Sensitive high-resolution melting analysis for screening of kras and braf mutations in Iranian human metastatic colorectal cancers

Background: Investigations of methods for detection of mutations have uncovered major weaknesses of direct sequencing and pyrosequencing, with their high costs and low sensitivity in screening for both known and unknown mutations. High resolution melting (HRM) analysis is an alternative tool for the rapid detection of mutations. Here we describe the accuracy of HRM in screening for KRAS and BRAF mutations in metastatic colorectal cancer (mCRCs) samples. Materials and Methods: A total of 1000 mCRC patients in Mehr Hospital, Tehran, Iran, from Feb 2008 to May 2012 were examined for KRAS mutations and 242 of them were selected for further assessment of BRAF mutations by HRM analysis. In order to calculate the sensitivity and specificity, HRM results were checked by pyrosequencing as the golden standard and Dxs Therascreen as a further method. Results: In the total of 1,000 participants, there were 664 (66.4) with wild type and 336 (33.6) with mutant codons 12 and/or 13 of the KRAS gene. Among 242 samples randomly checked for the BRAF gene, all were wild type by HRM. Pyrosequencing and Dxs Therascreen results were in line with those of the HRM. In this regard, the sensitivity and specificity of HRM were evaluated as 100. Conclusion: The findings suggest that the HRM, in comparison with DNA sequencing, is a more appropriate method for precise scanning of KRAS and BRAF mutations. It is also possible to state that HRM may be an attractive technique for the detection of known or unknown somatic mutations in other genes

Mutation analysis of KRAS and BRAF genes in metastatic colorectal cancer: A first large scale study from Iran

Background: The investigation of mutation patterns in oncogenes potentially can make available a reliable mechanism for management and treatment decisions for patients with colorectal cancer (CRC). This study concerns the rate of KRAS and BRAF genes mutations in Iranian metastatic colorectal cancer (mCRC) patients, as well as associations of genotypes with clinicopathological features. Materials and Methods: A total of 1,000 mCRC specimens collected from 2008 to 2012 that referred to the Mehr Hospital and Partolab center, Tehran, Iran enrolled in this cross sectional study. Using HRM, Dxs Therascreen and Pyrosequencing methods, we analyzed the mutational status of KRAS and BRAF genes in these. Results: KRAS mutations were present in 33.6 cases (n=336). Of KRAS mutation positive cases, 85.1 were in codon 12 and 14.9 were in codon 13. The most frequent mutation at KRAS codon 12 was Gly12Asp; BRAF mutations were not found in any mCRC patients (n=242). In addition, we observed a strong correlation of KRAS mutations with some clinicopathological characteristics. Conclusions: KRAS mutations are frequent in mCRCs while presence of BRAF mutations in these patients is rare. Moreover, associations of KRAS genotypes with non-mucinous adenocarcinoma and depth of invasion (pT3) were remarkable

LiNi0.5Mn1.5O4 Thin Films Grown by Magnetron Sputtering under Inert Gas Flow Mixtures as High-Voltage Cathode Materials for Lithium-Ion Batteries

Delivering a commercial high-voltage spinel LiNi0.5Mn1.5O4 (LNMO) cathode electrode for Li-ion batteries would result in a significant step forward in terms of energy density. However, the structural ordering of the spinel and particle size have considerable effects on the cathode material's cyclability and rate capability, which are crucial challenges to address. Here, a novel mid-frequency alternating current dual magnetron sputtering method was presented, using different Ar-N-2 gas mixtures ratios for the process gas to prepare various LNMO thin films with highly controlled morphology and particle size; as determined from X-ray diffraction, Raman spectroscopy and electron microscopy. It resulted in enhanced cycling and rate performance. This processing method delivered N-containing LNMO thin film electrodes with up to 15 % increased discharge capacity at 1 C (120 mAh g(-1)) with respect to standard LNMO (grown under only Ar gas flow) thin film electrodes, along with outstanding rate performance up to 10 C (99 mAh g(-1)) in the operating voltage window 3.5-4.85 V vs. Li+/Li. Besides, electrochemical impedance spectroscopy results showed that the intricate phase transitions present in standard LNMO electrodes were almost suppressed in N-containing LNMO thin films grown under different Ar-N-2 gas flow mixtures

An investigation on possibility of nile tilapia (Oreochromis niloticus) feminization under the condition of brackish water in Bafgh

This study was aimed to investigate the effect of different doses of oral diethylstilbestrol (DES) on feminization of Nile tilapia (Oreochromis niloticus) larvae under the condition of brackish water in Bafgh fisheries research center. This Experiment was comprised of two control groups in two replicates and two experimental ones in three replicates. At the beginning of the trial, a number of 2150 larvae were randomly distributed among 10 replicates that were under treatment for 45 days. The diets in two control groups, i.e. non-treated, C1, and ethanol treated feed, C2, and the ones in two experimental groups, i.e. 200 and 400 mg DES/k of feed, E1 and E2, were administrated to 7 day post fertilization (dpf) larvae for a period of 35 days and then until day 45, the larvae were fed with ordinary feed. At the end of the trial, the gender ratio of larvae in different groups was determined by squash test in 50 randomly selected individuals from each replicate. The results showed no significant differences for gender ratios within control groups (respective male to female ratio in C1 and C2 was 47:53 and 57:43). However, the corresponding ratios within experimental ones, fed with 200 and 400 mg DES/k of feed respectively, was significantly different (P<0.05). Noteworthy that the percent of females between two experimental groups (E1 and E2) was not significantly different (70.0 and 64.7 respectively) but it was significantly different with ratios in experimental groups (P<0.05). The total body length and live body weight were determined at the begging and then at day 30 and 45, it was randomly measured in 20 larvae from each replicate. At the beginning of the experiment, the mean body weight and total length of larvae was 0.012 gram and 1.03 centimeter respectively. The respective averages at day 30 and 45 were significantly different among groups (P<0.05). Survival rate was significantly different among groups (P<0.05), being highest in E1 and E2 respectively whereas lowest in control ones (56.51 and 58.84 in C1 and C2 respectively)

Genetic characterization of Vimba vimba persa in southern part of the Caspian Sea using microsatellite marker

Population genetic structure of Vimba vimba persa was investigated using microsatellite markers from 4 regions along the Iranian coastline of Southern the Caspian Sea (Anzali lagoon & havigh River in Gilan province, BabolRoud River in Maz&eran province & GorganRoud River in Golestan province). Genomic DNA from 121 specimens was extracted from fin tissue by phenol-Chlorophorm method & PCR reaction was accomplished with 17 microsatellite primers, out of 17 microsatellite primers 14 loci were amplified, in which 10 of them were amplified with reasonable polymorphism & 4 were monomorphism. Totally 302 alleles were identified on average 7.5 Observed & expected heterozygosity averages were 0.80 & 0.77 respectively. Most cases significantly deviated from Hardy-Weinberg equilibrium (p 0.01). The estimation of Fst (p 0.01) revealed significant population structuring & estimate four population of Vimba vimba persa is identified in the Caspian Sea. These studies were to apply & develop population genetic approaches to assist conservation, sustainable harvest & restocking of these populations

Identification and extraction of streptococcusis gene from streptococcus iniae and recording in Gene Bank

S. iniae is an important pathogen in both marin animals and humans, causing systemic infections in these hosts. The symptoms of infections are very similar to symptoms of infections caused by human¬specific streptococcal pathogens. The enzyme phosphoglucomutase (PGM) has recently been discovered to play an important role in polysaccharide capsule production and virulence in s. iniae. We aim to isolate S. iniae and cloning phosphoglucomutase gene. S. iniae grew on sheep blood agar as mucoid and beta- hemolytic colonies after 24 h of incubation at 37C. Then confirmed it by sensitive and rapid method of PCR. The pgm gene was amplified successfully and cloned in pTZ57R cloning vector.The recombinant plasmid was sub cloned into pETDuet-l expression vector by rectriction enzymes and confirmed by PCR. Although S. iniae is a pathogen of economic importance, relatively little is known regarding mechanisms of its pathogenesis and few specific virulence determinants have been established. These include the enzyme phosphoglucomutase which contributes to cell wall integrity and resistance to antimicrobial peptides. Thus, this enzyme is the best choice to product the vaccine and more investigations

Population genetic study on croaker species Atrobucca nibe in Persian Gulf and Oman Sea using molecular methods

Black Mouth Croaker (Atrobucca nibe) is of significant value from the economics point of view. In the following study, the genetic diversity of Atrobucca nibe has been investigated using the PCR-sequencing method from three different regions of the Persian gulf and Oman Sea, namely Chabahar in the Sistan and Baluchestan and Jask in Hormozgan Province of Iran and Fujairah in the Fujairah Emirate of the United Arab of Emirates. The purpose of the following exploration has essentially beencoined to further the study of Atrobucca nibe genetic diversity and its population structure in the Persian Gulf and Oman Sea. For this purpose, a number of 54 fish have been collected from the aforementioned regions. The DNA of the fish had been extracted through the Ammonium Acetate method and the quality of the DNAs were analyzed via the spectrophotometry and furthermore the quantity of the extracted DNA were assessed viaelectrophoresis. Polymerase Chain Reaction (PCR) was conducted on the targeted DNAs and thenceforth DNA sequencing was carried out. Subsequently, through sequencing it was discovered that the D-loop region in the mitochondrial DNA (mtDNA) of Black Mouth Croaker,contained 270 base pairs (bp). For the purpose of analyzing the genetic diversity of Atrobucca nibe and its population structure in the Persian Gulf and Oman Sea, the following softwares have been conducted, namely theBioEdit, Arlequin, Dnasp and Mega5 software. Furthermore, the Kimura 2-parameter was used for the purpose of genetic distance analysis. Through observations of average levels of FST between the regions, it can be summed up that genetic differences do exist among the present populations. On the grounds of the obtained results from the Persian Gulf and Oman Sea, it can be concluded that there currently are three different populations of Atrobucca nibe residing in the waters of Sistan & Baluchestan, Hormozgan and Fujairah

Analogue peptides for the immunotherapy of human acute myeloid leukemia

Accepted manuscript. The final publication is available at: http://link.springer.com/article/10.1007%2Fs00262-015-1762-9The use of peptide vaccines, enhanced by adjuvants, has shown some efficacy in clinical trials. However, responses are often short-lived and rarely induce notable memory responses. The reason is that self-antigens have already been presented to the immune system as the tumor develops, leading to tolerance or some degree of host tumor cell destruction. To try to break tolerance against self-antigens, one of the methods employed has been to modify peptides at the anchor residues to enhance their ability to bind major histocompatibility complex molecules, extending their exposure to the T-cell receptor. These modified or analogue peptides have been investigated as stimulators of the immune system in patients with different cancers with variable but sometimes notable success. In this review we describe the background and recent developments in the use of analogue peptides for the immunotherapy of acute myeloid leukemia describing knowledge useful for the application of analogue peptide treatments for other malignancies

Growth performance and age composition of Salmo trutta caspius in Iranian part of Caspian Sea

The aim of this project is to study the growth and age structure in the Caspian trout, comparison with other coldwater species and check the trend of these parameters in recent years. This study was conducted during 2013 till 2015. Totally, 43 specimens for back calculation and 101 specimens for biometrical study of the Caspian trout have been caught in two month period. Biometric parameters such as length, weight and age of the fish were recorded. Base on Back calculation method in 1393, the average length of fish at ages 1, 2 and 3 years old were 18.98 ± 3.5, 30.5 ± 7.24 and 41.7 ± 9.1 cm. So these age groups are under the adult age and don’t approaching to near the beach and rivers for spawning behavior. Therefore, these length groups cannot be observed in catch composition. The result showed, the mean of gonad weight in this fish was about 11 percent of total weight and number of eggs per gram of gonad calculated about 10.8 numbers. Minimum age and maximum age of this species determined 4 years and 7 years (mean = 5.6) and the most frequency allocated to 5 age group and the frequency of 6 and 7 years has been remarkable. The average length of salmon was 69.2 ± 6.2 cm (minimum 57 and maximum 81 cm) and the average weight was measured 3323 ± 677 g (2400 to 5600 g) in the catch composition. Growth parameters such as k, L_∞ and ø’ was measured 0.18, 104 cm and 3.289 respectively. The amount of b for relation length and weight was 2.9 which imply negative allometry. L_∞ and growth coefficient (K) on the Caspian trout were acceptable range, that it shows good growth the fish in the sea water. Most of the fishes were catched from Cheshmehkileh River. As at present Shilat uses just the broods of the Tonekaboon region for restocking of this species, we recommend using the broods of the western region separately for rehabilitation of the stocks of this region

The comprehensive study of possibility ecological control Mnemiopsis leidyi in Caspian Sea activity: The study and recognization parasitic fauna and Bacterial flore in ctenophore (Mnemiopsis leidyi and Bereo ovata)

Bacterial flora and parasitic fauna of M. leidyi an exotic invader jelly fish to Caspian Sea ecosystem and B. ovata to Black sea an alternative biological control agent was studied. During summer 1382 to spring 1383, using routine Bacteriological work. 72 sample of sea water Caspian Sea obtained from depth 20 and 50 meters, 36 sample of M. leidyi from depth 20 meters, 10 sample of B. ovata and 3 sample of sea water (Black sea) were collected and according to Bacteriological was studied. 216 sample of M. leidyi from depth 10 to 50 meters of Caspian Sea and 47 sample of B. ovata from Black and Marmarreh Sea (Turkey) were collected and was studied. In this study no parasite from was identified in M. leidyi (Caspian sea) but 64 percentage and 73 percentage of B. ovata (Marmarreh and Black sea respectively) contaminated to Trichodina ctenophore at varians concentration B. ovata of Black sea (130 min 1050 max) and B. ovata Marmarre sea (420 min 2100 max). While B. ovata kept at high salinity of 21 ppt was more contaminated with this pretrichial protozoan (Trichodina) than in low salinity (12/5 ppt). in comparision of bacterial flore in two cetenophore (M. leidyi and B. ovata) was observed that some of bacteria such as micrococcus sp, Aeromonas sp. Bacillus coagulans in both ctenophore and some other bacteria such as Agromobacterium and chromobacterium only observed in B. ovata but other researcher have reported fram Caspin sea and some of bacteria to specific Shewanella , Vibrio harveiy and bacillus linens was observed in B. ovata . Of course specific bacteria cannot transfer to Caspian Sea (different of salinity black sea (2/1%) to Caspian Sea (1/25 %)). Therefore if B. ovata to introduce to south Caspian Sea for biological control population M. leidyi. it is necessary at first some of viral pathogen in aquatic animal (fish) such as VNN, IPN,IHN,VHS,SVC was studied and then with confidence 95% non-infestation B. ovata to viruses and pass from bath anti parasite and anti-bacterial must be introduce to south Caspian sea