Search results for: gene delivery
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3336

Search results for: gene delivery

3216 Disruption of MoNUC1 Gene Mediates Conidiation in Magnaporthe oryzae

Authors: Irshad Ali Khan, Jian-Ping Lu, Xiao-Hong Liu, Fu-Cheng Lin

Abstract:

This study reports the functional analysis of a gene MoNUC1 in M. oryzae, which is homologous to the Saccharomyces cerevisiae NUC1 encoding a mitochondrial nuclease protein. The MoNUC1 having a gene locus MGG_05324 is 1002-bp in length and encodes an identical protein of 333 amino acids. We disrupted the gene through gene disruption strategy and isolated two mutants confirmed by southern blotting. The deleted mutants were then used for phenotypic studies and their phenotypes were compared to those of the Guy-11 strain. The mutants were first grown on CM medium to find the effect of MoNUC1 gene disruption on colony growth and the mutants were found to show normal culture colony growth similar to that of the Guy-11 strain. Conidial germination and appressorial formation were also similar in both the mutants and Guy-11 strains showing that this gene plays no significant role in these phenotypes. For pathogenicity, the mutants and Guy-11 mycelium blocks were inoculated on blast susceptible barley seedlings and it was found that both the strains exhibited full pathogenicity showing coalesced and necrotic blast lesions suggesting that this gene is not involved in pathogenicity. Mating of the mutants with 2539 strain formed numerous perithecia showing that MoNUC1 is not essential for sexual reproduction in M. oryzae. However, the mutants were found to form reduced conidia (1.06±8.03B and 1.08±9.80B) than those of the Guy-11 strain (1.46±10.61A) and we conclude that this protein is not required for the blast fungus to cause pathogenicity but plays significant role in conidiation. Proteins of signal transduction pathways that could be disrupted/ intervened genetically or chemically could lead to antifungal products of important fungal cereal diseases and reduce rice yield losses. Tipping the balance toward understanding the whole of pathogenesis, rather than simply conidiation will take some time, but clearly presents the most exciting challenge of all.

Keywords: appressorium formation, conidiation, NUC1, Magnaporthe oryzae, pathogenicity

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3215 Functional Analysis of Thyroid Peroxidase (TPO) Gene Mutations Detected in Patients with Thyroid Dyshormonogenesis

Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

Abstract:

Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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3214 Functional Analysis of Thyroid Peroxidase Gene Mutations Detected in Patients with Thyroid Dyshormonogenesis

Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

Abstract:

Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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3213 A Deletion in Duchenne Muscular Dystrophy Gene Found Through Whole Exome Sequencing in Iran

Authors: Negin Parsamanesh, Saman Ameri-Mahabadi, Ali Nikfar, Mojdeh Mansouri, Hossein Chiti, Gita Fatemi Abhari

Abstract:

Duchenne muscular dystrophy (DMD) is a severe progressive X-linked neuromuscular illness that affects movement through mutations in dystrophin gene. The mutation leads to insufficient, lack of or dysfunction of dystrophin. The cause of DMD was determined in an Iranian family. Exome sequencing was carried out along with a complete physical examination of the family. In silico methods were applied to find the alteration in the protein structure. The homozygous variant in DMD gene (NM-004006.2) was defined as c.2732-2733delTT (p.Phe911CysfsX8) in exon 21. In addition, phylogenetic conservation study of the human dystrophin protein sequence revealed that phenylalanine 911 is one of the evolutionarily conserved amino acids. In conclusion, our study indicated a new deletion in the DMD gene in the affected family. This deletion with an X-linked inheritance pattern is new in Iran. These findings could facilitate genetic counseling for this family and other patients in the future.

Keywords: duchenne muscular dystrophy, whole exome sequencing, iran, metabolic syndrome

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3212 Ocular Delivery of Charged Drugs Using Iontophoresis

Authors: Abraham J. Domb

Abstract:

Nearly every eye disorder and treatment of post operated eyes evolve around ocular drug delivery. Most ocular diseases are treated with repeated topical applications administered as eye drops. Various attempts have been made to improve drug bioavailability by increasing both the retention of the drug in the pre-corneal area and the penetration of the drug through the cornea. However, currently marketed products are associated with vision blurring, irritability, patient discomfort, toxicity, low drug bioavailability, manufacturing difficulties and inadequate aqueous stability. It has been suggested to use iontophoresis for the non-invasive delivery of drugs. The iontophoretic device is composed of a control panel, two electrodes, a cylindrical well for the insertion of a disposable hydrogel, and a disposable hydrogel pellet. The drug-loaded hydrogel is attached to a cylindrical well at the edge of the electrode of the device and placed onto the eye. The device applies a variable electrical current that can vary from 0.1 mA to 1.5 mA for pre-set periods from 10 seconds to 300 seconds. The iontophoretic device developed in the lab was found to be effective in the delivery of the drugs: gentamicin, water-soluble steroids, and various anticancer agents. When testing in rabbits for safety, the device was considered to be non-toxic and effective.

Keywords: iontophoresis, eye disorder, drug delivery, hydrogel

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3211 The Interrelationship between Formal and Informal Institutions and Its Impacts on the Autonomy of Public Service Delivery Units: The Case of Vietnam

Authors: Minh Thi Hai Vo

Abstract:

This article draws on in-depth interviews with state employees at public hospitals and universities in its institutional analysis of the autonomy practices of public service delivery units in Vietnam. Unlike many empirical and theoretical studies that view formal and informal institutions as complements or substitutes, this article finds no evidence of complementary or substitutive relationships. Instead, the article finds that formal institutions accommodate informal ones and that informal institutions tend to compete and interfere, with the existing and ineffective formal institutions. The result of such conflicting relationship is that the actual autonomy of public service delivery units is, in most cases, perceived to be greater than the formal autonomy they are given. In the condition of poor regulation, the informal autonomy may result in unethical practices including rent-seeking and corruption. The implication of the study finding is policy-makers need to redesign and reorganize the autonomisation of public service delivery units to make informal institutions support and reinforce formal ones in a complementary manner.

Keywords: autonomy, formal institutions, informal institutions, public service delivery units, Vietnam

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3210 Delivery of Positively Charged Proteins Using Hyaluronic Acid Microgels

Authors: Elaheh Jooybar, Mohammad J. Abdekhodaie, Marcel Karperien, Pieter J. Dijkstra

Abstract:

In this study, hyaluronic acid (HA) microgels were developed for the goal of protein delivery. First, a hyaluronic acid-tyramine conjugate (HA-TA) was synthesized with a degree of substitution of 13 TA moieties per 100 disaccharide units. Then, HA-TA microdroplets were produced using a water in oil emulsion method and crosslinked in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). Loading capacity and the release kinetics of lysozyme and BSA, as model proteins, were investigated. It was shown that lysozyme, a cationic protein, can be incorporated efficiently in the HA microgels, while the loading efficiency for BSA, as a negatively charged protein, is low. The release profile of lysozyme showed a sustained release over a period of one month. The results demonstrated that the HA-TA microgels are a good carrier for spatial delivery of cationic proteins for biomedical applications.

Keywords: microgel, inverse emulsion, protein delivery, hyaluronic acid, crosslinking

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3209 Ribosomal Protein S4 Gene: Exploring the Presence in Syrian Strain of Leishmania Tropica Genome, Sequencing it and Evaluating Immune Response of pCI-S4 DNA Vaccine

Authors: Alyaa Abdlwahab

Abstract:

Cutaneous leishmaniasis represents a serious health problem in Syria; this problem has become noticeably aggravated after the civil war in the country. Leishmania tropica parasite is the main cause of cutaneous leishmaniasis in Syria. In order to control the disease, we need an effective vaccine against leishmania parasite. DNA vaccination remains one of the favorable approaches that have been used to face cutaneous leishmaniasis. Ribosomal protein S4 is responsible for important roles in Leishmania parasite life. DNA vaccine based on S4 gene has been used against infections by many species of Leishmania parasite but leishmania tropica parasite, so this gene represents a good candidate for DNA vaccine construction. After proving the existence of ribosomal protein S4 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), sequencing it and cloning it into pCI plasmid, BALB/C mice were inoculated with pCI-S4 DNA vaccine. The immune response was determined by monitoring the lesion progression in inoculated BALB/C mice for six weeks after challenging mice with Leishmania tropica (LCED Syrian 01) parasites. IL-12, IFN-γ, and IL-4 were quantified in draining lymph nodes (DLNa) of the immunized BALB/C mice by using the RT-qPCR technique. The parasite burden was calculated in the final week for the footpad lesion and the DLNs of the mice. This study proved the existence and the expression of the ribosomal protein S4 gene in Leishmania tropica (LCED Syrian 01) promastigotes. The sequence of ribosomal protein cDNA S4 gene was determined and published in Genbank; the gene size was 822 bp. Expression was also demonstrated at the level of cDNA. Also, this study revealed that pCI-S4 DNA vaccine induces TH1\TH2 response in immunized mice; this response prevents partially developing a dermal lesion of Leishmania.

Keywords: ribosomal protein S4, DNA vaccine, Leishmania tropica, BALB\c

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3208 Application of Digital Technologies as Tools for Transformative Agricultural Science Instructional Delivery in Secondary Schools

Authors: Cajethan U. Ugwuoke

Abstract:

Agriculture is taught in secondary schools to develop skills in students which will empower them to contribute to national economic development. Unfortunately, our educational system emphasizes the application of conventional teaching methods in delivering instructions, which fails to produce students competent enough to carry out agricultural production. This study was therefore aimed at examining the application of digital technologies as tools for transformative instructional delivery. Four specific purposes, research questions and hypotheses guided the study. The study adopted a descriptive survey research design where 80 subjects representing 64 teachers of agriculture and 16 principals in the Udenu local government area of Enugu State, Nigeria, participated in the study. A structured questionnaire was used to collect data. The assumption of normality was ascertained by subjecting the data collected to a normality test. Data collected were later subjected to mean, Pearson product-moment correlation, ANOVA and t-test to answer the research questions and test the hypotheses at a 5% significant level. The result shows that the application of digital technologies helps to reduce learners’ boredom (3.52.75), improves learners’ performance (3.63.51), and is used as a visual aid for learners (3.56.61), among others. There was a positive, strong and significant relationship between the application of digital technologies and effective instructional delivery (+.895, p=.001<.05, F=17.73), competency of teachers to the application of digital technologies and effective instructional delivery (+998, p=.001<0.5, F=16263.45), and frequency of the application of digital technologies and effective instructional delivery (+.999, p=.001<.05, F=31436.14). There was no evidence of autocorrelation and multicollinearity in the regression models between the application of digital technologies and effective instructional delivery (2.03, Tolerance=1.00, VIF=1.00), competency of teachers in the application of digital technologies and effective instructional delivery (2.38, Tolerance=1.00, VIF=1.00) and frequency of the application of digital technologies and effective instructional delivery (2.00, Tolerance=1.00, VIF=1.00). Digital technologies should be therefore applied in teaching to facilitate effective instructional delivery in agriculture.

Keywords: agricultural science, digital technologies, instructional delivery, learning

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3207 Repositioning Nigerian University Libraries for Effective Information Provision and Delivery in This Age of Globalization

Authors: S. O. Uwaifo

Abstract:

The paper examines the pivotal role of the library in university education through the provision of a wide range of information materials (print and non- print) required for the teaching, learning and research activities of the university. However certain impediments to the effectiveness of Nigerian university libraries, such as financial constraints, high foreign exchange, global disparities in accessing the internet, lack of local area networks, erratic electric power supply, absence of ICT literacy, poor maintenance culture, etc., were identified. Also, the necessity of repositioning Nigerian university libraries for effective information provision and delivery was stressed by pointing out their dividends, such as users’ access to Directory of Open Access Journals (DOAJ), Online Public Access Catalogue (OPAC), Institutional Repositories, Electronic Document Delivery, Social Media Networks, etc. It therefore becomes necessary for the libraries to be repositioned by way of being adequately automated or digitized for effective service delivery, in this age of globalization. Based on the identified barriers by this paper, some recommendations were proffered.

Keywords: repositioning, Nigerian university libraries, effective information provision and delivery, globalization

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3206 Effect of SCN5A Gene Mutation in Endocardial Cell

Authors: Helan Satish, M. Ramasubba Reddy

Abstract:

The simulation of an endocardial cell for gene mutation in the cardiac sodium ion channel NaV1.5, encoded by SCN5A gene, is discussed. The characterization of Brugada Syndrome by loss of function effect on SCN5A mutation due to L812Q mutant present in the DII-S4 transmembrane region of the NaV1.5 channel protein and its effect in an endocardial cell is studied. Ten Tusscher model of human ventricular action potential is modified to incorporate the changes contributed by L812Q mutant in the endocardial cells. Results show that BrS-associated SCN5A mutation causes reduction in the inward sodium current by modifications in the channel gating dynamics such as delayed activation, enhanced inactivation, and slowed recovery from inactivation in the endocardial cell. A decrease in the inward sodium current was also observed, which affects depolarization phase (Phase 0) that leads to reduction in the spike amplitude of the cardiac action potential.

Keywords: SCN5A gene mutation, sodium channel, Brugada syndrome, cardiac arrhythmia, action potential

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3205 Service Delivery Process in the Luxury Hotel Industry in Dubai: A Hoteliers’ Perspective

Authors: Veronique Gregorec, Prakash Vel, Collins A. Brobbey

Abstract:

Service delivery process in the face of ever changing customer expectations could not have been more important in glamorous Dubai luxury hotel service sector. Based on in-depth discussions with Dubai luxury hotel service pioneers, customer expectations, service processes, customer complaining behavior, and service recovery strategies in the luxury hotel industry are evaluated from the perspectives of service providers. Findings are in agreement with the statement that in the service industry the customer is not always right, and that hotel service providers have acknowledged the need to take extra measures towards individualized and personal service experience delivery. Ultimately, hoteliers set highest standards at all stages of the service delivery process in order to achieve positive and high customer ratings in all customer evaluation areas.

Keywords: luxury hotels, Dubai hotels, Dubai hospitality industry, guest service process

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3204 DNA Barcoding of Tree Endemic Campanula Species From Artvi̇n, Türki̇ye

Authors: Hayal Akyildirim Beğen, Özgür Emi̇nağaoğlu

Abstract:

DNA barcoding is the method of description of species based on gene diversity. In current studies, registration, genetic identification and protection of especially endemic plants pecies are carried out by DNA barcoding techniques. Molecular studies are based on the amplification and sequencing of the barcode gene region by the PCR method. Endemic Campanula choruhensis Kit Tan & Sorger, Campanula troegera Damboldt and Campanula betulifolia K.Koch is widespread in Artvin, Erzurum and around Çoruh valley passing through it. Intense road and dam constructions are carried out in and around the distribution area of this species. This situation harms the habitat of the species and puts its extinction. In this study, the plastid matK barcode gene regions (650 bp) of three Campanula species were created. To make the identification of this species quickly and accurately, gene sequence compared with sequences of other Campanula L. species. As a result of phylogenetic analysis, C. choruhensis is close relative to C. betulifolia. Morphologically, these species were determined to be more similar to each other with flower and leaf characters. C. troegera formed a separate branch.

Keywords: campanula, DNA barcoding, endemic, türkiye, artvin

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3203 Cas9-Assisted Direct Cloning and Refactoring of a Silent Biosynthetic Gene Cluster

Authors: Peng Hou

Abstract:

Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

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3202 Implementing Delivery Drones in Logistics Business Process: Case of Pharmaceutical Industry

Authors: Nikola Vlahovic, Blazenka Knezevic, Petra Batalic

Abstract:

In this paper, we will present a research about feasibility of implementing unmanned aerial vehicles, also known as 'drones', in logistics. Research is based on available information about current incentives and experiments in application of delivery drones in commercial use. Overview of current pilot projects and literature, as well as an overview of detected challenges, will be compiled and presented. Based on these findings, we will present a conceptual model of business process that implements delivery drones in business to business logistic operations. Business scenario is based on a pharmaceutical supply chain. Simulation modeling will be used to create models for running experiments and collecting performance data. Comparative study of the presented conceptual model will be given. The work will outline the main advantages and disadvantages of implementing unmanned aerial vehicles in delivery services as a supplementary distribution channel along the supply chain.

Keywords: business process, delivery drones, logistics, simulation modelling, unmanned aerial vehicles

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3201 Critical Factors Influencing Effective Communication Among Stakeholders on Construction Project Delivery in Jigawa State, Nigeria

Authors: Shazali Abdulahi

Abstract:

Project planning is the first phase in project life cycle which relates to the use of schedules such as Gantt charts to plan and subsequently report the project progress within the project environment. Likewise, project execution is the third phase in project lifecycle, is the phase where the work of the project must get done correctly and it’s the longest phase in the project lifecycle therefore, they must be effectively communicated, now today Communication has become the crucial element of every organization. During construction project delivery, information needs to be accurately and timely communicating among project stakeholders in order to realize the project objective. Effective communication among stakeholders during construction project delivery is one of the major factors that impact construction project delivery. Therefore, the aim of the research work is to examine the critical factors influencing effective communication among stakeholders on construction project delivery from the perspective of construction professionals (Architects, Builders, Quantity surveyors, and Civil engineers). A quantitative approach was adopted. This entailed the used of structured questionnaire to one (108) construction professionals in public and private organization within dutse metropolis. Frequency, mean, ranking and multiple linear regression using SPSS vision 25 software were used to analyses the data. The results show that Leadership, Trust, Communication tools, Communication skills, Stakeholders involvement, Cultural differences, and Communication technology were the most critical factors influencing effective communication among stakeholders on construction project delivery. The hypothesis revealed that, effective communication among stakeholders has significant effects on construction project delivery. This research work will profit the construction stakeholders in construction industry, by providing adequate knowledge regarding the factors influencing effective communication among stakeholders, so that necessary steps to be taken to improve project performance. Also, it will provide knowledge about the appropriate strategies to employ in order to improve communication among stakeholders.

Keywords: effetive communication, ineffective communication, stakeholders, project delivery

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3200 Identification of a Novel Maize Dehydration-Responsive Gene with a Potential Role in Improving Maize Drought Tolerance

Authors: Kyle Phillips, Ndiko Ludidi

Abstract:

Global climate change has resulted in altered rainfall patterns, which has resulted in annual losses in maize crop yields due to drought. Therefore it is important to produce maize cultivars that are more drought-tolerant, which is not an easily accomplished task as plants have a plethora of physical and biochemical adaptation methods. One such mechanism is the drought-induced expression of enzymatic and non-enzymatic proteins which assist plants to resist the effects of drought on their growth and development. One of these proteins is AtRD22 which has been identified in Arabidopsis thaliana. Using an in silico approach, a maize protein with 48% sequence homology to AtRD22 has been identified. This protein appears to be localized in the extracellular matrix, similarly to AtRD22. Promoter analysis of the encoding gene reveals cis-acting elements suggestive of induction of the gene’s expression by abscisic acid (ABA). Semi-quantitative transcriptomic analysis of the putative maize RD22 has revealed an increase in transcript levels after the exposure to drought. Current work elucidates the effect of up-regulation and silencing of the maize RD22 gene on the tolerance of maize to drought. The potential role of the maize RD22 gene in maize drought tolerance can be used as a tool to improve food security.

Keywords: abscisic acid, drought-responsive cis-acting elements, maize drought tolerance, RD22

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3199 Identification and Validation of Co-Dominant Markers for Selection of the CO-4 Anthracnose Disease Resistance Gene in Common Bean Cultivar G2333

Authors: Annet Namusoke, Annet Namayanja, Peter Wasswa, Shakirah Nampijja

Abstract:

Common bean cultivar G2333 which offers broad resistance for anthracnose has been widely used as a source of resistance in breeding for anthracnose resistance. The cultivar is pyramided with three genes namely CO-4, CO-5 and CO-7 and of these three genes, the CO-4 gene has been found to offer the broadest resistance. The main aim of this work was to identify and validate easily assayable PCR based co-dominant molecular markers for selection of the CO-4 gene in segregating populations derived from crosses of G2333 with RWR 1946 and RWR 2075, two commercial Andean cultivars highly susceptible to anthracnose. Marker sequences for the study were obtained by blasting the sequence of the COK-4 gene in the Phaseolus gene database. Primer sequence pairs that were not provided from the Phaseolus gene database were designed by the use of Primer3 software. PCR conditions were optimized and the PCR products were run on 6% HPAGE gel. Results of the polymorphism test indicated that out of 18 identified markers, only two markers namely BM588 and BM211 behaved co-dominantly. Phenotypic evaluation for reaction to anthracnose disease was done by inoculating 21days old seedlings of three parents, F1 and F2 populations with race 7 of Colletotrichum lindemuthianum in the humid chamber. DNA testing of the BM588 marker onto the F2 segregating population of the crosses RWR 1946 x G 2333 and RWR 2075 x G2333 further revealed that the marker BM588 co-segregated with disease resistance with co-dominance of two alleles of 200bp and 400bp, fitting the expected segregation ratio of 1:2:1. The BM588 marker was significantly associated with disease resistance and gave promising results for marker assisted selection of the CO-4 gene in the breeding lines. Activities to validate the BM211 marker are also underway.

Keywords: codominant, Colletotrichum lindemuthianum, MAS, Phaseolus vulgaris

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3198 Association of Brain-Derived Neurotrophic Factor (BDNF) Gene with Obesity and Metabolic Traits in Malaysian Adults

Authors: Yamunah Devi Apalasamy, Sanjay Rampal, Tin Tin Su, Foong Ming Moy, Hazreen Abdul Majid, Awang Bulgiba, Zahurin Mohamed

Abstract:

Obesity is a growing global health issue. Obesity results from a combination of environmental and genetics factors. Brain-derived neurotrophic factor (BDNF), a gene encodes the BDNF protein and the BDNF gene have been linked to regulation of body weight and appetite. Genome-wide association studies have identified the BDNF variants to be related to obesity among Caucasians, East Asians, and Filipinos. However, the role of BDNF in other ethnic groups remains inconclusive. This case control study aims to investigate the associations of BDNF gene polymorphisms with obesity and metabolic parameters in Malaysian Malays. BDNF rs4074134, BDNF rs10501087 and BDNF rs6265 were genotyped using Sequenom MassARRAY. Anthropometric, body fat, fasting lipids and glucose levels were measured. A total of 663 subjects (194 obese and 469 non-obese) were included in this study. There were no significant associations association between BDNF SNPs and obesity. The allelic and genotype frequencies of the BDNF SNPs were similar in the obese and non-obese groups. After adjustment for age and sex, the BDNF variants were not associated with obesity, body fat, fasting lipids and glucose levels. Haplotypes at the BDNF gene region, were not significantly associated with obesity. The BDNF rs4074134 was in strong LD with BDNF rs10501087 (D'=0.98) and BDNF rs6265 (D'=0.87). The BDNF rs10501087 was also in strong LD with BDNF rs6265 (D'=0.91). Our findings suggest that the BDNF variants and the haplotypes of BDNF gene were not associated with obesity and metabolic traits in this study population. Further research is needed to explore other BDNF variants with a larger sample size with gene-environment interactions in multi ethnic Malaysian population.

Keywords: genomics of obesity, SNP, BMI, haplotypes

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3197 The Alarming Caesarean-Section Delivery Rate in Addis Ababa, Ethiopia

Authors: Yibeltal T. Bayou, Yohana S. Mashalla, Gloria Thupayagale-Tshweneagae

Abstract:

Background: According to the World Health Organization, caesarean section delivery rates of more than 10-15% caesarean section deliveries in any specific geographic region in the world are not justifiable. The aim of the study was to describe the level and analyse determinants of caesarean section delivery in Addis Ababa. Methods: Data was collected in Addis Ababa using a structured questionnaire administered to 901 women aged 15-49 years through a stratified two-stage cluster sampling technique. Binary logistic regression model was employed to identify predictors of caesarean section delivery. Results: Among the 835 women who delivered their last birth at healthcare facilities, 19.2% of them gave birth by caesarean section. About 9.0% of the caesarean section births were due to mother’s request or service provider’s influence without any medical indication. The caesarean section delivery rate was much higher than the recommended rate particularly among the non-slum residents (27.2%); clients of private healthcare facilities (41.1%); currently married women (20.6%); women with secondary (22.2%) and tertiary (33.6%) level of education; and women belonging to the highest wealth quintile household (28.2%). The majority (65.8%) of the caesarean section clients were not informed about the consequences of caesarean section delivery by service providers. The logistic regression model shows that older age (30-49), secondary and above education, non-slum residence, high-risk pregnancy and receiving adequate antenatal care were significantly positively associated with caesarean section delivery. Conclusion: Despite the unreserved effort towards achieving MDG 5 through safe skilled delivery assistance among others, the high caesarean section rate beyond the recommend limit, and the finding that caesarean sections done without medical indications were also alarming. The government and city administration should take appropriate measures before the problems become setbacks in healthcare provision. Further investigations should focus on the effect of caesarean section delivery on maternal and child health outcomes in the study area.

Keywords: Addis Ababa, caesarean section, mode of delivery, slum residence

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3196 New Isolate of Cucumber Mosaic Virus Infecting Banana

Authors: Abdelsabour G. A. Khaled, Ahmed W. A. Abdalla And Sabry Y. M. Mahmoud

Abstract:

Banana plants showing typical mosaic and yellow stripes on leaves as symptoms were collected from Assiut Governorate in Egypt. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of symptoms, transmission, serology, transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Coat protein (CP) gene was amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. In this study the CMV was transmitted into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA). Analysis of the 120 deduced amino acid sequence of the coat protein gene revealed that the EG-A strain of CMV shared from 97.50 to 98.33% with those strains belonging to subgroup IA. The cluster analysis grouped the Egyptian isolate with strains Fny and Ri8 belonging sub-group IA. It appears that there occurs a high incidence of CMV infecting banana belonging to IA subgroup in most parts of Egypt.

Keywords: banana, CMV, transmission, CP gene, RT-PCR

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3195 A Gene Selection Algorithm for Microarray Cancer Classification Using an Improved Particle Swarm Optimization

Authors: Arfan Ali Nagra, Tariq Shahzad, Meshal Alharbi, Khalid Masood Khan, Muhammad Mugees Asif, Taher M. Ghazal, Khmaies Ouahada

Abstract:

Gene selection is an essential step for the classification of microarray cancer data. Gene expression cancer data (DNA microarray) facilitates computing the robust and concurrent expression of various genes. Particle swarm optimization (PSO) requires simple operators and less number of parameters for tuning the model in gene selection. The selection of a prognostic gene with small redundancy is a great challenge for the researcher as there are a few complications in PSO based selection method. In this research, a new variant of PSO (Self-inertia weight adaptive PSO) has been proposed. In the proposed algorithm, SIW-APSO-ELM is explored to achieve gene selection prediction accuracies. This new algorithm balances the exploration capabilities of the improved inertia weight adaptive particle swarm optimization and the exploitation. The self-inertia weight adaptive particle swarm optimization (SIW-APSO) is used to search the solution. The SIW-APSO is updated with an evolutionary process in such a way that each particle iteratively improves its velocities and positions. The extreme learning machine (ELM) has been designed for the selection procedure. The proposed method has been to identify a number of genes in the cancer dataset. The classification algorithm contains ELM, K- centroid nearest neighbor (KCNN), and support vector machine (SVM) to attain high forecast accuracy as compared to the start-of-the-art methods on microarray cancer datasets that show the effectiveness of the proposed method.

Keywords: microarray cancer, improved PSO, ELM, SVM, evolutionary algorithms

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3194 Unravelling the Relationship Between Maternal and Fetal ACE2 Gene Polymorphism and Preeclampsia Risk

Authors: Sonia Tamanna, Akramul Hassan, Mohammad Shakil Mahmood, Farzana Ansari, Gowhar Rashid, Mir Fahim Faisal, M. Zakir Hossain Howlader

Abstract:

Background: Preeclampsia (PE), a pregnancy-specific hypertensive disorder, significantly impacts maternal and fetal health. It is particularly prevalent in underdeveloped countries and is linked to preterm delivery and fetal growth. The renin-angiotensin system (RAS) plays a crucial role in ensuring a successful pregnancy outcome, with Angiotensin-Converting Enzyme 2 (ACE2) being a key component. ACE2 converts ANG II to Ang-(1-7), offering protection against ANG II-induced stress and inflammation while regulating blood pressure and osmotic balance during pregnancy. The reduced maternal plasma angiotensin-converting enzyme 2 (ACE2) seen in preeclampsia might contribute to its pathogenesis. However, there has been a dearth of comprehensive research into the association between ACE2 gene polymorphism and preeclampsia. In the South Asian population, hypertension is strongly linked to two SNPs: rs2285666 and rs879922. This genotype was therefore considered, and the possible association of maternal and fetal ACE2 gene polymorphism with preeclampsia within the Bangladeshi population was evaluated. Method: DNA was extracted from peripheral white blood cells (WBCs) using the organic method, and SNP genotyping was done via PCR-RFLP. Odds ratios (OR) with 95% confidence intervals (95% CI) were calculated using logistic regression to determine relative risk. Result: A comprehensive case-control study was conducted on 51 PE patients and their infants, along with 56 control subjects and their infants. Maternal single nuvleotide polymorphisms (SNP) (rs2285666) analysis revealed a strong association between the TT genotype and preeclampsia, with a four-fold increased risk in mothers (P=0.024, OR=4.00, 95% CI=1.36-11.37) compared to their ancestral genotype CC. However, the CT genotype (rs2285666) showed no significant difference (P=0.46, OR=1.54, 95% CI=0.57-4.14). Notably, no significant correlation was found in infants, regardless of their gender. For rs879922, no significant association was observed in both mothers and infants. This pioneering study suggests that mothers carrying the ACE2 gene variant rs2285666 (TT allele) may be at higher risk for preeclampsia, potentially influencing hypertension characteristics, whereas rs879922 does not appear to be associated with developing preeclampsia. Conclusion: This study sheds light on the role of ACE2 gene polymorphism, particularly the rs2285666 TT allele, in maternal susceptibility to preeclampsia. However, rs879922 does not appear to be linked to the risk of PE. This research contributes to our understanding of the genetic underpinnings of preeclampsia, offering insights into potential avenues for prevention and management.

Keywords: ACE2, PCR-RFLP, preeclampsia, single nuvleotide polymorphisms (SNPs)

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3193 Fruiting Body Specific Sc4 Hydrophobin Gene Plays a Role in Schizophyllum Commune Hyphal Attachment to Structured Glass Surfaces

Authors: Evans Iyamu

Abstract:

Genes encoding hydrophobins play distinct roles at different stages of the life cycle of fungi, and they foster hyphal attachment to surfaces. The hydrophobin Sc4 is known to provide a hydrophobic membrane lining of the gas channels within Schizophyllum commune fruiting bodies. Here, we cultivated non-fruiting, monokaryotic S. commune 12-43 on glass surfaces that could be verified by micrography. Differential gene expression profiling of nine hydrophobin genes and the hydrophobin-like sc15 gene by quantitative PCR showed significant up-regulation of sc4 when S. commune was attached to glass surfaces, also confirmed with RNA-Seq data analysis. Another silicate, namely quartz sand, was investigated, and induction of sc4 was seen as well. The up-regulation of the hydrophobin gene sc4 may indicate involvement in S. commune hyphal attachment to glass as well as quartz surfaces. We propose that the covering of hyphae by Sc4 allows for direct interaction with the hydrophobic surfaces of silicates and that differential functions of specific hydrophobin genes depend on the surface interface involved. This study could help with the clarification of the biological functions of hydrophobins in natural surroundings, including hydrophobic surface attachment. Therefore, the analysis of growth on glass serves as a basis for understanding S. commune interaction with glass surfaces while providing the possibility to visualize the interaction microscopically.

Keywords: hydrophobin, structured glass surfaces, differential gene expression, quartz sand

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3192 Quantitative Evaluation of Endogenous Reference Genes for ddPCR under Salt Stress Using a Moderate Halophile

Authors: Qinghua Xing, Noha M. Mesbah, Haisheng Wang, Jun Li, Baisuo Zhao

Abstract:

Droplet digital PCR (ddPCR) is being increasingly adopted for gene detection and quantification because of its higher sensitivity and specificity. According to previous observations and our lab data, it is essential to use endogenous reference genes (RGs) when investigating gene expression at the mRNA level under salt stress. This study aimed to select and validate suitable RGs for gene expression under salt stress using ddPCR. Six candidate RGs were selected based on the tandem mass tag (TMT)-labeled quantitative proteomics of Alkalicoccus halolimnae at four salinities. The expression stability of these candidate genes was evaluated using statistical algorithms (geNorm, NormFinder, BestKeeper and RefFinder). There was a small fluctuation in cycle threshold (Ct) value and copy number of the pdp gene. Its expression stability was ranked in the vanguard of all algorithms, and was the most suitable RG for quantification of expression by both qPCR and ddPCR of A. halolimnae under salt stress. Single RG pdp and RG combinations were used to normalize the expression of ectA, ectB, ectC, and ectD under four salinities. The present study constitutes the first systematic analysis of endogenous RG selection for halophiles responding to salt stress. This work provides a valuable theory and an approach reference of internal control identification for ddPCR-based stress response models.

Keywords: endogenous reference gene, salt stress, ddPCR, RT-qPCR, Alkalicoccus halolimnae

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3191 Iontophoretic Drug Transport of Some Anti-Diabetic Agents

Authors: Ashish Jain, Satish Nayak

Abstract:

Transdermal iontophoretic drug delivery system is viable drug delivery platform technology and has a strong market worldwide. Transdermal drug delivery system is particularly desirable for therapeutic agents that need prolonged administration at controlled plasma level. This makes appropriateness to antihypertensive and anti-diabetic agents for their transdermal development. Controlled zero order absorption, easily termination of drug delivery and easy to administration also support for popularity of transdermal delivery. In this current research iontophoretic delivery of various anti diabetic agents like glipizide, glibenclamide and glimepiride were carried out. The experiments were carried out at different drug concentrations and different current densities using cathodal iontophoresis. Diffusion cell for iontophoretic permeation study was modified according to Glikfield Design. Pig skin was used for in vitro permeation study and for the in-vivo study New Zealand rabbits were used. At all concentration level iontophoresis showed enhanced permeation rate compared to passive controls. Iontophoretic transports of selected drugs were found to be increased with the current densities. Results showed that target permeation rate for selected drugs could be achieved with the aid of iontophoresis by increasing the area in an appreciable range.

Keywords: transdermal, iontophoresis, pig skin, rabbits, glipizide, glibeclamide

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3190 Gene Expression Profile Reveals Breast Cancer Proliferation and Metastasis

Authors: Nandhana Vivek, Bhaskar Gogoi, Ayyavu Mahesh

Abstract:

Breast cancer metastasis plays a key role in cancer progression and fatality. The present study examines the potential causes of metastasis in breast cancer by investigating the novel interactions between genes and their pathways. The gene expression profile of GSE99394, GSE1246464, and GSE103865 was downloaded from the GEO data repository to analyze the differentially expressed genes (DEGs). Protein-protein interactions, target factor interactions, pathways and gene relationships, and functional enrichment networks were investigated. The proliferation pathway was shown to be highly expressed in breast cancer progression and metastasis in all three datasets. Gene Ontology analysis revealed 11 DEGs as gene targets to control breast cancer metastasis: LYN, DLGAP5, CXCR4, CDC6, NANOG, IFI30, TXP2, AGTR1, MKI67, and FTH1. Upon studying the function, genomic and proteomic data, and pathway involvement of the target genes, DLGAP5 proved to be a promising candidate due to it being highly differentially expressed in all datasets. The study takes a unique perspective on the avenues through which DLGAP5 promotes metastasis. The current investigation helps pave the way in understanding the role DLGAP5 plays in metastasis, which leads to an increased incidence of death among breast cancer patients.

Keywords: genomics, metastasis, microarray, cancer

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3189 Mutational Analysis of DNase I Gene in Diabetic Patients

Authors: Hateem Zafar Kayani, Nageen Hussain

Abstract:

The main aim is to analyze the mutations of DNASE I gene in diabetic patients. A total of 120 diabetes patients and 120 controls were sampled. The total number of male diabetic patients included in the study was 79 (66%) while female patients were 41 (34%) in number. Exon 8 of the DNASE I gene was amplified by using thermo cycler. The possible band of interest was located at 165 base pairs. Two samples showed similar missense mutations at 127th position of exon 8 which replaced amino acid Arginine (Arg) to Glutamine (Gln). All controls showed no mutations. The association of diabetes with different levels of blood pressure and body mass index (BMI) were found to be significant.

Keywords: deoxyribonuclease I, polymerase chain reaction, insulin-dependent diabetes mellitus, non-insulin dependent diabetes mellitus

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3188 Men's Decision Making: The Determinant of Home Delivery among Women in Khyber Pakhtunkhwa Pakistan

Authors: Hussain Ali, Ahmad Ali, Syed Rashid Ali

Abstract:

The maternal mortality is one of the basic health issues faced by rural women in Pakistan. There are various structural and socio-cultural determinants which confine women to domestic sphere. Such mobility restriction compels women for home delivery which causes high maternal mortality and morbidity. However, it is hard to find out the research findings and well-organized literature that explain the cultural factors act as determinant to home delivery among Pakhtun women. The overall objective of this research is to study men’s decision making within the household in Pakhtun society as determinant of home delivery among Pakhtun women in Khyber Pakhtunkhwa province of Pakistan. In the present study, researchers used the quantitative research design in which the data are collected through household survey technique from (n=503) ever-married women having reproductive age (15-49 years) by using interview schedule. The data are analyzed through SPSS, and binary logistic regression was applied to draw the association between home as a place of delivery and men’s decision making in the Pakhtun society. The results show that majority (76%) of the husbands are key decision makers about the home delivery due to their superior position within household. Similarly, majority (88%) Pakhtun women prefer to stay in home for their delivery due to their dependency on husband’s decision. The researcher concludes that men are key decision makers in Pakhtun society and their decisions affect women maternal health care. Similarly, the women are in subordinate position, and their limited decision making in the domestic sphere are greatly responsible for home delivery which causing high maternal mortality rate in the study area. In order to achieve Sustainable Development Goal No. 3, the study recommends empowering women in the decision making about accessing and utilizing maternal health care services and given financial autonomy to them.

Keywords: home delivery, men’s decision, Pakhtun women, subordinate position

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3187 Prognostic Implication of Nras Gene Mutations in Egyptian Adult Acute Myeloid Leukemia

Authors: Doaa M. Elghannam, Nashwa Khayrat Abousamra, Doaa A. Shahin, Enas F. Goda, Hanan Azzam, Emad Azmy, Manal Salah El-Din

Abstract:

Background: The pathogenesis of acute myeloid leukemia (AML) involves the cooperation of mutations promoting proliferation/survival and those impairing differentiation. Point mutations of the NRAS gene are the most frequent somatic mutations causing aberrant signal-transduction in acute myeloid leukemia (AML). Aim: The present work was conducted to study the frequency and prognostic significance of NRAS gene mutations (NRASmut) in de novo Egyptian adult AML. Material and methods: Bone marrow specimens from 150 patients with de novo acute myeloid leukemia and controls were analyzed by genomic PCR-SSCP at codons 12, 13 (exon 1), and 61 (exon 2) for NRAS mutations. Results: NRAS gene mutations was found in 19/150 (12.7%) AML cases, represented more frequently in the FAB subtype M4eo (P = 0.028), and at codon 12, 13 (14of 19; 73.7%). Patients with NRASmut had a significant lower peripheral marrow blasts (P = 0.004, P=0.03) and non significant improved clinical outcome than patients without the mutation. Complete remission rate was (63.2% vs 56.5%; p=0.46), resistant disease (15.8% vs 23.6%; p=0.51), three years overall survival (44% vs 42%; P = 0.85) and disease free survival (42.1% vs 38.9%, P = 0.74). Multivariate analysis showed that age was the strongest unfavorable factor for overall survival (relative risk [RR], 1.9; P = .002), followed by cytogenetics (P = .004). FAB types, NRAS mutation, and leukocytosis were less important. Conclusions: NRAS gene mutation frequency and spectrum differ between biologically distinct subtypes of AML but do not significantly influence prognosis and clinical outcome.

Keywords: NRAS Gene, egyptian adult, acute myeloid leukemia, cytogenetics

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