Search results for: essential genes
4770 Isolation and Molecular Characterization of Lytic Bacteriophage against Carbapenem Resistant Klebsiella pneumoniae
Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla
Abstract:
Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,
Procedia PDF Downloads 1864769 Microorganism and Laurus nobilis from Mascara - Algeria
Authors: Karima Oldyerou, B. Meddah, A. Tirtouil
Abstract:
Laurusnobilis is an aromatic plant, common in Algeria and widely used by local people as a source of spice and for medicinal purposes. The essential oil of this plant is the subject of this work in a physicochemical and microbiological study. The extraction of the essential oil was carried by steam distillation and the highest yield (1.5%) was determined in May. The organoleptic and physico-chemical characters are consistent with those obtained in the literature with some differences that can be attributed to certain factors. Evaluation of antibacterial activity showed a sensitivity of Salmonella spp. with an MIC of 2,5 mg.ml-1, and other bacteria of the intestinal flora of Wistar rats: E. coli and Lactobacillus sp. have a high potential for resistance with MICs respectively equal to 10 and 20 mg.ml-1.Keywords: laurus nobilis, essential oil, physicochemical character, MIC, intestinal flora, antibacterial activity
Procedia PDF Downloads 3274768 Disturbed Cellular Iron Metabolism Genes in Neurodevelopmental Disorders is Different from Neurodegenerative Disorders
Authors: O. H. Gebril, N. A. Meguid
Abstract:
Background: Iron had been a focus of interest recently as a main exaggerating factor for oxidative stresses in the central nervous system and a link to various neurological disorders is suspected. Many studies with various techniques showed evidence of disturbed iron-related proteins in the cell in human and animal models of neurodegenerative disorders. Also, linkage to significant pathological changes had been evidenced e.g. apoptosis and cell signaling. On the other hand, the role of iron in neurodevelopmental disorders is still unclear. With increasing prevalence of autism worldwide, some changes in iron parameters and its stores were documented in many studies. This study includes Haemochromatosis HFE gene polymorphisms (p.H63D and p.C282Y) and ferroportin gene (SLC40A1) Q248H polymorphism in autism and control children. Materials and Methods: Whole genome DNA was extracted; p.H63D and p.C282Y genotyping was studied using specific sequence amplification followed by restriction enzyme digestion on a sample of autism patients (25 cases) and twenty controls. Results: The p.H63D is seen more than the C282Y among both autism and control samples, with no significant association of p.H63D or p.C282Y polymorphism and autism was revealed. Also, no association with Q248H polymorphism was evidenced. Conclusion: The study results do not prove the role of cellular iron genes polymorphisms as risk factors for neurodevelopmental disorders, and in turn highlights the specificity of cellular iron related pathways in neurodegeneration. These results demand further gene expression studies to elucidate the main pathophysiological pathways that are disturbed in autism and other neurodevelopmental disorders.Keywords: iron, neurodevelopmental, oxidative stress, haemohromatosis, ferroportin, genes
Procedia PDF Downloads 3574767 Environmental Efficacy on Heracleum persicum Essential Oils
Authors: Rahele Hasani, Iraj Mehregan, Kambiz Larijani, Taher Nejadsattari, Romain Scalone
Abstract:
Essential oils of Heracleum persicum (Apiaceae) have been widely used from many years ago, but the difference of its properties among different populations have not been identified up to now. Hydrodistilation Clevenger type was used to obtaining the fruit essential oils of four populations of H. persicum from different localities in Iran, then they were characterized by GC-FID and GC-MS analyses. Some ecological factors were also measured. The oils of four populations were compared to determine the similarities and differences and the relationships between these factors and ecological factors. Based on the result, 18-32 different components were identified in four populations, while the percentage of the main components was higher in population with lower number of components. According to the statistical analyses of chemical components and ecological factors, it can be concluded that some ecological factors such as altitude, less humidity, high difference between day and night temperature and salty soil would lead to lower number of components in essential oil, whereas they consist the higher percentage.Keywords: Chemotaxonomy, Persian hogweed, Ecological factors, Apiaceae.
Procedia PDF Downloads 4284766 Genomic Surveillance of Bacillus Anthracis in South Africa Revealed a Unique Genetic Cluster of B- Clade Strains
Authors: Kgaugelo Lekota, Ayesha Hassim, Henriette Van Heerden
Abstract:
Bacillus anthracis is the causative agent of anthrax that is composed of three genetic groups, namely A, B, and C. Clade-A is distributed world-wide, while sub-clades B has been identified in Kruger National Park (KNP), South Africa. KNP is one of the endemic anthrax regions in South Africa with distinctive genetic diversity. Genomic surveillance of KNP B. anthracis strains was employed on the historical culture collection isolates (n=67) dated from the 1990’s to 2015 using a whole genome sequencing approach. Whole genome single nucleotide polymorphism (SNPs) and pan-genomics analysis were used to define the B. anthracis genetic population structure. This study showed that KNP has heterologous B. anthracis strains grouping in the A-clade with more prominent ABr.005/006 (Ancient A) SNP lineage. The 2012 and 2015 anthrax isolates are dispersed amongst minor sub-clades that prevail in non-stabilized genetic evolution strains. This was augmented with non-parsimony informative SNPs of the B. anthracis strains across minor sub-clades of the Ancient A clade. Pan-genomics of B. anthracis showed a clear distinction between A and B-clade genomes with 11 374 predicted clusters of protein coding genes. Unique accessory genes of B-clade genomes that included biosynthetic cell wall genes and multidrug resistant of Fosfomycin. South Africa consists of diverse B. anthracis strains with unique defined SNPs. The sequenced B. anthracis strains in this study will serve as a means to further trace the dissemination of B. anthracis outbreaks globally and especially in South Africa.Keywords: bacillus anthracis, whole genome single nucleotide polymorphisms, pangenomics, kruger national park
Procedia PDF Downloads 1374765 Evaluation of Gene Expression after in Vitro Differentiation of Human Bone Marrow-Derived Stem Cells to Insulin-Producing Cells
Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim
Abstract:
Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique
Procedia PDF Downloads 2074764 Isolation and Identification of Salmonella spp and Salmonella enteritidis, from Distributed Chicken Samples in the Tehran Province using Culture and PCR Techniques
Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri
Abstract:
Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.Keywords: culture, PCR, salmonella spp, salmonella enteritidis
Procedia PDF Downloads 654763 Difference in Virulence Factor Genes Between Transient and Persistent Streptococcus Uberis Intramammary Infection in Dairy Cattle
Authors: Anyaphat Srithanasuwan, Noppason Pangprasit, Montira Intanon, Phongsakorn Chuammitri, Witaya Suriyasathaporn, Ynte H. Schukken
Abstract:
Streptococcus uberis is one of the most common mastitis-causing pathogens, with a wide range of intramammary infection (IMI) durations and pathogenicity. This study aimed to compare shared or unique virulence factor gene clusters distinguishing persistent and transient strains of S. uberis. A total of 139 S. uberis strains were isolated from three small-holder dairy herds with a high prevalence of S. uberis mastitis. The duration of IMI was used to categorize bacteria into two groups: transient and persistent strains with an IMI duration of less than 1 month and longer than 2 months, respectively. Six representative S. uberis strains, three from each group (transience and persistence) were selected for analysis. All transient strains exhibited multi-locus sequence types (MLST), indicating a highly diverse population of transient S. uberis. In contrast, MLST of persistent strains was available in an online database (pubMLST). Identification of virulence genes was performed using whole-genome sequencing (WGS) data. Differences in genomic size and number of virulent genes were found. For example, the BCA gene or alpha-c protein and the gene associated with capsule formation (hasAB), found in persistent strains, are important for attachment and invasion, as well as the evasion of the antimicrobial mechanisms and survival persistence, respectively. These findings suggest a genetic-level difference between the two strain types. Consequently, a comprehensive study of 139 S. uberis isolates will be conducted to perform an in-depth genetic assessment through WGS analysis on an Illumina platform.Keywords: Streptococcus Uberis, mastitis, whole genome sequence, intramammary infection, persistent S. Uberis, transient s. Uberis
Procedia PDF Downloads 524762 Identification of Conserved Domains and Motifs for GRF Gene Family
Authors: Jafar Ahmadi, Nafiseh Noormohammadi, Sedegeh Fabriki Ourang
Abstract:
GRF, Growth regulating factor, genes encode a novel class of plant-specific transcription factors. The GRF proteins play a role in the regulation of cell numbers in young and growing tissues and may act as transcription activations in growth and development of plants. Identification of GRF genes and their expression are important in plants to performance of the growth and development of various organs. In this study, to better understanding the structural and functional differences of GRFs family, 45 GRF proteins sequences in A. thaliana, Z. mays, O. sativa, B. napus, B. rapa, H. vulgare, and S. bicolor, have been collected and analyzed through bioinformatics data mining. As a result, in secondary structure of GRFs, the number of alpha helices was more than beta sheets and in all of them QLQ domains were completely in the biggest alpha helix. In all GRFs, QLQ, and WRC domains were completely protected except in AtGRF9. These proteins have no trans-membrane domain and due to have nuclear localization signals act in nuclear and they are component of unstable proteins in the test tube.Keywords: domain, gene family, GRF, motif
Procedia PDF Downloads 4534761 Selection of Potential Starter Using Their Transcription Level
Authors: Elif Coskun Daggecen, Seyma Dokucu, Yekta Gezginc, Ismail Akyol
Abstract:
Fermented dairy food quality is mainly determined by the sensory perception and influenced by many factors. Today, starter cultures for fermented foods are being developed to have a constant quality in these foods. Streptococcus thermophilus is one of the main species of most a starter cultures of yogurt fermentation. This species produces lactate by lactose fermentation from pyruvate. On the other hand, a small amount of pyruvate can alternatively be converted to various typical yoghurt flavor compounds such as diacetyl, acetoin, acetaldehyde, or acetic acid, for which the activity of three genes are shown to be especially important; ldh, nox and als. Up to date, commercially produced yoghurts have not yet met the desired aromatic properties that Turkish consumers find in traditional homemade yoghurts. Therefore, it is important to select starters carrying favorable metabolic characteristics from natural isolates. In this study, 30 strains of Str. Thermophilus were isolated from traditional Turkish yoghurts obtained from different regions of the country. In these strains, transcriptional levels of ldh, nox and als genes were determined via a newly developed qPCR protocol, which is a more reliable and precision method for analyzing the quantitative and qualitative expression of specific genes in different experimental conditions or in different organisms compared to conventional analytical methods. Additionally, the metabolite production potentials of the isolates were measured. Of all the strains examined, 60% were found to carry the metabolite production potential and the gene activity which appeared to be suitable to be used as a starter culture. Probable starter cultures were determined according to real-time PCR results.Keywords: gene expression, RT-PCR, starter culture, Streptococcus thermophilus
Procedia PDF Downloads 1824760 Biofilm Formation Due to the Proteome Changes Of Enterococcus Faecium in Response to Sub-Mic of Gentamicin
Authors: Amin Abbasi, Mahdi Asghari Ozma
Abstract:
Background and Objective:Enterococcus faecium is a normal flora of the human gastrointestinal tract that causes infection in the host body under conditions such as biofilm formation, in which the use of antibiotics causes changes in these pathogenic mechanisms. In this study, we aimed to evaluate comprehensively the changes in E.faecium when exposed to sub-MIC of the gentamicin,especiallythe biofilm formation rate. Materials and Methods: For this study, the keywords "Enterococcus faecium ", "Biofilm", and "Gentamicin" in the databases PubMed, Google Scholar, Sid, and MagIran between 2015 and 2021 were searched, and 14 articles were chosen, studied, and analyzed. Results: Gentamicin significantly had increased biofilm formation in most of the isolates in the studies. Increased expression of the genes (efaA and esp) and proteins involved in biofilm formation and decreased expression of the genes (gelE and cylA) involved in spreading and proteins involved in metabolism and cell division in E.faecium were the most significant cause of the biofilm formation, which were increased in sub-MIC gentamicin-treated situation. Conclusion: Inadequate use of gentamicin intensify biofilm formation of E.faecium, which can make the treatment of infections caused by this bacterium difficult.Keywords: biofilm, enterococcus faecium, gentamicin, proteome
Procedia PDF Downloads 1054759 Transcriptome Sequencing of the Spleens Reveals Genes Involved in Antiviral Response in Chickens Infected with Castv
Authors: Sajewicz-Krukowska Joanna, Domańska-Blicharz Katarzyna, Tarasiuk Karolina, Marzec-Kotarska Barbara
Abstract:
Astroviral infections pose a significant problem in the poultry industry, leading to multiple adverse effects such as decreased egg production, breeding disorders, poor weight gain, and even increased mortality. Commonly observed chicken astrovirus (CAstV) was recently reported to be responsible for "white chicks syndrome" associated with increased embryo/chick mortality. The CAstV-mediated pathogenesis in chicken occurs due to complex interactions between the infectious pathogen and the immune system. Many aspects of CAstV-chicken interactions remain unclear, and there is no information available regarding gene expression changes in the chicken's spleen in response to CAstV infection. We aimed to investigate the molecular background triggered by CAstV infection. Ten 21-day-old SPF White Leghorn chickens were divided into two groups of 5 birds each. One group was inoculated with CAstV, and the other was used as the negative control. On 4th dpi, spleen samples were collected and immediately frozen at -70°C for RNA isolation. We analysed transcriptional profiles of the chickens' spleens at the 4th day following infection using RNA-seq to establish differentially expressed genes (DEGs). The RNA-seq findings were verified by quantitative real-time PCR (qRT-PCR). A total of 31959 transcripts were identified in response to CAstV infection. Eventually 45 DEGs (p-value<0.05; Log2Foldchange>1)were recognized in the spleen after CAstV infection (26 upregulated DEGs and 19 downregulated DEGs). qRT-PCR performed on 4 genes (IFIT5, OASL, RASD1, DDX60) confirmed RNAseq results. Top differentially expressed genes belonged to novel putative IFN-induced CAstV restriction factors. Most of the DEGs were associated with RIG-I–like signalling pathway or, more generally, with an innate antiviral response(upregulated: BLEC3, CMPK2, IFIT5, OASL, DDX60, IFI6, and downregulated: SPIK5, SELENOP, HSPA2, TMEM158, RASD1, YWHAB). The study provided a global analysis of host transcriptional changes that occur during CAstV infection in vivo and proved the cell cycle in the spleen and immune signalling in chickens were predominantly affected upon CAstV infection.Keywords: chicken astrovirus, CastV, RNA-seq, transcriptome, spleen
Procedia PDF Downloads 1464758 Chemical Composition and Antimicrobial Activity of the Essential Oil of Thymus lanceolatus Desf. an Endemic Thyme from Algeria
Authors: Ahmed Nouasri, Tahar Dob, Toumi Mohamed, Dahmane Dahmane, Soumioa Krimat, Lynda Lamari, Chabane Chelghom
Abstract:
The aim of this study is to investigate the chemical composition for the first time, and antimicrobial activities of essential oil (EO) of Thymus lanceolatus Desf., an endemic thyme from Tiaret province of Algeria. The chemical composition of hydrodistilled essential oil from flowering aerial parts has been analyzed by GC and GC/MS techniques, the antimicrobial activity was realised by agar disc diffusion method and MIC was determined in solid medium by direct contact. Essential oil of T. lanceolataus has been yielded of 2.336 (w/w) based on dry weight, the analyses cited above, led to the identification of 29 components, which accounted for 97.34% of the total oil. Oxygenated monoterpenes was the main fraction (88.31%) dominated by thymol (80.2%) as major component of this oil, followed by carvacrol (6.25%). The oil was found effective against all tested strains especially fungus, except Pseudomonas aeruginosa were low activity observed, in addition Gram (+) bacteria found to be more sensitive to the EO than Gram (-) bacteria. This activity was ranging from12±2.65mm to 60.00±0.00mm Ø, with the lowest MIC value of under 0.06mg/ml to 12.53mg/ml. This results provided the evidence that the studied plant might indeed be potential sources of natural antimicrobial agentsKeywords: Thymus lanceolatus Desf., essential oil, chemical composition, antimicrobial activities
Procedia PDF Downloads 4674757 Significance of Molecular Autophagic Pathway in Gaucher Disease Pathology
Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik
Abstract:
Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts
Procedia PDF Downloads 3214756 Chemical Composition, Antioxidant and Antibacterial Activities of Essential Oil from the Leaves of Thymus vulgaris L.
Authors: Tsige Reda
Abstract:
Essential oil of Thymus vulgaris was extracted by means of hydro-distillation. This study was done to investigate the chemical composition, antibacterial and antioxidant activities. The chemical composition of the essential oils was determined using gas chromatography coupled to mass spectroscopy (GC-MS). Using disc diffusion assay the antibacterial activity was assessed on one Gram-positive bacteria and one Gram-negative bacteria. The percentage oil yield of the essential oil was found to be 0.97 ± 0.08% (w/w) with yellow color. The physicochemical constants of the oil were also noted. The phytochemical screening of the plant extract revealed the presence of tannins, saponins, phenol, flavonoids, terpenoids, steroids and alkaloids. A total of 18 chemical constituents were identified by Gas Chromatography-Mass Spectroscopy analysis representing 100% of the total essential oil of Thymus vulgaris, with thymol (31.977%), o-cymene (29.992%), and carvacrol (14.541%). Previous studies have revealed that the thymol, o-cymen and carvacrol components of Thymus vulgaris are responsible for their biological activities. Thymus vulgaris have been used traditionally to treat a wide variety of infections. Based on the extensive use and lack of scientific evidence, a study was embarked upon to determine its bioactivity. The essential oil of Thymus vulgaris leaves exhibited higher activity towards the Gram-positive bacteria (Staphylococcus aurous) than the Gram-negative bacteria (Escherichia coli) and also has good antioxidant activity, and can be used medicinal and therapeutic applications. This activity may be due to the high amount of thymol, o-cymen and carvacrol.Keywords: hydro-distillation, Thymus vulgaris, essential oil composition, phytochemical screening, physicochemical constants, antioxidant activity, antibacterial activity
Procedia PDF Downloads 4334755 Effects of Essential Oils on the Intestinal Microflora of Termite (Heterotermes indicola)
Authors: Ayesha Aihetasham, Najma Arshad, Sobia Khan
Abstract:
Damage causes by subterranean termites are of major concern today. Termites majorly treated with pesticides resulted in several problems related to health and environment. For this reason, plant-derived natural products specifically essential oils have been evaluated in order to control termites. The aim of the present study was to investigate the antitermitic potential of six essential oils on Heterotermes indicola subterranean termite. No-choice bioassay was used to assess the termiticidal action of essential oils. Further, gut from each set of treated termite group was extracted and analyzed for reduction in number of protozoa and bacteria by protozoal count method using haemocytometer and viable bacterial plate count (dilution method) respectively. In no-choice bioassay it was found that Foeniculum vulgare oil causes high degree of mortality 90 % average mortality at 10 mg oil concentration (10mg/0.42g weight of filter paper). Least mortality appeared to be due to Citrus sinensis oil (43.33 % average mortality at 10 mg/0.42g). The highest activity verified to be of Foeniculum vulgare followed by Eruca sativa, Trigonella foenum-graecum, Peganum harmala, Syzygium cumini and Citrus sinensis. The essential oil which caused maximum reduction in number of protozoa was P. harmala followed by T. foenum-graecum and E. sativa. In case of bacterial count E. sativa oil indicated maximum decrease in bacterial number (6.4×10⁹ CFU/ml). It is concluded that F. vulgare, E. sativa and P. harmala essential oils are highly effective against H. indicola termite and its gut microflora.Keywords: bacterial count, essential oils, Heterotermes indicola, protozoal count
Procedia PDF Downloads 2394754 Microbiological Study of Two Spontaneous Plants of Algerian Sahara Septentrional: Cotula cinerea and Chamomilla recutita
Authors: Mehani Mouna, Boukhari Nadjet, Ladjal Segni
Abstract:
The aim of our study is to determine the antimicrobial effect of essential oils of two plants Cotula cinerea and Chamomilla recutita on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Humans use plants for thousands of years to treat various ailments, in many developing countries; much of the population relies on traditional doctors and their collections of medicinal plants to cure them. The test adopted is based on the diffusion method on solid medium (Antibiogram), this method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plants Cotula cinerea and Chamomilla recutita have a different effect on the resistance of germs.Keywords: antibiogram, Chamomilla recutita, Cotula cinerea, essential oil, microorganism
Procedia PDF Downloads 3074753 Comparison between Two Groups of Pathogenic Bacteria under Different Essential Oil Extract of Ocimum basilicum L.
Authors: A. M. Daneshian Moghaddam, J. Shayegh, J. Dolghari Sharaf
Abstract:
This study was conducted to assessment the antibacterial activities of different part of basil essential oil on the standard gram-negative bacteria include Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and gram-positive ones including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogen. The basil essential oil was provided from two part of plant (leaf and herb) at the two different developmental stage. The antibacterial properties of basil essential oil was studied Also agar disk diffusion, minimal inhibition concentration (MIC) and minimum bactericidal concentration (MBC) were detected. The results of agar disk diffusion tests showed the inhibition zones as follow: Listeria monocytogen 17.11-17.42 mm, St. aureus 29.20-30.56 mm, B. cereus 14.73-16.06 mm, E. coli 21.60-23.58 mm, Salmonella typhi 21.63-24.80 mm and for P. aeruginosa the maximum inhibition zones were seen on leaf essential oil. From the herb part of basil almost similar results were obtained: Listeria monocytogen 17.02-17.67 mm, St. aureus 29.60-30.41 mm, B. cereus 10.66-16.11 mm, E. coli 17.48-23.54 mm, Salmonella typhi 21.58-21.64 mm and for P. aeruginosa the maximum inhibition zones were seen. The MICs for gram-positive bacteria were as: B. cereus ranging 36-18 μg/mL, S. aureus 18 μg/mL, Listeria monocytogen 18-36 μg/mL and for gram-negative bacteria of E. coli, Salmonella typhi and P. aeruginosa were 18-9 μg/mL.Keywords: basil (Ocimum basilicum) essential oil, gram-positive and gram negative bacteria, antibacterial activity, MIC, MBC
Procedia PDF Downloads 4384752 Relationship Between tcdA and tcdB Genes of Clostridium difficile with Duration of Diarrhea in Elderly Patients
Authors: Ni Luh Putu Harta Wedari
Abstract:
Background: Clostridium difficile has two main virulence factors, namely TcdA and TcdB. TcdA encoded by the tcdA gene acts as an enterotoxin, pro-inflammatory and fluid accumulation, while TcdB encoded by the tcdB gene is cytotoxic, causes disruption of the actin cytoskeleton, and causes disruption of tight junctions in colon cells. This study aims to explore the relationship between the tcdA and tcdB genes and the duration of diarrhea in elderly patients. Method: This research was an observational analytic with a prospective cross-sectional with samples of elderly diarrhea patients who met the inclusion criteria in Denpasar City health service facilities from 1 December 2022 until 30 June 2023, and then their feces were analyzed using the real-time PCR method. Results: In this study, 40 elderly diarrhea patients met the inclusion criteria and in accordance with the minimum sample size, 28 (70%) men and 12 (30%) women. 5 patients (12.5%) had a history of azithromycin, 4 (10%) levofloxacin, 17 (42.5%) ciprofloxacin, 8 (20%) metronidazole, 1 (2.5%) cefoperazone, 5 (12, 5%) doxycycline. Comorbids, namely 13 (32.5%) type II diabetes mellitus, 4 (10%) chronic kidney disease, 10 (25%) malignancies, 7 (17.5%) urinary tract infections, 3 (7.5%) %) immunocompromised, 2 (5%) cardiac heart failure, and 1 (2.5%) acute on chronic kidney disease. The overall diarrhea duration average was 5 days. 8 samples (20%) were positive for 16s rRNA, and there was no significant difference in diarrhea duration with negative samples (p=0.166). The relationship between the tcdA gene and the duration of diarrhea could not be performed because all samples were negative. Likewise, relationship analysis between the coexistence of tcdA and tcdB could not be performed. There was no significant difference between tcdB positive 3 (7.5%) and negative with diarrhea duration (p=0.739). Conclusion: There is no significant relationship between the presence of the 16s rRNA and tcdB C. difficile genes with the duration of diarrhea in elderly patients.Keywords: clostridium, difficile, diarrhea, elderly, tcdA, tcdB
Procedia PDF Downloads 704751 The Effects of Soil Parameters on Efficiency of Essential Oil from Zingiber zerumbet (L.) Smith in Thailand
Authors: Worakrit Worananthakij, Kamonchanok Doungtadum, Nattagan Mingkwan, Supatsorn Chupong
Abstract:
Natural products from herb have been used in different aspects of life as a result of their various biological activities. Generally, plant growth and production of secondary compounds largely depend on environmental conditions. To better understand this correlation, study on biological activity and soil parameter is necessary. This research aims to study the soil parameters which affect the efficiency of the antioxidant activity of essential oils extracted from the Zingiber zerumbet in three areas of Thailand, including Min Buri district, Bangkok province; Muang district, Chiang Mai province and Kaeng Sanam Nang district, Nakhon Ratchasima province. The soil samples in each area were collected and analyzed in the laboratory. The essential oil of Z. zerumbet in each province was extracted and tested for antioxidant activity by hydrodistillation method and DPPH (2,2-diphenyl-1-picrylhydrazyl radical) assay, respectively. The results showed that, the soil parameters such as pH, nitrogen, potassium and phosphorus elements and exchange of cations of soil specimen from Nakhon Ratchasima province were the highest (P<0.05) (6.10 ±0.03, 0.15 ± 0.04 percent of total nitrogen, 16.67 ± 0.46 mg/L, 3.35 ± 0.65 mg/kg and 12.87 ± 0.11 cmol/kg, respectively). In addition, IC50 (Inhibition Concentrtion of antioxidant at 50%) of Z. zerumbet essential oil collected from Nakhon Ratchasima showed the highest value (P<0.05) (1,400 µg/mL). In conclusion, the soil parameters are once important factor for the efficiency of essential oils extract from Z. zerumbet.Keywords: antioxidant, essential oil, herb, soil parameter, Zingiber zerumbet
Procedia PDF Downloads 2374750 Integrated Risk Management in The Supply Chain of Essential Medicines in Zambia
Authors: Mario M. J. Musonda
Abstract:
Access to health care is a human right, which includes having timely access to affordable and quality essential medicines at the right place and in sufficient quantity. However, inefficient public sector supply chain management contributes to constant shortages of essential medicines at health facilities. Literature review involved a desktop study of published research studies and reports on risk management, supply chain management of essential medicines and their integration to increase the efficiency of the latter. The research was conducted on a sample population of offices under Ministry of Health Headquarters, Lusaka Provincial and District Offices, selected health facilities in Lusaka, Medical Stores Limited, Zambia Medicines Regulatory Authority and Cooperating Partners. Individuals involved in study were selected judgmentally by their functions under selection and quantification, regulation, procurement, storage, distribution, quality assurance, and dispensing of essential medicines. Structured interviews and discussions were held with selected experts and self-administered questionnaires were distributed. Collected and analysed data of 35 returned and usable questionnaires from the 50 distributed. The highest prioritised risks were; inadequate and inconsistent fund disbursements, weak information management systems, weak quality management systems and insufficient resources (HR and infrastructure) among others. The results for this research can be used to increase the efficiency of the public sector supply chain of essential medicines and other pharmaceuticals. The results of the study showed that there is need to implement effective risk management systems by participating institutions and organisations to increase the efficiency of the entire supply chain in order to avoid and/or reduce shortages of essential medicines at health facilities.Keywords: essential medicine, risk assessment, risk management, supply chain, supply chain risk management
Procedia PDF Downloads 4384749 Development of Essential Oil-Loaded Gelatin Hydrogels for Use as Antibacterial Wound Dressing
Authors: Piyachat Chuysinuan, Nitirat Chimnoi, Arthit Makarasen, Nanthawan Reuk-Ngam, Pitt Supaphol, Supanna Techasakul
Abstract:
In this work, biomaterial wound dressings was developed based on gelatin containing herbal substances (essential oil), a substance from the plant Eupatorium adenophorum Spreng (Crofton weed) that used as traditional wound healers. Gelatin hydrogel was prepared from a 10 wt-% gelatin solution. The oil in water (o/w) emulsion Eupatorium adenophorum of essential oil were prepared and used Pluronic F68 as a surfactant. The 10, 20, and 30 % v/v emulsion were mixed with gelatin solution and cast into film. These hydrogels were tested for their gel fraction, swelling and weight loss behavior. With an increase in the emulsion concentration the emulsion-loaded in hydrogels, the gel fraction were decreased due to the crosslink density, while the swelling and weight loss behavior were increased with an increasing in the emulsion content. The potential to use the emulsion-containing gelatin hydrogels as wound dressing was assessed on investigation the release characteristics of the as-loaded hydrogels. The E. adenophorum essential oil was first identified the chemical composition by using GC-MS analysis. The principal components of the oil were p-cymene (16.23%), bornyl acetate (11.84%), and amorpha-4, 7(11)-diene (10.51%). The hydrogel wound dressing containing essential oil was then characterized for their antibacterial activity against Gram-positive and Gram-negative in order to elucidate their potential for use as antibacterial wound dressings by using agar disk diffusion methods. The result showed that E. adenophorum essential oil and the emulsion-loaded gelatin hydrogel inhibited the growth of the test pathogens, Staphylococcus aureus and Staphylococcus epidermidis and increased with increasing the initial amount of essential oil in the hydrogels which confirmed their application as antibacterial wound dressings. Furthermore, the potential use of these wound dressings was further assessed in terms of the indirect cytotoxicity, in vitro attachment and proliferation of dermal human fibroblasts cultured in the hydrogel wound dressings.Keywords: hydrogel, antibacterial wound dressing, Eupatorium adenophorum essential oil, gelatin
Procedia PDF Downloads 3484748 Impact of Ocean Acidification on Gene Expression Dynamics during Development of the Sea Urchin Species Heliocidaris erythrogramma
Authors: Hannah R. Devens, Phillip L. Davidson, Dione Deaker, Kathryn E. Smith, Gregory A. Wray, Maria Byrne
Abstract:
Marine invertebrate species with calcifying larvae are especially vulnerable to ocean acidification (OA) caused by rising atmospheric CO₂ levels. Acidic conditions can delay development, suppress metabolism, and decrease the availability of carbonate ions in the ocean environment for skeletogenesis. These stresses often result in increased larval mortality, which may lead to significant ecological consequences including alterations to the larval settlement, population distribution, and genetic connectivity. Importantly, many of these physiological and developmental effects are caused by genetic and molecular level changes. Although many studies have examined the effect of near-future oceanic pH levels on gene expression in marine invertebrates, little is known about the impact of OA on gene expression in a developmental context. Here, we performed mRNA-sequencing to investigate the impact of environmental acidity on gene expression across three developmental stages in the sea urchin Heliocidaris erythrogramma. We collected RNA from gastrula, early larva, and 1-day post-metamorphic juvenile sea urchins cultured at present-day and predicted future oceanic pH levels (pH 8.1 and 7.7, respectively). We assembled an annotated reference transcriptome encompassing development from egg to ten days post-metamorphosis by combining these data with datasets from two previous developmental transcriptomic studies of H. erythrogramma. Differential gene expression and time course analyses between pH conditions revealed significant alterations to developmental transcription that are potentially associated with pH stress. Consistent with previous investigations, genes involved in biomineralization and ion transport were significantly upregulated under acidic conditions. Differences in gene expression between the two pH conditions became more pronounced post-metamorphosis, suggesting a development-dependent effect of OA on gene expression. Furthermore, many differences in gene expression later in development appeared to be a result of broad downregulation at pH 7.7: of 539 genes differentially expressed at the juvenile stage, 519 of these were lower in the acidic condition. Time course comparisons between pH 8.1 and 7.7 samples also demonstrated over 500 genes were more lowly expressed in pH 7.7 samples throughout development. Of the genes exhibiting stage-dependent expression level changes, over 15% of these diverged from the expected temporal pattern of expression in the acidic condition. Through these analyses, we identify novel candidate genes involved in development, metabolism, and transcriptional regulation that are possibly affected by pH stress. Our results demonstrate that pH stress significantly alters gene expression dynamics throughout development. A large number of genes differentially expressed between pH conditions in juveniles relative to earlier stages may be attributed to the effects of acidity on transcriptional regulation, as a greater proportion of mRNA at this later stage has been nascent transcribed rather than maternally loaded. Also, the overall downregulation of many genes in the acidic condition suggests that OA-induced developmental delay manifests as suppressed mRNA expression, possibly from lower transcription rates or increased mRNA degradation in the acidic environment. Further studies will be necessary to determine in greater detail the extent of OA effects on early developing marine invertebrates.Keywords: development, gene expression, ocean acidification, RNA-sequencing, sea urchins
Procedia PDF Downloads 1624747 A Comparison of Antibiotic Resistant Enterobacteriaceae: Diabetic versus Non-Diabetic Infections
Authors: Zainab Dashti, Leila Vali
Abstract:
Background: The Middle East, in particular Kuwait, contains one of the highest rates of patients with Diabetes in the world. Generally, infections resistant to antibiotics among the diabetic population has been shown to be on the rise. This is the first study in Kuwait to compare the antibiotic resistance profiles and genotypic differences between the resistant isolates of Enterobacteriaceae obtained from diabetic and non-diabetic patients. Material/Methods: In total, 65 isolates were collected from diabetic patients consisting of 34 E. coli, 15 K. pneumoniae and 16 other Enterobacteriaceae species (including Salmonella spp. Serratia spp and Proteus spp.). In our control group, a total of 49 isolates consisting of 37 E. coli, 7 K. pneumoniae and 5 other species (including Salmonella spp. Serratia spp and Proteus spp.) were included. Isolates were identified at the species level and antibiotic resistance profiles, including Colistin, were determined using initially the Vitek system followed by double dilution MIC and E-test assays. Multi drug resistance (MDR) was defined as isolates resistant to a minimum of three antibiotics from three different classes. PCR was performed to detect ESBL genes (blaCTX-M, blaTEM & blaSHV), flouroquinolone resistance genes (qnrA, qnrB, qnrS & aac(6’)-lb-cr) and carbapenem resistance genes (blaOXA, blaVIM, blaGIM, blaKPC, blaIMP, & blaNDM) in both groups. Pulse field gel electrophoresis (PFGE) was performed to compare clonal relatedness of both E. coli and K.pneumonaie isolates. Results: Colistin resistance was determined in three isolates with MICs of 32-128 mg/L. A significant difference in resistance to ampicillin (Diabetes 93.8% vs control 72.5%, P value <0.002), augmentin (80% vs 52.5%, p value < 0.003), cefuroxime (69.2% vs 45%, p value < 0.0014), ceftazadime (73.8% vs 42.5%, p value <0.001) and ciprofloxacin (67.6% vs 40%, p value < 0.005) were determined. Also, a significant difference in MDR rates between the two groups (Diabetes 76.9%, control 57.5%, p value <0.036 were found. All antibiotic resistance genes showed a higher prevalence among the diabetic group, except for blaCTX-M, which was higher among the control group. PFGE showed a high rate of diversity between each group of isolates. Conclusions: Our results suggested an alarming rate of antibiotic resistance, in particular Colistin resistance (1.8%) among K. pneumoniea isolated from diabetic patients in Kuwait. MDR among Enterobacteriaceae infections also seems to be a worrying issue among the diabetics of Kuwait. More efforts are required to limit the issue of antibiotic resistance in Kuwait, especially among patients with diabetes mellitus.Keywords: antibiotic resistance, diabetes, enterobacreriacae, multi antibiotic resistance
Procedia PDF Downloads 3604746 Elucidation of Mechanism of Action of Antidepressant-Like Effect of Valeriana wallichii Maaliol Chemotype in Mice
Authors: Sangeeta Pilkhwal Sah, C. S. Mathela, Kanwaljit Chopra
Abstract:
Valeriana wallichii DC, an ayurvedic traditional medicine, popularly named as Indian valerian exist as three chemotypes. GC-MS analysis of V. wallichii essential oil in present study showed maaliol as the major constituent followed by the presence of β-gurjunene, acoradiene, guaiol and α-santalene. The results thus confirmed it to be a maaliol chemotype. Further, the antidepressant-like effect of root essential oil (10, 20 and 40 mg/kg p.o.) was investigated in both acute and chronic treatment study using forced swim test in mice. Single administration of different doses produced an inverted U shaped curve and significantly inhibited the immobility period (39.7% and 58%) at doses 10 and 40 mg/kg respectively. Standard drug imipramine significantly decreased immobility period (59.8%). None of the doses altered locomotor activity except a significant decrease of 44.9% was observed with 40 mg/kg (p < 0.05). Similarly, daily administration of essential oil for 14 days produced a dose dependent effect with significantly reduced immobility period (70.9%) at 40 mg/kg dose only whereas imipramine produced 86% decrease (p < 0.05). The neurotransmitter levels in mouse brain were estimated on day 14 after the behavioral study. Significant increase in the level of norepinephrine (10%) and dopamine (23%) (p < 0.05) was found at 40 mg/kg dose, while no change was observed at 10 and 20 mg/kg doses. The antidepressant-like effect of essential oil (40 mg/kg) was prevented by pretreatment of mice with L-arginine (750 mg/kg i.p.) and sildenafil (5 mg/kg i.p). On the contrary, pretreatment of mice with L-NAME (10 mg/kg i.p.) or methylene blue (10 mg/kg i.p.) potentiated the antidepressant action of essential oil (20 mg/kg). The findings thus demonstrated that nitric oxide pathway is involved in mediating antidepressant like effect of essential oil from this chemotype.Keywords: Valeriana wallichii DC chemotype, essential oil, forced swim test, nitric oxide modulators, neurotransmitters
Procedia PDF Downloads 2934745 Seed Dressing and Foliar Spray of Green Bean (Phaseolus vulgaris L.) with Essential Oils and Disinfectants for Suppressing Root Rot and Wilt Incidence under Field Conditions
Authors: N. S. El-Mougy, M. M. Abdel-Kader, H. M. Abouelnasr
Abstract:
The efficacy of two essential oils applied as a bean seed dressing followed by seedlings foliar spray with four commercial disinfectants against root rot and wilt incidence was evaluated under field conditions. The essential oils, grape seed and peppermint oils and the disinfectants, Gold, Revarest, Klenva, Malva were applied. Chitosan and the fungicide Topsin-M were used as comparison treatment. Essential oils individually or combined with disinfectants were applied as a bean seed dressing. Furthermore, emerged bean plants were sprayed with the same treatments. Under laboratory conditions, growth inhibition effect was observed for the isolated, tested fungi R. solani and F. oxysporum when exposed to essential oils individually or combined with disinfectants. A high inhibitor effect was recorded for peppermint followed by grape seed oils. Concentrations of 1% and 2% of chitosan as well as Topsin M at 400 ppm showed complete reduction (100%) in the two fungal growths. Under field conditions, the obtained results showed that the applied treatments of chitosan had a superior effect on root rot and wilt disease incidence compared with other tested treatments. It was found that seed coating treatment provides good protection of emerged green bean seeds against the root pathogens attack compared with the fungicide and control treatments. Also, the application of seed dressing with essential oils accompanied by seedling spray demonstrated similar results. It was observed that essential oils had an enhancing effect against disease incidence when combined with disinfectants compared with their application. The obvious yield increase was significantly higher in all applied treatments than in fungicide and control.Keywords: bean, disinfectants, essential oils, root rot, wilt
Procedia PDF Downloads 1544744 Differential Expression Profile Analysis of DNA Repair Genes in Mycobacterium Leprae by qPCR
Authors: Mukul Sharma, Madhusmita Das, Sundeep Chaitanya Vedithi
Abstract:
Leprosy is a chronic human disease caused by Mycobacterium leprae, that cannot be cultured in vitro. Though treatable with multidrug therapy (MDT), recently, bacteria reported resistance to multiple antibiotics. Targeting DNA replication and repair pathways can serve as the foundation of developing new anti-leprosy drugs. Due to the absence of an axenic culture medium for the propagation of M. leprae, studying cellular processes, especially those belonging to DNA repair pathways, is challenging. Genomic understanding of M. Leprae harbors several protein-coding genes with no previously assigned function known as 'hypothetical proteins'. Here, we report identification and expression of known and hypothetical DNA repair genes from a human skin biopsy and mouse footpads that are involved in base excision repair, direct reversal repair, and SOS response. Initially, a bioinformatics approach was employed based on sequence similarity, identification of known protein domains to screen the hypothetical proteins in the genome of M. leprae, that are potentially related to DNA repair mechanisms. Before testing on clinical samples, pure stocks of bacterial reference DNA of M. leprae (NHDP63 strain) was used to construct standard graphs to validate and identify lower detection limit in the qPCR experiments. Primers were designed to amplify the respective transcripts, and PCR products of the predicted size were obtained. Later, excisional skin biopsies of newly diagnosed untreated, treated, and drug resistance leprosy cases from SIHR & LC hospital, Vellore, India were taken for the extraction of RNA. To determine the presence of the predicted transcripts, cDNA was generated from M. leprae mRNA isolated from clinically confirmed leprosy skin biopsy specimen across all the study groups. Melting curve analysis was performed to determine the integrity of the amplification and to rule out primer‑dimer formation. The Ct values obtained from qPCR were fitted to standard curve to determine transcript copy number. Same procedure was applied for M. leprae extracted after processing a footpad of nude mice of drug sensitive and drug resistant strains. 16S rRNA was used as positive control. Of all the 16 genes involved in BER, DR, and SOS, differential expression pattern of the genes was observed in terms of Ct values when compared to human samples; this was because of the different host and its immune response. However, no drastic variation in gene expression levels was observed in human samples except the nth gene. The higher expression of nth gene could be because of the mutations that may be associated with sequence diversity and drug resistance which suggests an important role in the repair mechanism and remains to be explored. In both human and mouse samples, SOS system – lexA and RecA, and BER genes AlkB and Ogt were expressing efficiently to deal with possible DNA damage. Together, the results of the present study suggest that DNA repair genes are constitutively expressed and may provide a reference for molecular diagnosis, therapeutic target selection, determination of treatment and prognostic judgment in M. leprae pathogenesis.Keywords: DNA repair, human biopsy, hypothetical proteins, mouse footpads, Mycobacterium leprae, qPCR
Procedia PDF Downloads 994743 Fatty Acid and Amino Acid Composition in Mene maculata in The Sea of Maluku
Authors: Semuel Unwakoly, Reinner Puppela, Maresthy Rumalean, Healthy Kainama
Abstract:
Fish is a kind of food that contains many nutritions, one of those is the long chain of unsaturated fatty acids as omega-3 and omega-6 fatty acids and essential amino acid in enough amount for the necessity of our body. Like pelagic fish that found in the sea of Maluku. This research was done to identify fatty acids and amino acids composition in Moonfish (M. maculata) using transesterification reaction steps and Gas Chromatograph-Mass Spectrophotometer (GC-MS) and High-Performance Liquid Chromatography (HPLC). The result showed that fatty acids composition in Moonfish (M. maculata) contained tridecanoic acid (2.84%); palmitoleic acid (2.65%); palmitic acid (35.24%); oleic acid (6.2%); stearic acid (14.20%); and 5,8,11,14-eicosatetraenoic acid (1.29%) and 12 amino acids composition that consist of 7 essential amino acids, were leucine, isoleucine, valine, phenylalanine, methionine, lysine, and histidine, and also 5 non-essential amino acid, were tyrosine, glycine, alanine, glutamic acid, and arginine.Thus, these fishes can be used by the people to complete the necessity of essential fatty acid and amino acid.Keywords: Moonfish (M. maculata), fatty acid, amino acid, GC-MS, HPLC
Procedia PDF Downloads 2424742 Expression of ACSS2 Genes in Peripheral Blood Mononuclear Cells of Patients with Alzheimer’s Disease
Authors: Ali Bayram, Burak Uz, Remzi Yiğiter
Abstract:
The impairment of lipid metabolism in the central nervous system has been suggested as a critical factor of Alzheimer’s disease (AD) pathogenesis. Homo sapiens acyl-coenyme A synthetase short-chain family member 2 (ACSS2) gene encodes the enzyme acetyl-Coenzyme A synthetase (AMP forming; AceCS) providing acetyl-coenzyme A (Ac-CoA) for various physiological processes, such as cholesterol and fatty acid synthesis, as well as the citric acid cycle. We investigated ACSS2, transcript variant 1 (ACSS2*1), mRNA levels in the peripheral blood mononuclear cells (PBMC) of patients with AD and compared them with the controls. The study group comprised 50 patients with the diagnosis of AD who have applied to Gaziantep University Faculty of Medicine, and Department of Neurology. 49 healthy individuals without any neurodegenerative disease are included as controls. ACSS2 mRNA expression in PBMC of AD/control patients was 0.495 (95% confidence interval: 0.410-0.598), p= .000000001902). Further studies are needed to better clarify this association.Keywords: Alzheimer’s disease, ACSS2 Genes, mRNA expression, RT-PCR
Procedia PDF Downloads 3834741 Stability of a Biofilm Reactor Able to Degrade a Mixture of the Organochlorine Herbicides Atrazine, Simazine, Diuron and 2,4-Dichlorophenoxyacetic Acid to Changes in the Composition of the Supply Medium
Authors: I. Nava-Arenas, N. Ruiz-Ordaz, C. J. Galindez-Mayer, M. L. Luna-Guido, S. L. Ruiz-López, A. Cabrera-Orozco, D. Nava-Arenas
Abstract:
Among the most important herbicides, the organochlorine compounds are of considerable interest due to their recalcitrance to the chemical, biological, and photolytic degradation, their persistence in the environment, their mobility, and their bioacummulation. The most widely used herbicides in North America are primarily 2,4-dichlorophenoxyacetic acid (2,4-D), the triazines (atrazine and simazine), and to a lesser extent diuron. The contamination of soils and water bodies frequently occurs by mixtures of these xenobiotics. For this reason, in this work, the operational stability to changes in the composition of the medium supplied to an aerobic biofilm reactor was studied. The reactor was packed with fragments of volcanic rock that retained a complex microbial film, able to degrade a mixture of organochlorine herbicides atrazine, simazine, diuron and 2,4-D, and whose members have microbial genes encoding the main catabolic enzymes atzABCD, tfdACD and puhB. To acclimate the attached microbial community, the biofilm reactor was fed continuously with a mineral minimal medium containing the herbicides (in mg•L-1): diuron, 20.4; atrazine, 14.2, simazine, 11.4, and 2,4-D, 59.7, as carbon and nitrogen sources. Throughout the bioprocess, removal efficiencies of 92-100% for herbicides, 78-90% for COD, 92-96% for TOC and 61-83% for dehalogenation were reached. In the microbial community, the genes encoding catabolic enzymes of different herbicides tfdACD, puhB and, occasionally, the genes atzA and atzC were detected. After the acclimatization, the triazine herbicides were eliminated from the mixture formulation. Volumetric loading rates of the mixture 2,4-D and diuron were continuously supplied to the reactor (1.9-21.5 mg herbicides •L-1 •h-1). Along the bioprocess, the removal efficiencies obtained were 86-100% for the mixture of herbicides, 63-94% for for COD, 90-100% for COT, and dehalogenation values of 63-100%. It was also observed that the genes encoding the enzymes in the catabolism of both herbicides, tfdACD and puhB, were consistently detected; and, occasionally, the atzA and atzC. Subsequently, the triazine herbicide atrazine and simazine were restored to the medium supply. Different volumetric charges of this mixture were continuously fed to the reactor (2.9 to 12.6 mg herbicides •L-1 •h-1). During this new treatment process, removal efficiencies of 65-95% for the mixture of herbicides, 63-92% for COD, 66-89% for TOC and 73-94% of dehalogenation were observed. In this last case, the genes tfdACD, puhB and atzABC encoding for the enzymes involved in the catabolism of the distinct herbicides were consistently detected. The atzD gene, encoding the cyanuric hydrolase enzyme, could not be detected, though it was determined that there was partial degradation of cyanuric acid. In general, the community in the biofilm reactor showed some catabolic stability, adapting to changes in loading rates and composition of the mixture of herbicides, and preserving their ability to degrade the four herbicides tested; although, there was a significant delay in the response time to recover to degradation of the herbicides.Keywords: biodegradation, biofilm reactor, microbial community, organochlorine herbicides
Procedia PDF Downloads 433