study, investigate and evaluate the intraspecific diversity, similarity and dissimilarity among Iranian Salvia officinalis accessions, an experiment was carried out using SDS-PAGE technique. In this study, the seeds from five accessions were collected from gene bank and were evaluated. The seed storage proteins were extracted by buffers and were measured. Phylogenetic relationships were analyzed according to presence and absence of bands on the gel. A dendrogram was prepared using calculation of the accession's similarity index. Mean comparison were done by Tukey's test. The seed protein contents showed significant differences (p≤0.01) among accessions. A total of 39 bands were indicated on the gel. The maximum diversity was detected in the accession No. 2 while, the lowest band's number were recorded with the accessions No. 1 and 5. Based on dendrogram, the accessions were divided into two groups; one includes accession No. 1 and 4 other accessions were located in the second group further classified into two subgroup including accessions No. 2 and 3 in one clade and accessions No. 4 and 5 in the other ones. [ABSTRACT FROM AUTHOR]

Hemoglobinopathies are most common inherited disorders in the world; approximately 7 percent of the worldwide population and 5-6 percent of population of Iran are carriers. The hemoglobin disorders inherit as autosomal recessive and are very common in the Mediterranean area and much of the Asia and Africa. The control of this inherited disorders need to genetic counseling and accurate screening by more advanced and more accurate methods. This study explains features of current Iran hemoglobin disorders, nominates the accessible methods for screening them and introduces the capillary zone electrophoresis as a rapid and more accurate method. The required data were extracted of various articles and then for good explanation, current Iran hemoglobinopathies properties were showed in the tables and electropherograms of important hemoglobin disorders in Iran population were provided for help to interpretation results of blood tests by capillary zone electrophoresis method. Hemoglobin disorders are including thalassemias and hemoglobin variants; Disruption in the production and malfunction of globin chains cause types of hemoglobin disorders. We cannot introduce one of clinical laboratory tests as critical and basic method for screening and distinguishing types of inherited hemoglobin disorders as alone. For distinguishing the types of them must be prepared enough information and data of the hemoglobin disorders and for more accurate analysis must be used simultaneously different methods as gel electrophoresis, high performance liquid chromatography, isoelectric focusing, capillary zone electrophoresis or molecular tests. The capillary electrophoresis is an accurate and rapid method for screening types of the hemoglobin disorders. Other side this method cannot analyze all of them, so must be used biochemical, biophysical and molecular methods for confirmation the results. This review showed we can use the capillary electrophoresis and HPLC as two complementary methods for hemoglobinopathies screening. We can analyze by the methods more hemoglobin disorders and decrease more laboratory errors. Moreover, we must have patient history, hematological indices, information and data of types of hemoglobinopathies. The patient history and complete blood count results as red blood cell count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration can be useful and helpful in screening the hemoglobin disorders and then distinguishing all of hemoglobin disorders. [ABSTRACT FROM AUTHOR]

Background and purpose: The purpose of this study was to design a recombinant vector pEFGP- N1 containing the full length of HIV-1Vpr gene. To the best of our knowledge, the cloning of Vpr gene in pEGFP_N1 is not previously done. Materials and methods: As a source of Vpr gene the pUC19-Vpr recombinant vector was confirmed by digestion with restriction enzymes BglII and NotI in order to separate the Vpr gene. The specific primers for Vpr gene including the restriction sites of XhoI and KpnI were designed according to the multiple cloning site of pEGFP-N1 and PCR reaction was performed using the pUC 19-Vpr vector as a template. The PCR product was undergone electrophoresis and gel extraction. Digestion reaction was done on both the extracted PCR product and the pEGFP-N1vector. The recombinant pEGFP-N1-Vpr vector was achieved by the ligation reaction using the T4 DNA ligase and it was transformed into the E-coli (DH5α) and propagated. Finally, confirmation was done through the restriction enzyme digestion and PCR amplification. Results: The recombinant vector pUC19-Vpr was confirmed using the restriction enzymes digestion, and then Vpr gene was successfully amplified using its specific primers including restriction sites for XhoI and KpnI. The PCR product was confirmed by electrophoresis. Finally, a recombinant vector pEGFP-N1 containing the full length of human immunodeficiency virus-1 Vpr gene (pEGFP-N1-Vpr) was successfully constructed. Conclusion: HIV-1 Vpr gene in its full length size could be ligated into the pEGFP-N1. [ABSTRACT FROM AUTHOR]

Background and Objective: Acute myeloid leukemia (AML) is a clonal proliferation of immature myeloid progenitors in the bone marrow. Multiple genetic and environmental factors have been reported to be involved in the pathogenesis of AML. Cytochromes, such as cytochrome P450 are among detoxifying enzymes whose aberrant expression can make individuals susceptible to a variety of cancers and leukemias including AML. The aim of this study was to evaluate the G15631T, CYP2B6 subfamily of cytochrome P450 gene with susceptibility to AML in some AML patients and normal individuals. Materials and Method: We enrolled 76 AML patients and 80 normal individuals as the control group for the study of G15631 T polymorphism of CYP2B6 gene. This polymorphism was amplified by PCR using specific primers. The PCR products were digested by Bsr I enzyme and Electrophoresed on Agarose gel. Results: The frequency of G15631T polymorphism was significantly higher in patients (55%) than the control group (31%). The risk of AML (with CI = 95%) in patients with GT genotype was 1.95 fold more than individuals with the same genotype in the control group. The TT genotype was found in 4 patients with AML, but in none of the individuals in control group. Conclusion: The results show that there is a direct relationship between G15631T, CYP2B6 gene polymorphism and susceptibility to acute myeloid leukemia in some patients in our study population. However, further studies on larger populations are recommended. [ABSTRACT FROM AUTHOR]

The country Iran is one of the most important centers for diversity of the genus Verbascum. This study was carried out to determinate intraspecific diversity of the V. speciosum populations collected from East Azerbaijan, West Azerbaijan, Kordestan, Kermanshah and Hamedan provinces. With using the D.S.S. method seventeen special habitats were determined for V. speciosum and floristic-ecologic data collected from each special habitat. Seed storage proteins were subjected for electrophoresis studies. In the survey of all special stations, 109 plant species were distinguished as associated species. As the result of analysis of floristic data, as floristic marker, four groups were determined that this grouping is distinguished the existence of intraspecific diversity in V. speciosum. Analysis of ecologic data was also confirmed the above mentioned grouping. The analysis of seed storage protein banding pattern showed four quite obvious groups. In the survey of electrophoresis pattern, existence of differences regarding number and density of the protein bands indicating existence of intraspecific diversity in the populations of V. speciosum. Therefore in this species, the grouping that introduced with floristic marker, mostly confirmed also with ecological and electrophoresis markers. This means that floristic grouping can only be used as a cost-effective and efficient method for studying of intraspecific diversity. [ABSTRACT FROM AUTHOR]