Search results for: screening

Authors: Mitra Savabi-Esfahani, Fariba Taleghani, Mahnaz Noroozi, Maryam Tabatabaeian, Elsebeth Lynge

Abstract:

Breast cancer is a health concern in worldwide. All women have not adequate information about breast cancer, resulting in undetected some tumors until advanced stages. Therefore awareness of people was recommended as a strategy to control that. The aim of this study was to assess the effect of an educational intervention on women's knowledge and attitude about breast cancer and screening. This study was conducted in 2016 on 191 women. All women living in one of big cities were invited to enroll in training classes. Inclusion criteria consisted women who were 20 - 69 years and not participated in any educational intervention. The lecture with group discussion was used as educational methods. Data collection tool was a structured questionnaire which filled out before and after intervention. The reliability of the questionnaire was determined by Cronbach's alpha. The data were analyzed using SPSS software. The average age was 44/4 ± 11.5 and 42.6% of the women had obtained high school. Of the 191 women, 70(36.6%) and 76(39.8%) had low and medium level of knowledge respectively and half of them, 95(50%) had medium level of attitude in before intervention. There was significant difference between mean scores of knowledge and attitude before and after the intervention by Paired T test (p < 0/001). It seems applying effective educational interventions can increase knowledge and attitude women about breast cancer particularly in community that they have insufficient levels. Moreover, the lecture method along with group discussion can be proposed as effective and conventional methods for this purpose.

Keywords: attitude, breast cancer, educational intervention, knowledge

Procedia PDF Downloads 286

Authors: Abdul Musaweer Habib, Habibul Hasan Mazumder, Saiful Islam, Sohel Sikder, Omar Faruk Sikder

Abstract:

The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1499 proteins of Fusobacterium nucleatum, which has no homolog in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the KEGG Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the 3-D structure of these three proteins. Finally, determination of ligand binding sites of the key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against Fusobacterium nucleatum.

Keywords: colorectal cancer, drug target, Fusobacterium nucleatum, homology modeling, ligands

Procedia PDF Downloads 362

Authors: Saji George, Eng Khuan Seng, Christof Luda

Abstract:

Nanotechnology has been recognized as an important tool for modern agriculture and has the potential to overcome some of the pressing challenges faced by aquaculture industry. A strategy for optimizing nanotechnology-based therapeutic delivery platform for immunizing farmed fish was developed. Accordingly, a compositional library of nanomaterials of natural chemistry (Halloysite (clay), Chitosan, Hydroxyapatite, Mesoporous Silica and a composite material of clay-chitosan) was screened for their toxicity and efficiency in delivering models antigens in cellular and zebrafish embryo models using high throughput screening platforms. Through multi-parametric optimization, chitosan modified halloysite (clay) nanomaterial was identified as an optimal vaccine delivery platform. Further, studies conducted in juvenile seabass showed the potential of clay-chitosan in delivering outer membrane protein of Tenacibaculum maritimum- TIMA (pathogenic bacteria) to and its efficiency in eliciting immune responses in fish. In short, as exemplified by this work, the strategy of using compositional nanomaterial libraries and their biological profiling using high-throughput screening platform could fasten the discovery process of nanomaterials with potential applications in food and agriculture.

Keywords: nanotechnology, fish-vaccine, drug-delivery, halloysite-chitosan

Procedia PDF Downloads 254

Authors: Sanober Khanum, Barerah Siddiqui, Asim Qidwai

Abstract:

Objective: To determine the risk factors and frequency of depressive trends in children and adolescents suffering from Beta-Thalassemia. Background: Thalassemia is a chronic disease affecting 10,000 people in 60 countries. Many studies show that prolonged medical conditions cause depression. Due to the invasive procedures and suffering, Beta-Thalassemia cause great psychological distress to both children and their caregivers. The study shows 14-24% prevalence of psychiatric problems in Thalassemic patients. Method: Sample consisted of 195 registered patients of A.M.T.F (Female=95 and Male=100). Based on age range the sample was divided into two groups, Group A = children (4-9 years) and Group B = adolescent (10-16 years). A detailed interview with a self-made screening measure was administered on parents to find out the level of depression in patients. Statistics: Chi-square and t-test was applied in order to analyze the data. Results show high prevalence of depression, depression n= 131(66.83%), no depression n=65(33.16%). Analyses reflect that age influences the level of depression Adolescent (71.05%) and Children (64.16%). The analysis also shows a difference in level of depression between both genders. (t=2.975, p < .05). Conclusion: There is a high possibility of developing depressive trend in children affected with Beta Thalassemia; especially females. Therefore, there is a dire need for psychological screening and appropriate treatment in order to improve physical; as well as mental health.

Keywords: childhood depression, chronic illness, psychopathology, Thalassemia

Procedia PDF Downloads 305

Authors: P. Sindhumole, K. Soumya, R. Renjimol

Abstract:

Rice (Oryza sativa L.) is the major staple food crop over the world. It is prone to a number of biotic and abiotic stresses, out of which Bacterial Leaf Blight (BLB), caused by Xanthomonas oryzae pv. oryzae, is the most rampant. Management of this disease through chemicals or any other means is very difficult. The best way to control BLB is by the development of Host Plant Resistance. BLB resistance is not an activity of a single gene but it involves a cluster of more than thirty genes reported. Among these, Xa5 and Xa13 genes are two important ones, which can be diagnosed through marker assisted selection using closely linked molecular markers. During 2014, the first phase of field screening using forty traditional rice genotypes was carried out and twenty resistant symptomless genotypes were identified. Molecular characterisation of these genotypes using RM 122 SSR marker revealed the presence of Xa5 gene in thirteen genotypes. Forty-two traditional rice genotypes were used for the second phase of field screening for BLB resistance. Among these, sixteen resistant genotypes were identified. These genotypes, along with two susceptible check genotypes, were subjected to marker assisted selection for Xa13 gene, using the linked STS marker RG-136. During this process, presence of Xa13 gene could be detected in ten resistant genotypes. In future, these selected genotypes can be directly utilised as donors in Marker assisted breeding programmes for BLB resistance in rice.

Keywords: oryza sativa, SSR, STS, marker, disease, breeding

Procedia PDF Downloads 374

Authors: Uzma Ayaz, Sanam Bashir, Shahid Iqbal Awan, Muhammad Ilyas, Muhammad Fareed Khan

Abstract:

Drought stress directly affects growth along with the productivity of plants by altering plant water status. Sunflower (Helianthus annuus L.), an oilseed crop, is adversely affected by abiotic stresses. The present study was carried out to characterize the genetic variability for seedling and morpho-physiological parameters in different sunflower genotypes under water-stressed conditions. A total of twenty-seven genotypes, including two hybrids, eight advanced lines and seventeen accessions of sunflower (Helianthus annuus L.) were tested against drought stress at Seedling stages by Polyethylene glycol (PEG). Significant means were calculated among traits using analysis of variance (ANOVA) whereas, correlation and principal component analysis also confirmed that germination percentage, root length, shoot length, chlorophyll content, stomatal frequency are positively linked with each other hence, these traits were responsible for most of the variation among genotypes. The cluster analysis results showed that genotypes Ausun, line-3, line-2, and 17578, line-1, line-7, line-6 and 17562 as more diverse among all the genotypes. These most divergent genotypes could be utilized in the development of drought-tolerant inbreed lines which could be subsequently used in future heterosis breeding programs.

Keywords: sunflower, drought, stress, polyethylene- glycol, screening

Procedia PDF Downloads 100

Authors: Scott Ashby, Emily Granger, Mark Connellan

Abstract:

Background: In-hospital management of Out-of-Hospital Cardiac Arrest (OHCA) is complex as the aetiologies are varied. Acute coronary angiography has been shown to improve outcomes for patients with coronary occlusion as the cause; however, these patients are difficult to identify. ECG results may help identify these patients, but the accuracy of this diagnostic test is under debate, and requires further investigation. Methods: Arrest and hospital management information was collated retrospectively for OHCA patients who presented to a single clinical site between 2009 and 2013. Angiography results were then collected and checked for significance with survival to discharge. The presence of a severe lesion (>70%) was then compared to categorised ECG findings, and the accuracy of the test was calculated. Results: 104 patients were included in this study, 44 survived to discharge, 52 died and 8 were transferred to other clinical sites. Angiography appears to significantly correlate with survival to discharge. ECG showed 54.8% sensitivity for detecting the presence of a severe lesion within the group that received angiography. A combined criterion including any ECG pathology showed 100% sensitivity and negative predictive value, however, a low specificity and positive predictive value. Conclusion: In the cohort investigated, ST elevation on ECG is not a sensitive enough screening test to be used to determine whether OHCA patients have coronary stenosis as the likely cause of their arrest, and more investigation into whether screening with a combined ECG criterion, or whether all patients should receive angiography routinely following OHCA is needed.

Keywords: out of hospital cardiac arrest, coronary angiography, resuscitation, emergency medicine

Procedia PDF Downloads 377

Authors: Rim Frikha, Nouha Bouayed, Afifa Sellami, Nozha Chakroun, Salima Daoud, Leila Keskes, Tarek Rebai

Abstract:

Introduction: Recurrent pregnancy loss (RPL) defined as two or more pregnancy losses, is a serious clinical problem. Methylene-tetrahydro-folate-reductase (MTHFR) polymorphisms, commonly the variant C677T is recognized as an inherited thrombophilia which might affect embryonic development and pregnancy success and cause pregnancy complications as RPL. Material and Methods DNA was extracted from peripheral blood samples and PCR-RFLP was performed for the molecular diagnosis of the C677T MTHFR polymorphism among 70 patients (35 couples) with more than 2 fetal losses. Aims and Objective: The aim of this study is to determine the frequency of MTHFR C677T among Tunisian couples with RPL and to critically analyze the available literature on the importance of MTHFR polymorphism testing in the management of RPL. Result and comments: No C677T mutation was detected in the carriers of RPL. This result would be related to sample size and to different criteria (number of abortion), - The association between MTHFR polymorphisms and pregnancy complications has been reported but with controversial results. - A lack of evidence for MTHFR polymorphism testing previously recommended by ACMG (American College of Medical medicine). Our study highlights the importance of screening of MTHFR polymorphism since the real impact of such thrombotic molecular defect on the pregnancy outcome is evident. - Folic supplementation of these patients during pregnancy can prevent such complications and lead to a successful pregnancy outcome.

Keywords: methylenetetrahydrofolate reductase, C677T, recurrent pregnancy loss, genetic testing

Procedia PDF Downloads 285

Authors: Nidhi Chadha, A. K. Tiwari, Marilyn D. Milton, Anil K. Mishra

Abstract:

Translocator protein (18 kDa) TSPO is highly expressed during microglia activation in neuroinflammation. Although a number of PET ligands have been developed for the visualization of activated microglia, one of the advantageous approaches is to develop potential optical imaging (OI) probe. Our study involves computational screening, synthesis and evaluation of TSPO ligand through various imaging modalities namely PET/SPECT/Optical. The initial computational screening involves pharmacophore modeling from the library designing having oxo-benzooxazol-3-yl-N-phenyl-acetamide groups and synthesis for visualization of efficacy of these compounds as multimodal imaging probes. Structure modeling of monomer, Ala147Thr mutated, parallel and anti-parallel TSPO dimers was performed and docking analysis was performed for distinct binding sites. Computational analysis showed pattern of variable binding profile of known diagnostic ligands and NBMP via interactions with conserved residues along with TSPO’s natural polymorphism of Ala147→Thr, which showed alteration in the binding affinity due to considerable changes in tertiary structure. Preliminary in vitro binding studies shows binding affinity in the range of 1-5 nm and selectivity was also certified by blocking studies. In summary, this skeleton was found to be potential probe for TSPO imaging due to ease in synthesis, appropriate lipophilicity and reach to specific region of brain.

Keywords: TSPO, molecular modeling, imaging, docking

Procedia PDF Downloads 436

Authors: Shahid Mahboob, Hafiz Muhammad Ashraf, Salma Sultana, Tayyaba Sultana, Khalid Al-Ghanim, Fahid Al-Misned, Zubair Ahmedd

Abstract:

There is an increasing need for an effective tool to estimate the risks derived from the large number of pollutants released to the environment by human activities. Typical screening procedures are highly invasive or lethal to the fish. Recent studies show that fish scales biochemically respond to a range of contaminants, including toxic metals, organic compounds, and endocrine disruptors. The present study evaluated the effects of the surface water contaminants on Cyprinus carpio in the Ravi River by comparing DNA extracted non-lethally from their scales to DNA extracted from the scales of fish collected from a controlled fish farm. A single, random sampling was conducted. Fish were broadly categorised into three weight categories (W1, W2 and W3). The experimental samples in the W1, W2 and W3 categories had an average DNA concentration (µg/µl) that was lower than the control samples. All control samples had a single DNA band; whereas the experimental samples in W1 fish had 1 to 2 bands, the experimental samples in W2 fish had two bands and the experimental samples in W3 fish had fragmentation in the form of three bands. These bands exhibit the effects of pollution on fish in the Ravi River. On the basis findings of this study, we propose that fish scales can be successfully employed as a new non-lethal tool for the evaluation of the effect of surface water contaminants.

Keywords: fish scales, Cyprinus carpio, heavy metals, non-invasive, DNA fragmentation

Procedia PDF Downloads 385

Authors: K. S. Al-Mugren, A. Yahya, S. Alodah, R. Alharbi, S. H. Almsaid , A. Alqahtani, H. Jaber, A. Basaqer, N. Alajra, N. Almoghati, A. Alsalman, Khalid Alharbi

Abstract:

Context: The production of biofertilizers from naturally occurring microorganisms is an area of research that aims to enhance agricultural practices by utilizing local resources. This research project focuses on isolating and screening indigenous microorganisms with PK-fixing and phosphate solubilizing characteristics from local sources. Research Aim: The aim of this project is to develop a biofertilizer product using indigenous microorganisms and composted agro waste as a carrier. The objective is to enhance crop productivity and soil fertility through the application of biofertilizers. Methodology: The research methodology includes several key steps. Firstly, indigenous microorganisms will be isolated from local resources using the ten-fold serial dilutions technique. Screening assays will be conducted to identify microorganisms with phosphate solubilizing and PK-fixing activities. Agro-waste materials will be collected from local agricultural sources, and composting experiments will be conducted to convert them into organic matter-rich compost. Physicochemical analysis will be performed to assess the composition of the composted agro-waste. Gamma and X-ray irradiation will be used to sterilize the carrier material. The sterilized carrier will be tested for sterility using the ten-fold serial dilutions technique. Finally, selected indigenous microorganisms will be developed into biofertilizer products. Findings: The research aims to find suitable indigenous microorganisms with phosphate solubilizing and PK-fixing characteristics for biofertilizer production. Additionally, the research aims to assess the suitability of composted agro waste as a carrier for biofertilizers. The impact of gamma irradiation sterilization on pathogen elimination will also be investigated. Theoretical Importance: This research contributes to the understanding of utilizing indigenous microorganisms and composted agro waste for biofertilizer production. It expands knowledge on the potential benefits of biofertilizers in enhancing crop productivity and soil fertility. Data Collection and Analysis Procedures: The data collection process involves isolating indigenous microorganisms, conducting screening assays, collecting and composting agro waste, analyzing the physicochemical composition of composted agro waste, and testing carrier sterilization. The analysis procedures include assessing the abilities of indigenous microorganisms, evaluating the composition of composted agro waste, and determining the sterility of the carrier material. Conclusion: The research project aims to develop biofertilizer products using indigenous microorganisms and composted agro waste as a carrier. Through the isolation and screening of indigenous microorganisms, the project aims to enhance crop productivity and soil fertility by utilizing local resources. The research findings will contribute to the understanding of the suitability of composted agro waste as a carrier and the efficacy of gamma irradiation sterilization. The research outcomes will have theoretical importance in the field of biofertilizer production and agricultural practices.

Keywords: biofertilizer, microorganisms, agro waste, nuclear technologies

Procedia PDF Downloads 89

Authors: Azizollah Khodakaram- Tafti, Ali Mohammadi, Ghasem Farjanikish

Abstract:

Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens of cattle worldwide caused by Pestivirus genus, Flaviviridae family.The aim of the present study was to comparison several diagnostic methods and determine the prevalence of BVDV infection for the first time in dairy herds of Fars province, Iran. For initial screening, a total of 400 blood samples were randomly collected from 12 industrial dairy herds and analyzed using reverse transcription (RT)-PCR on the buffy coat. In the second step, blood samples and also ear notch biopsies were collected from 100 cattle of infected farms and tested by antigen capture ELISA (ACE), RT-PCR and immunohistochemistry (IHC). The results of nested RT-PCR (outer primers 0I100/1400R and inner primers BD1/BD2) was successful in 16 out of 400 buffy coat samples (4%) as acute infection in initial screening. Also, 8 out of 100 samples (2%) were positive as persistent infection (PI) by all of the diagnostic tests similarly including RT-PCR, ACE and IHC on buffy coat, serum and skin samples, respectively. Immunoreactivity for bovine BVDV antigen as brown, coarsely to finely granular was observed within the cytoplasm of epithelial cells of epidermis and hair follicles and also subcutaneous stromal cells. These findings confirm the importance of monitoring BVDV infection in cattle of this region and suggest detection and elimination of PI calves for controlling and eradication of this disease.

Keywords: antigen capture ELISA, bovine viral diarrhea virus, immunohistochemistry, RT-PCR, cattle

Procedia PDF Downloads 343

Authors: Amal Saafan, Kareem Al Sofy, Sameh AbdelGhani, Magdy Amin

Abstract:

Background: Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production of β-lactamases is the objective of the study. Methods: One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar, MicroScan and PCR amplification of blaoxa-51like gene. Acinetobacterbaumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem and cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST, and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like, and blaADC-like genes was performed for isolates that showed, at least, two positive results of three for both AmpC and carbapenemases phenotypic screening tests (obvious activity), in addition to GP3 (for comparison). Detection of blaoxa-51like and blaADC-like genes preceded by ISAba1 was also performed. Results: Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2, and GP3, respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively.AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51like and blaADC-like genes, while blaoxa-23like, blaIMP-like andblaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. Only 12 (44.4%) isolates harbored blaoxa-51like and blaADC-like genes preceded by ISAba1. GP3 isolates showed only positive blaoxa-51like and blaADC-like genes. Conclusion: It is not possible to correlate resistance with presence of blaoxa-51like and blaADC-like genes and presence of ISAba1 was immediate as transcriptional promoter. A blaoxa-23like gene played an important role in carbapenem resistance when compared with blaIMP-like and blaVIM-like gene.

Keywords: acinetobacter, beta-lactams, resistance, antimicrobial agents

Procedia PDF Downloads 325

Authors: Amer A. Hasan, Mehri Igci, Ersin Borazan, Rozhgar A. Khailany, Emine Bayraktar, Ahmet Arslan

Abstract:

Gastric cancer (GC) has high morbidity and fatality rate in various countries and is still one of the most frequent and deadly diseases. Novel mitogenic and motogenic Gastrokine1 (GKN1) and Gastrokine 2 (GKN2) genes that are highly expressed in the normal stomach epithelium and plays an important role in maintaining the integrity and homeostasis of stomach mucosal epithelial cells. Significant loss of copy number and mRNA transcript of GKN1 and GKN2 gene expression were frequently observed in all types of gastric cancer. In this study, 47 paired samples that were grouped according to the types of gastric cancer and the clinical characteristics of the patients, including gender and average of age were investigated with gene expression analysis and mutation screening by monetering RT-PCR, SSCP and nucleotide sequencing techniques. Both GKN1 and GKN2 genes were observed significantly reduced found by (Wilcoxon signed rank test; p<0.05). As a result of gene screening, no mutation (no different genotype) was detected. It is considered that gene mutations are not the cause of inactivation of gastrokines. In conclusion, the mRNA expression level of GKN1 and GKN2 genes statistically was decreased regardless the gender, age or cancer type of patients. Reduced of gastrokine genes seems to occur at the initial steps of cancer development. In order to understand the investigation between gastric cancer and diagnostic biomarker; further analysis is necessary.

Keywords: gastric cancer, diagnostic biomarker, nucleotide sequencing, semi-quantitative RT-PCR

Procedia PDF Downloads 457

Authors: Anjani Kumar Tiwari, Neelam Kumari, Anil Mishra

Abstract:

The 18-kDa translocator protein (TSPO) previously called as peripheral benzodiazepine receptor, is proven biomarker for variety of neuroinflammation. TSPO is tryptophane rich five transmembranal protein found on outer mitochondrial membrane of steroid synthesising and immunomodulatory cells. In case of neuronal damage or inflammation the expression level of TSPO get upregulated as an immunomodulatory response. By utilizing Benzoxazolone as a basic scaffold, series of TSPO ligands have been designed followed by their screening through in silico studies. Synthesis has been planned by employing convergent methodology in six high yielding steps. For the synthesized ligands the ‘in vitro’ assay was performed to determine the binding affinity in term of Ki. On ischemic rat brain, autoradiography studies were also carried to check the specificity and affinity of the designed radiolabelled ligand for TSPO.Screening was performed on the basis of GScore of CADD based schrodinger software. All the modified and better prospective compound were successfully carried out and characterized by spectroscopic techniques (FTIR, NMR and HRMS). In vitro binding assay showed best binding affinity Ki = 6.1+ 0.3 for TSPO over central benzodiazepine receptor (CBR) Ki > 200. ARG studies indicated higher uptake of two analogues on the lesion side compared with that on the non-lesion side of ischemic rat brains. Displacement experiments with unlabelled ligand had minimized the difference in uptake between the two sides which indicates the specificity of the ligand towards TSPO receptor.

Keywords: TSPO, PET, imaging, Acetamidobenzoxazolone

Procedia PDF Downloads 123

Authors: Julia R. Beulig, Stefan Ohla, Detlev Belder

Abstract:

In contrast to off-line analytical methods, lab-on-a-chip technology delivers direct information about the observed reaction. Therefore, microfluidic devices make an important scientific contribution, e.g. in the field of synthetic chemistry. Herein, the rapid generation of analytical data can be applied for the optimization of chemical reactions. These microfluidic devices enable a fast change of reaction conditions as well as a resource saving method of operation. In the presented work, we focus on the investigation of multiphase regimes, more specifically on a biphasic microfluidic droplet systems. Here, every single droplet is a reaction container with customized conditions. The biggest challenge is the rapid qualitative and quantitative readout of information as most detection techniques for droplet systems are non-specific, time-consuming or too slow. An exception is the electrospray mass spectrometry (ESI-MS). The combination of a reaction screening platform with a rapid and specific detection method is an important step in droplet-based microfluidics. In this work, we present a novel approach for synthesis optimization on the nanoliter scale with direct ESI-MS detection. The development of a droplet-based microfluidic device, which enables the modification of different parameters while simultaneously monitoring the effect on the reaction within a single run, is shown. By common soft- and photolithographic techniques a polydimethylsiloxane (PDMS) microfluidic chip with different functionalities is developed. As an interface for the MS detection, we use a steel capillary for ESI and improve the spray stability with a Teflon siphon tubing, which is inserted underneath the steel capillary. By optimizing the flow rates, it is possible to screen parameters of various reactions, this is exemplarity shown by a Domino Knoevenagel Hetero-Diels-Alder reaction. Different starting materials, catalyst concentrations and solvent compositions are investigated. Due to the high repetition rate of the droplet production, each set of reaction condition is examined hundreds of times. As a result, of the investigation, we receive possible reagents, the ideal water-methanol ratio of the solvent and the most effective catalyst concentration. The developed system can help to determine important information about the optimal parameters of a reaction within a short time. With this novel tool, we make an important step on the field of combining droplet-based microfluidics with organic reaction screening.

Keywords: droplet, mass spectrometry, microfluidics, organic reaction, screening

Procedia PDF Downloads 275

Authors: Md. Fazlul Karim, Ashik Sharfaraz, Aysha Ferdoushi

Abstract:

Background: Computational medicinal chemistry approaches are used for designing and identifying new drug-like molecules, predicting properties and pharmacological activities, and optimizing lead compounds in drug development. SIRT7, a nicotinamide adenine dinucleotide (NAD+)-dependent deacylase which regulates aging, is an emerging target for cancer therapy with mounting evidence that SIRT7 downregulation plays important roles in reversing cancer phenotypes and suppressing tumor growth. Activation or altered expression of SIRT7 is associated with the progression and invasion of various cancers, including liver, breast, gastric, prostate, and non-small cell lung cancer. Objectives: The goal of this work was to identify potent and selective bioactive candidate inhibitors of SIRT7 by in silico screening of small molecule compounds obtained from Nigella sativa (N. sativa). Methods: SIRT7 structure was retrieved from The Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB), and its active site was identified using CASTp and metaPocket. Molecular docking simulation was performed with PyRx 0.8 virtual screening software. Drug-likeness properties were tested using SwissADME and pkCSM. In silico toxicity was evaluated by Osiris Property Explorer. Bioactivity was predicted by Molinspiration software. Antitumor activity was screened for Prediction of Activity Spectra for Substances (PASS) using Way2Drug web server. Molecular dynamics (MD) simulation was carried out by Desmond v3.6 package. Results: A total of 159 bioactive compounds from the N. Sativa were screened against the SIRT7 enzyme. Five bioactive compounds: chrysin (CID:5281607), pinocembrin (CID:68071), nigellidine (CID:136828302), nigellicine (CID:11402337), and epicatechin (CID:72276) were identified as potent SIRT7 anti-cancer candidates after docking score evaluation and applying Lipinski's Rule of Five. Finally, MD simulation identified Chrysin as the top SIRT7 anti-cancer candidate molecule. Conclusion: Chrysin, which shows a potential inhibitory effect against SIRT7, can act as a possible anti-cancer drug candidate. This inhibitor warrants further evaluation to check its pharmacokinetics and pharmacodynamics properties both in vitro and in vivo.

Keywords: SIRT7, antitumor, molecular docking, molecular dynamics simulation

Procedia PDF Downloads 48

Authors: Ahmad Ali Shahid, M Shakil Shaukat

Abstract:

Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.

Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR

Procedia PDF Downloads 372

Authors: H. Aldewachi, M. Hines, M. McCulloch, N. Woodroofe, P. Gardiner

Abstract:

DPP-IV is an enzyme whose expression is affected in a variety of diseases, therefore, has been identified as possible diagnostic or prognostic marker for various tumours, immunological, inflammatory, neuroendocrine, and viral diseases. Recently, DPP-IV enzyme has been identified as a novel target for type II diabetes treatment where the enzyme is involved. There is, therefore, a need to develop sensitive and specific methods that can be easily deployed for the screening of the enzyme either as a tool for drug screening or disease marker in biological samples. A variety of assays have been introduced for the determination of DPP-IV enzyme activity using chromogenic and fluorogenic substrates, nevertheless these assays either lack the required sensitivity especially in inhibited enzyme samples or displays low water solubility implying difficulty for use in vivo samples in addition to labour and time-consuming sample preparation. In this study, novel strategies based on exploiting the high extinction coefficient of gold nanoparticles (GNPs) are investigated in order to develop fast, specific and reliable enzymatic assay by investigating synthetic peptide sequences containing a DPP IV cleavage site and coupling them to GNPs. The DPP IV could be detected by colorimetric response of peptide capped GNPs (P-GNPS) that could be monitored by a UV-visible spectrophotometer or even naked eyes, and the detection limit could reach 0.01 unit/ml. The P-GNPs, when subjected to DPP IV, showed excellent selectivity compared to other proteins (thrombin and human serum albumin) , which led to prominent colour change. This provided a simple and effective colorimetric sensor for on-site and real-time detection of DPP IV.

Keywords: gold nanoparticles, synthetic peptides, colorimetric detection, DPP-IV enzyme

Procedia PDF Downloads 283

Authors: Oseni Margaret Oladunni, Ogundele Joan Olayinka, Olusanya Olalekan Samuel, Akinniyi Modupe Olakintan

Abstract:

The world's growing population is pushing humans to look for alternative food sources among underutilised or wild plants. One of these food sources has been identified as Canavalia enisiformis, or jack beans. The only issue with using jack beans is that they contain anti-nutrient chemicals, which must be removed or diminished in order for them to be fit for human consumption. The objective of this study is to determine the nutritional and industrial utility of Canavalia enisiformis by analysing the anti-nutrient, phytochemical, and antioxidant composition of raw whole seed and soaking dehusked seeds using established procedures. Phytate (23.48±0.24, 15.24±0.41 and 14.83±0.00), oxalate (4.32±0.09, 3.96±0.09 and 2.88±0.09), tannins (22.77±0.73, 18.68±0.03 and 17.50±0.46), and lectins (6.67±0.04, 6.20±0.01 and 6.42±0.07) exhibited the highest anti-nutrient values in raw whole seed and, at the very least, in dehusked, soaked seeds. The samples were subjected to phytochemical screening, which detected the presence of cardiac glycosides as well as anthraquinones, alkaloids, tannins, saponins, steroids, flavonoids, terpenoids, phlobatannins, and flavonoids. Due to the reduction in phytochemical contents quantified as a result of dehusking and soaking, phenolbatannins and anthraquinones were not found in the samples. The research findings also demonstrated elevated concentrations of several plausible phytochemical components with potential medical value, with the raw whole seed exhibiting the greatest capacity to scavenge free radicals. Accordingly, the study's findings validate the seed's therapeutic applications and imply that it might be an inexpensive source of antioxidants for humans and animals alike.

Keywords: dehusking, soaking, anti-nutrients, antioxidants, jack bean

Procedia PDF Downloads 24

Authors: Z. Razi, S. M. J. Mortazavi, N. Shokrpour, Z. Shayan, F. Amiri

Abstract:

Long-term exposure to low doses of ionizing radiation occurs in radiation health care workplaces. Although doses in health professions are generally very low, there are still matters of concern. The radiation safety program promotes occupational radiation safety through accurate and reliable monitoring of radiation workers in order to effectively manage radiation protection. To achieve this goal, it has become mandatory to implement health examination periodically. As a result, based on the hematological alterations, working populations with a common occupational radiation history are screened. This paper calls into question the effectiveness of blood component analysis as a screening program which is mandatory for medical radiation workers in some countries. This study details the distribution and trends of changes in blood components, including white blood cells (WBCs), red blood cells (RBCs) and platelets as well as received cumulative doses from occupational radiation exposure. This study was conducted among 199 participants and 100 control subjects at the medical imaging departments at the central hospital of Shiraz University of Medical Sciences during the years 2006–2010. Descriptive and analytical statistics, considering the P-value<0.05 as statistically significance was used for data analysis. The results of this study show that there is no significant difference between the radiation workers and controls regarding WBCs and platelet count during 4 years. Also, we have found no statistically significant difference between the two groups with respect to RBCs. Besides, no statistically significant difference was observed with respect to RBCs with regards to gender, which has been analyzed separately because of the lower reference range for normal RBCs levels in women compared to men and. Moreover, the findings confirm that in a separate evaluation between WBCs count and the personnel’s working experience and their annual exposure dose, results showed no linear correlation between the three variables. Since the hematological findings were within the range of control levels, it can be concluded that the radiation dosage (which was not more than 7.58 mSv in this study) had been too small to stimulate any quantifiable change in medical radiation worker’s blood count. Thus, use of more accurate method for screening program based on the working profile of the radiation workers and their accumulated dose is suggested. In addition, complexity of radiation-induced functions and the influence of various factors on blood count alteration should be taken into account.

Keywords: blood cell count, mandatory testing, occupational exposure, radiation

Procedia PDF Downloads 442

Authors: Nadiya Zahra Karimi, Amir Ghiami Rad

Abstract:

The physical health of students from an early age and the proper formation of the musculoskeletal system of their body is part of the overall health of these people. Most chronic musculoskeletal problems and pains can be controlled and reduced with education at an early age. Therefore, with the correct and timely diagnosis of these abnormalities, we can play an important role in their proper treatment and control, and in a way, raise the level of quality of life and positive self-concept in students. The aim of this study was to investigate the relationship between erectile dysfunctions of the upper limbs (head and neck, shoulder, thoracic and lumbar) and the quality of life and self-concept of male students aged 7 to 10 years. The statical population of the study consists of 227 students of shahadat boys’ primary school in khajeh city. Due to the corona pandemic conditions, the research samples were identified after screening and available according to the entrance criteria of the study. To validate the quality of life, the valid WHOQOL-BREF questionnaire will be used for self-concept variables, Dolatabadi, Fatemeh (2007) questionnaire, and for physical screening, a checkerboard, plumb line, and flexible ruler will be used. There is a negative and significant relationship between the dimensions of upper limb anomalies and quality of life factors, and also there is a negative and significant relationship between the dimensions of upper limb anomalies and self-concept factors. The results showed that there is a negative and significant relationship between head and neck abnormalities with quality of life and self-concept factors, with a significance level of less than 0.05 in male students aged 7 to 10 years.

Keywords: upper limb erectile dysfunction, quality of life, self-concept, erectile abnormalities

Procedia PDF Downloads 73

Authors: B. O. Oyedoyin, M. O. Ilori, T. O. Oyebisi, B. A. Oluwale, O. O. Jegede

Abstract:

The study was designed to evaluate the business development and transfer of technologies to small manufacturing companies by research institutes in South Western Nigeria. The study covered all the industrial research institutions with headquarters in South Western Nigeria. The study showed that the involvement of scientists in innovation process was rated highest in the idea generation (4.14) and idea screening (4.29) phases; high in R&D (3.86) and fairly high in pilot plant development (2.71) and commercialization (2.43) phase. Their involvement was rated low in business analysis and development (2.14), and test marketing (2.29) phase. The involvement of engineers was rated highest in idea generation (3.28), fairly high in R&D (2.71), pilot plant development (2.57), and idea screening (2.40) phases. However, their involvement was rated low in business analysis and development (2.0), test marketing (2.0), and commercialization (1.28) phases. The involvement of technology marketers in innovation process was generally rated fairly high in R&D (2.7) and business analysis and development (2.6), and low in all the other phases of innovation. However, their involvement at IAR&T, FIIRO, and NIOMR in all the phases was rated very high (3.0-5.0). The involvement of entrepreneurs was generally rated from fairly high to low (2.7-2.3) in all the phases of innovation. The involvement of financial institutions in all the phases of innovation was generally rated low (1.28-1.71). In conclusion, the study showed that the involvement of stakeholders like entrepreneurs and financial institutions in technology packaging for commercialization is very low.

Keywords: research institutes, national innovation system, Nigeria, entrepreneurs, financial institution

Procedia PDF Downloads 402

Authors: D. Hilton

Abstract:

An analysis of the Australian Diabetes Screening Study estimated undiagnosed diabetes mellitus [DM] prevalence in a high risk general practice based cohort. DM prevalence varied from 9.4% to 18.1% depending upon the diagnostic criteria utilised with age being a highly significant risk factor. Utilising the gold standard oral glucose tolerance test, the prevalence of DM was 22-23% in those aged >= 70 years and <15% in those aged 40-59 years. Opportunistic screening in Australian general practice potentially can identify many persons with undiagnosed type 2 DM. An Australian Bureau of Statistics document published three years ago, reported the highest rate of DM in men aged 65-74 years [19%] whereas the rate for women was highest in those over 75 years [13%]. If you consider that the Australian Bureau of Statistics report in 2007 found that 13% of the population was over 65 years of age and that this will increase to 23-25% by 2056 with a further projected increase to 25-28% by 2101, obviously this information has to be factored into the equation when age related diabetes prevalence predictions are calculated. This 10-15% proportional increase of elderly persons within the population demographics has dramatic implications for the estimated number of elderly persons with DM in these age groupings. Computational methodology showing the age related demographic changes reported in these official statistical documents will be done showing estimates for 2056 and 2101 for different age groups. This has relevance for future diabetes prevalence rates and shows that along with many countries worldwide Australia is facing an increasing pandemic. In contrast Japan is expected to have a decrease in the next twenty years in the number of persons with diabetes.

Keywords: epidemiological methods, aging, prevalence, diabetes mellitus

Procedia PDF Downloads 353

Authors: J. E. O. Hernandez

Abstract:

In the next 20 years, energy production by burning fuels will increase and so will the atmospheric concentration of CO2 and its well-known threats to life on Earth. The technologies available for capturing CO2 are still dubious and this keeps fostering an interest in bio-inspired approaches. The leading one is the application of carbonic anhydrase (CA) –a superfast biocatalyst able to convert up to one million molecules of CO2 into carbonates in water. However, natural CA underperforms when applied to real smoky CO2 in chimneys and, so far, the efforts to create superior CAs in the lab rely on screening methods running under pristine conditions at the micro level, which are far from resembling those in chimneys. For the evolution of man-made enzymes, selection rather than screening would be ideal but this is challenging because of the need for a suitable artificial environment that is also sustainable for our society. Herein we present the stepwise design and construction of a bioprocess (from bench-scale to semi-pilot) for evolutionary selection experiments. In this bioprocess, reaction and adsorption took place simultaneously at atmospheric pressure in a spray tower. The scrubbing solution was fed countercurrently by reusing municipal pressure and it was mainly prepared with water, carbonic anhydrase and calcium chloride. This bioprocess allowed for the enzymatic carbonation of smoky CO2; the reuse of process water and the recovery of solid carbonates without cooling of smoke, pretreatments, solvent amines and compression of CO2. The average yield of solid carbonates was 0.54 g min-1 or 12-fold the amount produced in serum bottles at lab bench scale. This bioprocess could be used as a tailor-made environment for driving the selection of superior CAs. The bioprocess and its match CA could be sustainably used to reduce global warming by CO2 emissions from exhausts.

Keywords: biological carbon capture and sequestration, carbonic anhydrase, directed evolution, global warming

Procedia PDF Downloads 175

Authors: Ahmed Samy Elnoby

Abstract:

Ailanthus altissima native to Asia which belongs to the family Simaroubaceae was subjected to phytochemical screening and biological investigations. Phytochemical screening revealed the presence of carbohydrates, tannins, sterols, flavonoids and traces of saponins. In addition, quantitative determination of phenolics and flavonoid content were performed. The antimicrobial activity of methanolic extract of the leaves was determined against gram-positive, gram-negative bacteria in addition to fungi using a modified Kirby-Bauer disc diffusion method that was compared with standard discs ampicillin which acts as an antibacterial agent and amphotericin B which acts as an antifungal agent. A high potency was observed against gram-positive bacteria mainly staphylococcus aureus, gram-negative bacteria mainly Escherichia coli and showed no potency against fungi mainly Aspergillus flavus and candida albicans. On the other hand, the antioxidant activity of the extract was determined by 1, 1-diphenyl-2- diphenyl-2-picryl-hydrazil (DPPH). A very low potency was shown by using DPPH for the antioxidant effect so IC50 = 0 ug/ml, IC90 =0 ug /ml and remark gave 47.2 % at 100 ug/ml which is very weak. Cytotoxic activity was determined by using MTT assay (3-4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) against MCF7 (Human Caucasian breast adenocarcinoma) cell line. A moderate potency was shown by using MCF7 cell line for cytotoxic effect so LC50= 90.2 ug/ml, LC90=139.9 ug/ml and the remark gave 55.2% at 100 ug/ml which is of moderate activity so, Ailanthus altissima can be considered to be a promising antimicrobial agent from natural origin.

Keywords: Ailanthus altissima, TLC, HPLC, anti-microbial activity, antifungal activity, antioxidant, cytotoxic activity

Procedia PDF Downloads 154

Authors: Muhammad Solikhudin Nafi, Wahyu Afif Mufida, Mita Erna Wati, Fitri Setyani Rokim, M. Al-Rizqi Dharma Fauzi

Abstract:

High dose activity without any pre-exercise will impact Delayed Onset Muscle Soreness (DOMS). DOMS known as delayed pain post-exercise and induce skeletal injury which will decrease athletes’ performances. From now on, post-exercise muscle damage can be detected by measuring skeletal troponin I (sTnI) concentration in serum using ELISA but this method needs more time and cost. To prevent decreased athletes performances, screening need to be done rapidly. We want to introduce our new prototype to detect DOMS acutely. Rapid detection tests are based on immunological reaction between skeletal troponin I antibodies and sTnI in human serum or whole blood. Chemical methods that are used in the manufacture of diagnostic test is lateral flow immunoassay. The material used is rat monoclonal antibody sTnI, colloidal gold, anti-mouse IgG, nitrocellulose membrane, conjugate pad, sample pad, wick and backing card. The procedure are made conjugate (colloidal gold and mAb sTnI) and insert into the conjugate pad, gives spray sTnI mAb and anti-mouse IgG into nitrocellulose membrane, and assemble RDT. RDT had been evaluated by measuring the sensitivity of positive human serum (n = 30) and negative human serum (n = 30). Overall sensitivity value was 93% and specificity value was 90%. ADCOR as the first rapid detection test qualitatively showed antigen-antibody reaction and showed good overall performances for screening of muscle damage. Furthermore, these finding still need more improvements to get best results.

Keywords: DOMS, sTnI, rapid detection test, ELISA

Procedia PDF Downloads 493

Authors: Hajir, B. Abdllha, , Alaa, I. Mohamed, Khansa, A. Almoniem, Naga, I. Adam, Wdeea, Alhaadi, Ahmed, A. Elshikh, Ahmed, J. Ali, Ismail, G. Makuar, Anas, M. Elnazeer, Nagat, A. Elrofaei, Samir, F. Abdoelftah, Monier, N. Hemidan

Abstract:

The present study was designed to investigate antimicrobial, and antioxidant activities of five species from Acacia (Acacia albidia, Acacia mellifera, Acacia nubica, Acacia seyal var.seyal and Acacia tortilis). Phytochemical study was piloted to detect the bioactive compounds, which have been responsible from the biological activities. The ethanol, chloroform and acetone plant extracts were seasoned against standard bacteria strains of gram +ve bacteria Staphylococcus aureus (ATCC 25923) ,Gram -ve bacteria Pseudomonas aeruginosa (ATCC 27853) and standard fungi Candida albicans (ATCC 90028), using cup-plate method. The antioxidant activities were conducted via DPPH radical scavenging and metal chelating assays. Prospective activity against the five species was observed in acetone extract. Ethanol extract showed highest activities against Staphylococcus aureus, and Candida albicans. Potential antioxidant activity was presented by ethanol. Cholorophorm and acetone extracts via DPPH, the radical scavenging activities were found to be 91±0.03, 88±0.01 and 85±0.04 respectively. The results of phytochemical screening showed that all extracts of studied plant contain flavonoids, saponins, terpenoids, steroids, alkaloids, phenols and tannins. This study give rise to antioxidant, antimicrobial properties of studied plant, and showed interesting correlation with the phytochemical constituents and biological activities.

Keywords: antimicrobial, Antioxidant, Acacia albidia, Acacia mellifera, acacia nubica, acacia seyal var.seyal, Acacia tortilis

Procedia PDF Downloads 535

Authors: Md. Z. Alam, Amal A. Elgharbawy, Muhammad Moniruzzaman, Nassereldeen A. Kabbashi, Parveen Jamal

Abstract:

The enzymatic hydrolysis of lignocellulosic biomass is one of the obstacles in the process of sugar production, due to the presence of lignin that protects the cellulose molecules against cellulases. Although the pretreatment of lignocellulose in ionic liquid (IL) system has been receiving a lot of interest; however, it requires IL removal with an anti-solvent in order to proceed with the enzymatic hydrolysis. At this point, introducing a compatible cellulase enzyme seems more efficient in this process. A cellulase enzyme that was produced by Trichoderma reesei on palm kernel cake (PKC) exhibited a promising stability in several ILs. The enzyme called PKC-Cel was tested for its optimum pH and temperature as well as its molecular weight. One among evaluated ILs, 1,3-diethylimidazolium dimethyl phosphate [DEMIM] DMP was applied in this study. Evaluation of six factors was executed in Stat-Ease Design Expert V.9, definitive screening design, which are IL/ buffer ratio, temperature, hydrolysis retention time, biomass loading, cellulase loading and empty fruit bunches (EFB) particle size. According to the obtained data, IL-enzyme system shows the highest sugar concentration at 70 °C, 27 hours, 10% IL-buffer, 35% biomass loading, 60 Units/g cellulase and 200 μm particle size. As concluded from the obtained data, not only the PKC-Cel was stable in the presence of the IL, also it was actually stable at a higher temperature than its optimum one. The reducing sugar obtained was 53.468±4.58 g/L which was equivalent to 0.3055 g reducing sugar/g EFB. This approach opens an insight for more studies in order to understand the actual effect of ILs on cellulases and their interactions in the aqueous system. It could also benefit in an efficient production of bioethanol from lignocellulosic biomass.

Keywords: cellulase, hydrolysis, lignocellulose, pretreatment

Procedia PDF Downloads 341

Authors: Svetlana Zhikrivetskaya, Nataliya Shirokova, Roman Bikanov, Elizaveta Musatova, Yana Kovaleva, Nataliya Vetrova, Ekaterina Pomerantseva

Abstract:

Nowadays there is a growing number of couples with conceiving problems due to male or female infertility. Genetic abnormalities are responsible for about 31% of all cases of male infertility. These abnormalities include both chromosomal aberrations or aneuploidies and mutations in certain genes. Chromosomal abnormalities can be easily identified, thus the development of screening panels able to reveal genetic reasons of male infertility on gene level is of current interest. There are approximately 2,000 genes involved in male fertility that is the reason why it is very important to determine the most clinically relevant in certain population and ethnic conditions. An infertility screening panel containing 48 mutations in genes AMHR2, CFTR, DNAI1, HFE, KAL1, TSSK2 and AZF locus which are the most clinically relevant for the European population according to databases NCBI and ClinVar was designed. The aim of this research was to confirm clinic relevance of these mutations in the Russian population. Genotyping was performed in 220 patients with different types of male infertility and in 57 healthy males with normozoospermia. Mutations were identified by end-point PCR with TaqMan probes in microfluidic plates. The frequency of 5 mutations in healthy males and 13 mutations in patients with infertility was revealed and estimated. The frequency of mutation c.187C>G in HFE gene was significantly lower for healthy males (8.8%) compared with patients (17.7%) and the values for the European population according to ExAc database (13.7%) and dbSNP (17.2%). Analysis of c.3454G>C, and c.1545_1546delTA mutations in the CFTR gene revealed increased frequency (0.9 and 0.2%, respectively) in patients with infertility compared with data for the European population (0.04%, respectively (ExAc, European (Non-Finnish) and for the Aggregated Populations (0.002% (ExAc), because there is no data for European population for c.1545_1546delTA mutation. The frequency of del508 mutation (CFTR) in patients (1.59%) were lower comparing with male infertility Europeans (3.34-6.25% depending on nationality) and at the same level with healthy Europeans (1.06%, ExAc, European (Non-Finnish). Analysis of c.845G>A (HFE) mutation resulted in decreased frequency in patients (1.8%) in contrast with the European population data (5.1%, respectively, ExAc, European (Non-Finnish). Moreover, obtained data revealed no statistically significant frequency difference for c.845G>A mutation (HFE) between healthy males in the Russian and the European populations. Allele frequencies of mutations c.350G>A (CFTR), c.193A>T (HFE), c.774C>T, and c.80A>G (gene TSSK2) showed no significantly difference among patients with infertility, healthy males and Europeans. Analysis of AZF locus revealed increased frequency for AZFc microdeletion in patients with male infertility. Thereby, the new data of the allele frequencies in infertility patients in the Russian population was obtained. As well as the frequency differences of mutations associated with male infertility among patients, healthy males in the Russian population and the European one were estimated. The revealed differences showed that for high effectiveness of screening panel detecting genetically caused male infertility it is very important to consider ethnic and population characteristics of patients which will be screened.

Keywords: allele frequency, azoospermia, male infertility, mutation, population

Procedia PDF Downloads 372