Search results for: gene diversity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3077

Search results for: gene diversity

1997 Conservation Status of a Lowland Tropical Forest in South-West, Nigeria

Authors: Lucky Dartsa Wakawa, Friday Nwabueze Ogana, Temitope Elizabeth Adeniyi

Abstract:

Timely and reliable information on the status of a forest is essential for assessing the extent of regeneration and degradation. However, when such information is lacking effective forest management practices becomes impossible. Therefore, this study assessed the tree species composition, richness, diversity, structure of Oluwa forest reserve with the view of ascertaining it conservation status. A systematic line transect was used in the laying of eight (8) temporary sample plots (TSPs) of size 50m x 50m. Trees with Dbh ≥ 10cm in the selected plots were enumerated, identified and measured. The results indicate that 535 individual trees were enumerated cutting across 26 families and 58 species. The family Sterculiaceae recorded the highest number of species (10) and occurrence (112) representing 17.2% and 20.93% respectively. Celtis zenkeri is the species with the highest number of occurrence of tree per hectare and importance value index (IVI) of 59 and 53.81 respectively. The reserve has the Margalef's index of species richness, Shannon-Weiner diversity Index (H') and Pielou's Species Evenness Index (EH) of 9.07, 3.43 and 0.84 respectively. The forest has a mean Dbh (cm), mean height (m), total basal area/ha (m2) and total volume/ha (m3) of 24.7, 16.9, 36.63 and 602.09 respectively. The important tropical tree species identified includes Diospyros crassiflora Milicia excels, Mansonia altisima, Triplochiton scleroxylon. Despite the level of exploitation in the forest, the forest seems to be resilience. Given the right attention, it could regenerate and replenish to save some of the original species composition of the reserve.

Keywords: forest conservation, forest structure, Lowland tropical forest, South-west Nigeria

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1996 Collagen Deposition in Lung Parenchyma Driven by Depletion of LYVE-1+ Macrophages Protects Emphysema and Loss of Airway Function

Authors: Yinebeb Mezgebu Dagnachew, Hwee Ying Lim, Liao Wupeng, Sheau Yng Lim, Lim Sheng Jie Natalie, Veronique Angeli

Abstract:

Collagen is essential for maintaining lung structure and function, and its remodeling has been associated with respiratory diseases, including chronic obstructive pulmonary disease (COPD). However, the cellular mechanisms driving collagen remodeling and the functional implications of this process in the pathophysiology of pulmonary diseases remain poorly understood. Using a mouse model of Lyve-1 expressing macrophage depletion, we found that the absence of this subpopulation of tissue-resident macrophage led to the preferential deposition of type I collagen fibers around the alveoli and bronchi in the steady state. Further analysis by polarized light microscopy revealed that the collagen fibers accumulating in the lungs depleted of Lyve-1+ macrophages were thicker and crosslinked. A decrease in MMP-9 gene expression and proteolytic activity, together with an increase in Col1a1, Timp-3 and Lox gene expression, accompanied the collagen alterations. Next, we investigated the effect of the collagen remodeling on the pathophysiology of COPD and airway function in mouse lacking Lyve-1+ macrophage exposed chronically to cigarette smoke (CS), a well-established animal model of COPD. We showed that the deposition of collagen protected mouse against the destruction of alveoli (emphysema) and bronchi thickening after CS exposure and prevented loss of airway function. Thus, we demonstrate that interstitial Lyve-1+ macrophages regulate the composition, amount, and architecture of the collagen network in the lungs and that such collagen remodeling functionally impacts the development of COPD. This study further supports the potential of targeting collagen as a promising approach to treating respiratory diseases.

Keywords: lung, extracellular matrix, chronic obstructive pulmonary disease, matrix metalloproteinases, collagen

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1995 Effect of Graded Level of Nano Selenium Supplementation on the Performance of Broiler Chicken

Authors: Raj Kishore Swain, Kamdev Sethy, Sumanta Kumar Mishra

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Selenium is an essential trace element for the chicken with a variety of biological functions like growth, fertility, immune system, hormone metabolism, and antioxidant defense systems. Selenium deficiency in chicken causes exudative diathesis, pancreatic dystrophy and nutritional muscle dystrophy of the gizzard, heart and skeletal muscle. Additionally, insufficient immunity, lowering of production ability, decreased feathering of chickens and increased embryo mortality may occur due to selenium deficiency. Nano elemental selenium, which is bright red, highly stable, soluble and of nano meter size in the redox state of zero, has high bioavailability and low toxicity due to the greater surface area, high surface activity, high catalytic efficiency and strong adsorbing ability. To assess the effect of dietary nano-Se on performance and expression of gene in Vencobb broiler birds in comparison to its inorganic form (sodium selenite), four hundred fifty day-old Vencobb broiler chicks were randomly distributed into 9 dietary treatment groups with two replicates with 25 chicks per replicate. The dietary treatments were: T1 (Control group): Basal diet; T2: Basal diet with 0.3 ppm of inorganic Se; T3: Basal diet with 0.01875 ppm of nano-Se; T4: Basal diet with 0.0375 ppm of nano-Se; T5: Basal diet with 0.075 ppm of nano-Se, T6: Basal diet with 0.15 ppm of nano-Se, T7: Basal diet with 0.3 ppm of nano-Se, T8: Basal diet with 0.60 ppm of nano-Se, T9: Basal diet with 1.20 ppm of nano-Se. Nano selenium was synthesized by mixing sodium selenite with reduced glutathione and bovine serum albumin. The experiment was carried out in two phases: starter phase (0-3 wks), finisher phase (4-5 wk) in deep litter system. The body weight at the 5th week was best observed in T4. The best feed conversion ratio at the end of 5th week was observed in T4. Erythrocytic catalase, glutathione peroxidase and superoxide dismutase activity were significantly (P < 0.05) higher in all the nano selenium treated groups at 5th week. The antibody titers (log2) against Ranikhet diseases vaccine immunization of 5th-week broiler birds were significantly higher (P < 0.05) in the treatments T4 to T7. The selenium levels in liver, breast, kidney, brain, and gizzard were significantly (P < 0.05) increased with increasing dietary nano-Se indicating higher bioavailability of nano-Se compared to inorganic Se. The real time polymer chain reaction analysis showed an increase in the expression of antioxidative gene in T4 and T7 group. Therefore, it is concluded that supplementation of nano-selenium at 0.0375 ppm over and above the basal level can improve the body weight, antioxidant enzyme activity, Se bioavailability and expression of the antioxidative gene in broiler birds.

Keywords: chicken, growth, immunity, nano selenium

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1994 Black-Brown and Yellow-Brown-Red Skin Pigmentation Elements are Shared in Common: Using Art and Science for Multicultural Education

Authors: Mary Kay Bacallao

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New research on the human genome has revealed secrets to the variation in skin pigmentation found in all human populations. Application of this research to multicultural education has a profound effect on students from all backgrounds. This paper identifies the four locations in the human genome that code for variation in skin pigmentation worldwide. The research makes this new knowledge accessible to students of all ages as they participate in an art project that brings these scientific multicultural concepts to life. Students participate in the application of breakthrough scientific principles through hands-on art activities where they simulate the work of the DNA coding to create their own skin tone using the colors expressed to varying degrees in every people group. As students create their own artwork handprint from the pallet of colors, they realize that each color on the pallet is essential to creating every tone of skin. This research project serves to bring people together and appreciate the variety and diversity in skin tones. As students explore the variations, they create pigmentation with the use of the eumelanins, which are the black-brown sources of pigmentation, and the pheomelanins, which are the yellow-reddish-brown sources of pigmentation. The research project dispels myths about skin tones that have divided people in the past. As a group project, this research leads to greater appreciation and understanding of the diverse family groups.

Keywords: diversity, multicultural, skin pigmentation, eumelanins, pheomelanins, handprint, artwork, science, genome, human

Procedia PDF Downloads 56
1993 An Intelligent Scheme Switching for MIMO Systems Using Fuzzy Logic Technique

Authors: Robert O. Abolade, Olumide O. Ajayi, Zacheaus K. Adeyemo, Solomon A. Adeniran

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Link adaptation is an important strategy for achieving robust wireless multimedia communications based on quality of service (QoS) demand. Scheme switching in multiple-input multiple-output (MIMO) systems is an aspect of link adaptation, and it involves selecting among different MIMO transmission schemes or modes so as to adapt to the varying radio channel conditions for the purpose of achieving QoS delivery. However, finding the most appropriate switching method in MIMO links is still a challenge as existing methods are either computationally complex or not always accurate. This paper presents an intelligent switching method for the MIMO system consisting of two schemes - transmit diversity (TD) and spatial multiplexing (SM) - using fuzzy logic technique. In this method, two channel quality indicators (CQI) namely average received signal-to-noise ratio (RSNR) and received signal strength indicator (RSSI) are measured and are passed as inputs to the fuzzy logic system which then gives a decision – an inference. The switching decision of the fuzzy logic system is fed back to the transmitter to switch between the TD and SM schemes. Simulation results show that the proposed fuzzy logic – based switching technique outperforms conventional static switching technique in terms of bit error rate and spectral efficiency.

Keywords: channel quality indicator, fuzzy logic, link adaptation, MIMO, spatial multiplexing, transmit diversity

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1992 RNA-seq Analysis of Liver from NASH-HCC Model Mouse Treated with Streptozotocin-High Fat Diet

Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

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Non-alcoholic steatohepatitis (NASH) is a chronic liver disease, often associated with type II diabetes, which sometimes progresses to more serious conditions such as liver fibrosis and hepatocellular carcinoma (HCC). NASH has become an important health problem worldwide, buttherapeutic agents for NASH have not yet been approved, and animal models with high clinical correlation are required. TheSTAM™ mouse shows the same pathological progression as human NASH patients and has been widely used for both drug efficacy and basic research, such as lipid profiling and gut microbiota research. In this study, we analyzed the RNA-seq data of STAM™mice at each pathological stage (steatosis, steatohepatitis, liver fibrosis, and HCC) and examined the clinical correlation at the genetic level. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin solution 2 days after birth and feeding with high fat dietafter 4 weeks of age. The mice were sacrificed and livers collected at 6, 8, 10, 12, 16, and 20 weeks of age. For liver samples, the left lateral lobe was snap frozen in liquid nitrogen and stored at -80˚C for RNA-seq analysis. Total RNA of the cells was isolated using RNeasy mini kit. The gene expression of the canonical pathways in NASH progression from steatosis to hepatocellular carcinoma were analyzed, such as immune system process, oxidation-reduction process, lipid metabolic process. Moreover, since it has been reported that genetic traits are involved in the development of NASH-HCC, we next analyzed the genetic mutations in the STAM™mice. The number of individuals showing mutations in Mtorinvolved in Insulin signaling increases as the disease progresses, especially in the liver cancer phase. These results indicated a clinical correlation of gene profiles in the STAM™mouse.

Keywords: steatosis, non-alcoholic steatohepatitis, fibrosis, hepatocellular carcinoma, RNA-seq

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1991 Native Plants Marketing by Entrepreneurs in the Landscaping Industry in Japan

Authors: Yuki Hara

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Entrepreneurs are welcomed to the landscaping industry, conserving practically and theoretically biological diversity in landscaping construction, although there are limited reports on corporative trials making a market with a new logistics system of native plants (NP) between landscaping companies and nurserymen. This paper explores the entrepreneurial process of a landscaping company, “5byMidori” for NP marketing. This paper employs a case study design. Data are collected in interviews with the manager and designer of 5byMidori, 2 scientists, 1 organization, and 18 nurserymen, fieldworks at two nurseries, observations of marketing activities in three years, and texts from published documents about the business concept and marketing strategy with NP. These data are analyzed by qualitative methods. The results show that NP is suitable for the vision of 5byMidori improving urban desertified environment with closer urban-rural linkage. Professional landscaping team changes a forestry organization into NP producers conserving a large nursery of a mountain. Multifaceted PR based on the entrepreneurial context and personal background of a landscaping venture can foster team members' businesses and help customers and users to understand the biodiversity value of the product. Wider partnerships with existing nurserymen at other sites in many regions need socio-economic incentives and environmental reliability. In conclusion, the entrepreneurial marketing of a landscaping company needs to add more meanings and a variety of merits in terms of ecosystem services, as NP tends to be in academic definition and independent from the cultures like nurseryman and forestry.

Keywords: biological diversity, landscaping industry, marketing, native plants

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1990 Ribotaxa: Combined Approaches for Taxonomic Resolution Down to the Species Level from Metagenomics Data Revealing Novelties

Authors: Oshma Chakoory, Sophie Comtet-Marre, Pierre Peyret

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Metagenomic classifiers are widely used for the taxonomic profiling of metagenomic data and estimation of taxa relative abundance. Small subunit rRNA genes are nowadays a gold standard for the phylogenetic resolution of complex microbial communities, although the power of this marker comes down to its use as full-length. We benchmarked the performance and accuracy of rRNA-specialized versus general-purpose read mappers, reference-targeted assemblers and taxonomic classifiers. We then built a pipeline called RiboTaxa to generate a highly sensitive and specific metataxonomic approach. Using metagenomics data, RiboTaxa gave the best results compared to other tools (Kraken2, Centrifuge (1), METAXA2 (2), PhyloFlash (3)) with precise taxonomic identification and relative abundance description, giving no false positive detection. Using real datasets from various environments (ocean, soil, human gut) and from different approaches (metagenomics and gene capture by hybridization), RiboTaxa revealed microbial novelties not seen by current bioinformatics analysis opening new biological perspectives in human and environmental health. In a study focused on corals’ health involving 20 metagenomic samples (4), an affiliation of prokaryotes was limited to the family level with Endozoicomonadaceae characterising healthy octocoral tissue. RiboTaxa highlighted 2 species of uncultured Endozoicomonas which were dominant in the healthy tissue. Both species belonged to a genus not yet described, opening new research perspectives on corals’ health. Applied to metagenomics data from a study on human gut and extreme longevity (5), RiboTaxa detected the presence of an uncultured archaeon in semi-supercentenarians (aged 105 to 109 years) highlighting an archaeal genus, not yet described, and 3 uncultured species belonging to the Enorma genus that could be species of interest participating in the longevity process. RiboTaxa is user-friendly, rapid, allowing microbiota structure description from any environment and the results can be easily interpreted. This software is freely available at https://github.com/oschakoory/RiboTaxa under the GNU Affero General Public License 3.0.

Keywords: metagenomics profiling, microbial diversity, SSU rRNA genes, full-length phylogenetic marker

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1989 Symbiotic Functioning, Photosynthetic Induction and Characterisation of Rhizobia Associated with Groundnut, Jack Bean and Soybean from Eswatini

Authors: Zanele D. Ngwenya, Mustapha Mohammed, Felix D. Dakora

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Legumes are a major source of biological nitrogen, and therefore play a crucial role in maintaining soil productivity in smallholder agriculture in southern Africa. Through their ability to fix atmospheric nitrogen in root nodules, legumes are a better option for sustainable nitrogen supply in cropping systems than chemical fertilisers. For decades, farmers have been highly receptive to the use of rhizobial inoculants as a source of nitrogen due mainly to the availability of elite rhizobial strains at a much lower compared to chemical fertilisers. To improve the efficiency of the legume-rhizobia symbiosis in African soils would require the use of highly effective rhizobia capable of nodulating a wide range of host plants. This study assessed the morphogenetic diversity, photosynthetic functioning and relative symbiotic effectiveness (RSE) of groundnut, jack bean and soybean microsymbionts in Eswatini soils as a first step to identifying superior isolates for inoculant production. According to the manufacturer's instructions, rhizobial isolates were cultured in yeast-mannitol (YM) broth until the late log phase and the bacterial genomic DNA was extracted using GenElute bacterial genomic DNA kit. The extracted DNA was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and a dendrogram constructed from the band patterns to assess rhizobial diversity. To assess the N2-fixing efficiency of the authenticated rhizobia, photosynthetic rates (A), stomatal conductance (gs), and transpiration rates (E) were measured at flowering for plants inoculated with the test isolates. The plants were then harvested for nodulation assessment and measurement of plant growth as shoot biomass. The results of ERIC-PCR fingerprinting revealed the presence of high genetic diversity among the microsymbionts nodulating each of the three test legumes, with many of them showing less than 70% ERIC-PCR relatedness. The dendrogram generated from ERIC-PCR profiles grouped the groundnut isolates into 5 major clusters, while the jack bean and soybean isolates were grouped into 6 and 7 major clusters, respectively. Furthermore, the isolates also elicited variable nodule number per plant, nodule dry matter, shoot biomass and photosynthetic rates in their respective host plants under glasshouse conditions. Of the groundnut isolates tested, 38% recorded high relative symbiotic effectiveness (RSE >80), while 55% of the jack bean isolates and 93% of the soybean isolates recorded high RSE (>80) compared to the commercial Bradyrhizobium strains. About 13%, 27% and 83% of the top N₂-fixing groundnut, jack bean and soybean isolates, respectively, elicited much higher relative symbiotic efficiency (RSE) than the commercial strain, suggesting their potential for use in inoculant production after field testing. There was a tendency for both low and high N₂-fixing isolates to group together in the dendrogram from ERIC-PCR profiles, which suggests that RSE can differ significantly among closely related microsymbionts.

Keywords: genetic diversity, relative symbiotic effectiveness, inoculant, N₂-fixing

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1988 The Extent of Proliferation, Apoptosis and Angiogenesis at the Site of Injury Determine the Course of Healing Either as Scar Free or as Scarred One in the Appendages of Lizard

Authors: Isha Ranadive, Sonam Patel, Suresh Balakrishnan

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It has been observed that in lizards wound can be healed by either a scar free mechanism or by scarring. The animal model used to study both these healing processes was Northern House Gecko. In lizard, the tail when amputated heals by scar free mechanism which allows it to regenerate, the same is not seen when the limb is amputated. Proliferation, apoptosis, and angiogenesis are the main events which succeed an injury. We observed that proliferation of the cells beneath the wound epidermis was much higher in case of wound healing in tail. This could be because after the wound gets covered by the epithelium, it enters in to a cross-talk with the underlying mesenchyme to recruit a pool of blastemal cells which proliferate and later differentiate to form the lost part through epimorphic regeneration. This was substantiated by mRNA expression levels of various FGFs which facilitate the cross-talk and also by PCNA which is a marker for proliferation. Western blot result reaffirms the same notion. However, in case of the limb, the rate of apoptosis was more than proliferation as there are a lot of debris that needs to be removed. We came to this conclusion as we observed that p53 the apoptotic gene was highly upregulated in case of the scarred tissue. Further, we confirmed this result by checking the anti-apoptotic gene bcl2 and found it to be significantly down-regulated. As we noticed heightened proliferation in the case of scar-free wound healing in tail, angiogenesis was targeted for the study. This is because, when the cells are proliferating they require constant supply of blood and hence neo-vascularization is inevitable. It was observed that the marker of angiogenesis, VEGF, was expressed more during wound healing as compared to the resting stage of tail. Moreover, a high up-regulation was seen in KDR, a receptor of VEGF. Thus, this study reveals how proliferation, apoptosis, and angiogenesis play a key role in the scar-free as well as scarred wound healing.

Keywords: epimorphic regeneration, injury, northern house gecko, wound healing

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1987 Effects of Delphinidin on Lipid Metabolism in HepG2 Cells and Diet-Induced Obese Mice

Authors: Marcela Parra-Vargas, Ana Sandoval-Rodriguez, Roberto Rodriguez-Echevarria, Jose Dominguez-Rosales, Juan Armendariz-Borunda

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Non-alcoholic fatty liver disease (NAFLD) is characterized by an excess of hepatic lipids, and it is to author’s best knowledge, the most prevalent chronic liver disorder. Anthocyanin-rich food consumption is linked to health benefits in metabolic disorders associated with obesity and NAFLD, although the precise functional role of anthocyanidin delphinidin (Dp) has yet to be established. The aim of this study was to investigate the effect of the Dp in NAFLD metabolic alterations by evaluating prevention or amelioration of hepatic lipid accumulation, as well as molecular mechanisms in two experimental obesity-related models of NALFD. In vitro: HepG2 cells were incubated with sodium palmitate (PA, 1 mM) to induce lipotoxic damage, and concomitantly treated with Dp (180 uM) for 24 h. Subsequently, total lipid accumulation was measured by colorimetric staining with Oil Red O, and total intrahepatic triglycerides were determined by an enzymatic assay. To assess molecular mechanisms, cells were pre-treated with PA for 24 h and then exposed to Dp for 1 h. In vivo: four-week-old male C57BL/6Nhsd mice were allocated in two main groups. Mice were fed with standard diet (control) or high-fat and high-carbohydrate diet (45% fat, HFD) for 16 wk to induce NAFLD. Then HFD was divided into subgroups: one treated orally with Dp (15 mg/kg bw, HFD-Dp) every day for 4 wk, while HFD group treated with vehicle (DMSO). Weight and fasting glucose were recorded weekly, while dietary ingestion was measured daily. Insulin tolerance test was performed at the end of treatment. Liver histology was evaluated with H&E and Masson’s trichrome stain. RT-PCR was used to evaluate gene expression and Western Blot to determine levels of protein in both experimental models. Parametric data were analyzed with one-way ANOVA and Tukey’s post-hoc test. Kruskal-Wallis and Mann-Whitney U test for non-parametric data, and P < 0.5 were considered significant. Dp prevented hepatic lipid accumulation by PA in HepG2 hepatocytes. Furthermore, Dp down-regulated gene expression of SREBP1c, FAS, and CPT1a without modifying AMPK phosphorylation levels. In vivo, Dp oral administration did not ameliorate lipid metabolic alterations raised by HFD. Adiposity, dietary ingestion, fasting glucose, and insulin sensitivity after Dp treatment remained similar to HFD group. Histological analysis showed hepatic damage in HFD groups and no differences between HFD and HFD-Dp groups were found. Hepatic gene expression of ACC and FAS were not altered by HFD. SREBP1c was similar in both HFD and HFD-Dp groups. No significant changes were observed in SREBP1c, ACC, and FAS adipose tissue gene expression by HFD or Dp treatment. Additionally, immunoblotting analysis revealed no changes in pathway SIRT1-LKB-AMPK and PPAR alpha by both HFD groups compared to control. In conclusion, the antioxidant Dp may provoke beneficial effects in the prevention of hepatic lipid accumulation. Nevertheless, the oral dose administrated in mice that simulated the total intake of anthocyanins consumed daily by humans has no effect as a treatment on hepatic lipid metabolic alterations and histological abnormalities associated with exposure to chronic HFD. A healthy lifestyle with regular intake of antioxidants such as anthocyanins may prevent metabolic alterations in NAFLD.

Keywords: anthocyanins, antioxidants, delphinidin, non-alcoholic fatty liver disease, obesity

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1986 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

Abstract:

Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

Procedia PDF Downloads 258
1985 Beyond Bindis, Bhajis, Bangles, and Bhangra: Exploring Multiculturalism in Southwest England Primary Schools, Early Research Findings

Authors: Suparna Bagchi

Abstract:

Education as a discipline will probably be shaped by the importance it places on a conceptual, curricular, and pedagogical need to shift the emphasis toward transformative classrooms working for positive change through cultural diversity. Awareness of cultural diversity and race equality has heightened following George Floyd’s killing in the USA in 2020. This increasing awareness is particularly relevant in areas of historically low ethnic diversity which have lately experienced a rise in ethnic minority populations and where inclusive growth is a challenge. This research study aims to explore the perspectives of practitioners, students, and parents towards multiculturalism in four South West England primary schools. A qualitative case study methodology has been adopted framed by sociocultural theory. Data were collected through virtually conducted semi-structured interviews with school practitioners and parents, observation of students’ classroom activities, and documentary analysis of classroom displays. Although one-third of the school population includes ethnically diverse children, BAME (Black, Asian, and Minority Ethnic) characters featured in children's books published in Britain in 2019 were almost invisible, let alone a BAME main character. The Office for Standards in Education, Children's Services and Skills (Ofsted) are vocal about extending the Curriculum beyond the academic and technical arenas for pupils’ broader development and creation of an understanding and appreciation of cultural diversity. However, race equality and community cohesion which could help in the students’ broader development are not Ofsted’s school inspection criteria. The absence of culturally diverse content in the school curriculum highlighted by the 1985 Swann Report and 2007 Ajegbo Report makes England’s National Curriculum look like a Brexit policy three decades before Brexit. A revised National Curriculum may be the starting point with the teachers as curriculum framers playing a significant part. The task design is crucial where teachers can place equal importance on the interwoven elements of “how”, “what” and “why” the task is taught. Teachers need to build confidence in encouraging difficult conversations around racism, fear, indifference, and ignorance breaking the stereotypical barriers, thus helping to create students’ conception of a multicultural Britain. Research showed that trainee teachers in predominantly White areas often exhibit confined perspectives while educating children. Irrespective of the geographical location, school teachers can be equipped with culturally responsive initial and continuous professional development necessary to impart multicultural education. This may aid in the reduction of employees’ unconscious bias. This becomes distinctly pertinent to avoid horrific cases in the future like the recent one in Hackney where a Black teenager was strip-searched during period wrongly suspected of cannabis possession. Early research findings show participants’ eagerness for more ethnic diversity content incorporated in teaching and learning. However, schools are considerably dependent on the knowledge-focused Primary National Curriculum in England. Moreover, they handle issues around the intersectionality of disability, poverty, and gender. Teachers were trained in times when foregrounding ethnicity matters was not happening. Therefore, preoccupied with Curriculum requirements, intersectionality issues, and teacher preparations, schools exhibit an incapacity due to which keeping momentum on ethnic diversity is somewhat endangered.

Keywords: case study, curriculum decolonisation, inclusive education, multiculturalism, qualitative research in Covid19 times

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1984 Breaking the Barriers: Exploring the Barriers to LGBTQ+ Accessing Palliative Care and the Hospice

Authors: Emma Worley, Mhairi De Sainte Croix, Savneet Lochab, Christopher Roberts, Mark Stroud, Mo Salehan, Kevin Jones

Abstract:

Awareness about the importance of teaching about diversity at medical school is growing. In the realm of diversity includes discussion around the LGBTQ+ community. At Bristol, diversity is taught in first or second year. However, echoing and expanding that teaching throughout the curriculum is needed. This feeds into the spiral curriculum but also highlights the relevance of the topic. It is well known that some people in the LGBTQ+ community struggle the access healthcare due to previous negative experiences. In 2019, 1 in 7 LGBTQ+ people avoided seeking medical care due to fears about discrimination. If people have fears about seeking medical help, then seeking help from Palliative care when they are at their most vulnerable situation can be even harder. To improve positive healthcare situations for people who identify as LGBTQ+ needs to start with talking. Along with some of our CTAs (clinical teaching assistants) we created a teaching session to explore the barriers faced by LGBTQ+ and incorporated communication stations into this. Our plan is to run this session as a three-hour session first discussing different topics: ethnical diversity, ‘coming out’, LGBTQ+ in the older generation, transgender. This will be followed by looking more closely at the barriers to accessing the hospice. The next part of the session will encompass two or three communication scenarios hopefully prompting further discussion and reflection on ways to improve our communication. The first scenario outline is a gay man/lesbian woman with lung cancer discussing options around the hospice. The second scenario is a transgender person with female genitalia who now has cervical cancer (as was not followed up on pap smears after the change of name). The third scenario is a HIV homosexual male patient who has been admitted with dementia. He has a partner but is not married. His next of kin is down as his parents but his parents do not know about his sexuality and HIV status. It allows discussion around confidentiality as well as broaching the meaning of ‘family’ in the LGBTQ+ community. We have chosen to pitch this teaching session to Bristol Year 4 students. They will be currently doing their 6-week Palliative care block, which fits in well. Each session will have four students attend. We have been lucky enough to have two CTAs (clinical teaching assistants) who identify as LGBTQ+ offer their experiences and help. They have been able to help us with the preparation and delivery of the session. Given anecdotal evidence and stories helps to highlight the importance and relevance of this session. The aim is to increase awareness of some factors that may contribute to people who identify as LGBTQ+ having a negative healthcare experience. By starting to talk about it allows awareness and only then will we be able to start to change and improve. Our aim, if the sessions run well, is to expand these sessions to different academy hospitals. Therefore, all Bristol 4th year students would have the opportunity to take part in the teaching session. We would like to expand our portfolio of case scenarios, to address so tricker topics such as a transgender person with dementia who reverts back to a different gender. We would also like to recruit a diverse range of actors, ideally people who identify as the patient in the scenario does. For example, a transgender person acts the transgender scenario. This would give authenticity and enhance the student’s learning experience.

Keywords: communication skills, healthcare barriers, LGBTQ+, palliative care

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1983 The Role of Language Strategy on International Survival of Firm: A Conceptual Framework from Resource Dependence Perspective

Authors: Sazzad Hossain Talukder

Abstract:

Survival in the competitive international market with unforeseen environmental contingencies has always been a concern of the firms that led to adopting different strategies to deal with different situations. Language strategy is considered to enhance the international performance of a firm by organizing language diversity and fostering communications within and outside the firm. Yet there is a lack of theoretical attention or model development on the role of language strategy on firm international survival. From resource dependence perspective, the adoption of language strategy and its relationship with firm survival are determined by the firm´s capability to prevent dependency concentration and/or increase relative power on the external environment. However, the impact of language strategy on firm survival is complex and multifaceted as the strategy influence firm performance indirectly through communication, coordination, learning and value creation. The evidence of various types of language strategies and different forms of firm survival also bring in complexities to understand the effects of a language strategy on the international survival of a firm. Based on language literatures and resource dependence logic, certain propositions are developed to conceptualize the relationship between language strategy and firm international survival in this conceptual paper. For the purpose of this paper, a conceptual model is proposed to examine how different kinds of language strategy foster reduction of resource dependency that lead to firm international survival in respond to local responsiveness and global integration. In this proposed model, it is theorized that language strategy has a positive relationship with the international survival of the firm, as the strategy is likely to reduce external resource dependency and increase the ability to continue independent operations both in short and long term.

Keywords: language strategy, language diversity, firm international survival, resource dependence logic

Procedia PDF Downloads 261
1982 Biochemical Characterization of CTX-M-15 from Enterobacter cloacae and Designing a Novel Non-β-Lactam-β-Lactamase Inhibitor

Authors: Mohammad Faheem, M. Tabish Rehman, Mohd Danishuddin, Asad U. Khan

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The worldwide dissemination of CTX-M type β-lactamases is a threat to human health. Previously, we have reported the spread of blaCTX-M-15 gene in different clinical strains of Enterobacteriaceae from the hospital settings of Aligarh in north India. In view of the varying resistance pattern against cephalosporins and other β-lactam antibiotics, we intended to understand the correlation between MICs and catalytic activity of CTX-M-15. In this study, steady-state kinetic parameters and MICs were determined on E. coli DH5α transformed with blaCTX-M-15 gene that was cloned from Enterobacter cloacae (EC-15) strain of clinical background. The effect of conventional β-lactamase inhibitors (clavulanic acid, sulbactam and tazobactam) on CTX-M-15 was also studied. We have found that tazobactam is the best among these inhibitors against CTX-M-15. The inhibition characteristic of tazobactam is defined by its very low IC50 value (6 nM), high affinity (Ki = 0.017 µM) and better acylation efficiency (k+2/K9 = 0.44 µM-1s-1). It forms an acyl-enzyme covalent complex, which is quite stable (k+3 = 0.0057 s-1). Since increasing resistance has been reported against conventional b-lactam antibiotic-inhibitor combinations, we aspire to design a non-b-lactam core containing b-lactamase inhibitor. For this, we screened ZINC database and performed molecular docking to identify a potential non-β-lactam based inhibitor (ZINC03787097). The MICs of cephalosporin antibiotics in combination with this inhibitor gave promising results. Steady-state kinetics and molecular docking studies showed that ZINC03787097 is a reversible inhibitor which binds non-covalently to the active site of the enzyme through hydrogen bonds and hydrophobic interactions. Though, it’s IC50 (180 nM) is much higher than tazobactam, it has good affinity for CTX-M-15 (Ki = 0.388 µM). This study concludes that ZINC03787097 compound can be used as seed molecule to design more efficient non-b-lactam containing b-lactamase inhibitor that could evade pre-existing bacterial resistance mechanisms.

Keywords: ESBL, non-b-lactam-b-lactamase inhibitor, bioinformatics, biomedicine

Procedia PDF Downloads 229
1981 Combining Bio-Molecular and Isotopic Tools to Determine the Fate of Halogenated Compounds in Polluted Groundwater

Authors: N. Balaban, A. Buernstein, F. Gelman, Z. Ronen

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Brominated flame retardants are widespread pollutants, and are known to be toxic, carcinogenic, endocrinic disrupting as well as recalcitrant. The industrial complex Neot Hovav, in the Northern Negev, Israel, is situated above a fractured chalk aquitard, which is polluted by a wide variety of halogenated organic compounds. Two of the abundant pollutants found in the site are Dibromoneopentyl-glycol (DBNPG) and tribromoneopentyl-alcohol (TBNPA). Due to the elusive nature of the groundwater flow, it is difficult to connect between the spatial changes in contaminant concentrations to degradation. In this study, we attempt to determine whether these compounds are biodegraded in the groundwater, and to gain a better understanding concerning the bacterial community in the groundwater. This was achieved through the application of compound-specific isotope analysis (CSIA) of carbon (13^C/12^C) and bromine (81^Br/79^Br), and new-generation MiSeq pyrosequencing. The sampled boreholes were distributed among three main areas of the industrial complex: around the production plant of TBNPA and DBNPG; along the Hovav Wadi (small ephemeral stream) which crosses and drains the industrial complex; and downstream to the industrial area. TBNPA and DBNPG are found in all three areas, with no clear connection to the proximity of the borehole to the production plant. Initial isotopic data of TBNPA from boreholes in the area surrounding the production plant, reveal no changes in the carbon and bromine isotopic values. When observing the microbial groundwater community, the dominant phylum is Proteobacteria. Known anaerobic dehalogenating bacteria such as Dehalococcoides from the Chloroflexi phylum have also been detected. A statistical comparison of the groundwater microbial diversity using a multi-variant ordination of non-metric multidimensional scaling (NMDS) reveals three main clusters in accordance to spatial location in the industrial complex: all the boreholes sampled adjacent to the production plant cluster together and separately from the Wadi Hovav boreholes cluster and the downstream to the industrial area borehole cluster. This work provides the basis for the development and implication of an isotopic fractionation based tool for assessing the biodegradation of brominated organic compounds in contaminated environments, and a novel attempt to characterize the spatial microbial diversity in the contaminated site.

Keywords: biodegradation, brominated flame retardants, groundwater, isotopic fractionation, microbial diversity

Procedia PDF Downloads 225
1980 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

Procedia PDF Downloads 310
1979 Silica Nanoparticles Induced Oxidative Stress and Inflammation in MRC-5 Human Lung Fibroblasts

Authors: Anca Dinischiotu, Sorina Nicoleta Voicu

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Silica nanoparticles (SiO2-NPs) are widely used in consumer products such as paints, plastics, insulation materials, tires, concrete production, as well as in gene delivery systems and imaging procedures. Environmental human exposure to them occurs during utilization of these products, in a time-dependent manner, the uptake being by topic and inhalation route especially. SiO2-NPs enter cells and induce membrane damage, oxidative stress and inflammatory reactions in a concentration-dependent manner. In this study, MRC-5 cells (human fetal lung fibroblasts) were exposed to amorphous SiO2-NPs at a dose of 62.5 μg/ml for 24, 48 and 72 hours. The size distribution of NPs was a lognormal function, in the range 3-14 nm. A time-dependent decrease of total reduced glutathione concentration by 36%, 50%, and 78% and an increase of NO level by 62%, 32%, respectively 24% compared to control were noticed. An up-regulation of NF-kB expression by 20%, 50% respectively 10% and of Nrf-2 by 139%, 58%, and 16% compared to control after 24, 48 and 72 hours was noticed also. The expression of IL-1β, IL-6, IL-8, and COX-2 was up-regulated in a time-dependent manner. Also, the expression of MMP-2 and MMP-9 were down-regulated after 48 and 72 hours, whereas their activities raised in a time-dependent manner. Exposure of cells to NPs up-regulated the expression of inducible NO synthase, as previously was shown, and probably this is the reason for the increased level of NO, that can react with the thiol groups of reduced glutathione molecules, diminishing its concentration Nrf2 is a transcription factor translocated in nucleus, under oxidative stress, where downstream gene expression activates in order to modulate the adaptive intracellular response against oxidative stress. The cross-talk between Nrf2 and NF-kB activities regulates the inflammatory processes. The activation of NF-kB could activate up-regulation of IL-1β, IL-6, and IL-8. The increase of COX-2 expression could be correlated with IL-1β one. Also, probably in response to the pro-inflammatory cytokines, MMP-2 and MMP-9 were induced and activated. In conclusion, the exposure of MRC-5 cells to SiO2-NPs generated inflammation in a time-dependent manner.

Keywords: inflammation, MRC-5 cells, oxidative stress, silica nanoparticles

Procedia PDF Downloads 135
1978 First Report of Rahnella Victoriana Associated with Walnut Decline

Authors: Mohammadreza Hajialigol, Nargues Falahi Charkhabi, Fatemeh Shahryari, Saadat Sarikhani

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BACKGROUND AND OBJECTIVES Iran is the third producer of Persian walnut worldwide. However, its walnut trees have been under threat from decline during last decade. Walnut canker caused by B. nigrifluens and B. rubrifaciens was recorded in multiple regions of Iran. Furthermore, Brenneria rosae subsp. rosae and Gibbsiella quercinecans were recently recognized as responsible for walnut decline in northwestern Iran. This study aimed to identify the causal agent of walnut decline in Kermanshah and Isfahan. MATERIAL AND METHODS Symptomatic samples were collected from affected walnut trees of Kermanshah and Isfahan provinces. The pathogenicity of strains was proved on immature walnut fruits cv. ‘Hartley’ and young green twigs of two-year-old walnut seedling cv. ‘Chandler’. Pathogenic strains were subjected to conventional phenotypic tests. 16S rRNA, gyrB, and infB genes were partially amplified and sequenced. RESULTS Irregular longitudinal cankers and dark lesions were observed in the outer and inner bark, respectively. Twenty-four strains were isolated on EMB-agar media. Fourteen strains were able to cause necrosis and a dark-colored region in the mesocarp and on young green twigs around the inoculation site 14 and 30 days post-inoculation, respectively. Strains were able to hydrolyze Tween 20, Tween 80, gelatin and esculin, however, did not produce indole or urease. Pairwise comparison, the 16S rRNA gene nucleotide sequences of strain I2 were 100% identical with those of Rahnella victoriana FRB 225T. Moreover, a phylogenetic tree reconstructed based on the concatenated sequences of two housekeeping gene fragments, gyrB (601 bp) and infB (615 bp), revealed that the strains I2, I5, and KE6 were clustered with R. victoriana FRB 225T. CONCLUSION To the best of our knowledge, this is the first report of R. victoriana in association with walnut decline. This result is necessary to find resistant genotypes.

Keywords: emerging pathogens, Iran, juglans regia, MLSA

Procedia PDF Downloads 73
1977 Biodegradation of 2,4-Dichlorophenol by Pseudomonas chlororaphis Strain Isolated from Activated Sludge Sample from a Wastewater Treatment Plant in Durban, South Africa

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

Procedia PDF Downloads 241
1976 Effect of Media Osmolarity on Vi Biosynthesis on Salmonella enterica serovar Typhi Strain C6524 Cultured on Batch System

Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

Procedia PDF Downloads 184
1975 Identification of the Key Enzyme of Roseoflavin Biosynthesis

Authors: V. Konjik, J. Schwartz, R. Sandhoff, M. Mack

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The rising number of multi-resistant pathogens demands the development of new antibiotics in order to reduce the lethal risk of infections. Here, we investigate roseoflavin, a vitamin B2 analogue which is produced by Streptomyces davawensis and Streptomyces cinnabarinus. We consider roseoflavin to be a 'Trojan horse' compound. Its chemical structure is very similar to riboflavin but in fact it is a toxin. Furthermore, it is a clever strategy with regard to the delivery of an antibiotic to its site of action but also with regard to the production of this chemical: The producer cell has only to convert a vitamin (which is already present in the cytoplasm) into a vitamin analog. Roseoflavin inhibits the activity of Flavin depending proteins, which makes up to 3.5 % of predicted proteins in organisms sequenced so far. We sequentially knocked out gene clusters and later on single genes in order to find the ones which are involved in the roseoflavin biosynthesis. Consequently, we identified the gene rosB, coding for the protein carrying out the first step of roseoflavin biosynthesis, starting form Flavin mononucleotide. Here we show, that the protein RosB has so far unknown features. It is per se an oxidoreductase, a decarboxylase and an aminotransferase, all rolled into one enzyme. A screen of cofactors revealed needs of oxygen, NAD+, thiamine and glutamic acid to carry out its function. Surprisingly, thiamine is not only needed for the decaboxylation step, but also for the oxidation of 8-demethyl-8-formyl Flavin mononucleotide. We had managed to isolate three different Flavin intermediates with different oxidation states, which gave us a mechanistic insight of RosB functionality. Our work points to a so far new function of thiamine in Streptomyces davawensis. Additionally, RosB could be extremely useful for chemical synthesis. Careful engineering of RosB may allow the site-specific replacement of methyl groups by amino groups in polyaromatic compounds of commercial interest. Finally, the complete clarification of the roseoflavin biosynthesis opens the possibility of engineering cost-effective roseoflavin producing strains.

Keywords: antibiotic, flavin analogue, roseoflavin biosynthesis, vitamin B2

Procedia PDF Downloads 232
1974 Functional Diversity of Pseudomonas: Role in Stimulation of Bean Germination and Common Blight Biocontrol

Authors: Slimane Mokrani, Nabti El hafid

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Description of the subject: Currently, several efforts focus on the study of biodiversity, microbial biotechnology, and the use of ecological strategies. Objectives: The aim of this present work is to determine the functional diversity of bacteria in rhizospheric and non-rhizospheric soils of different plants. Methods: Bacteria were isolated from soil and identified based on physiological and biochemical characters and genotypic taxonomy performed by 16S rDNA and BOX-PCR. As well as the characterization of various PGPR traits. Then, they are tested for their effects on the stimulation of seed germination and the growth of Phaseolus vulgaris L. As well as their biological control activities with regard to the phytopathogenic bacterial isolate Xapf. Results and Discussion: The biochemical and physiological identification of 75 bacterial isolates made it possible to associate them with the two groups of fluorescent Pseudomonas (74.67%) and non-fluorescent Pseudomonas (25.33%). The identification by 16S rDNA of 27 strains made it possible to attribute the majority of the strains to the genus Pseudomonas (81.48%), Serratia (7.41%) and Bacillus (11.11%). The bacterial strains showed a high capacity to produce IAA, siderophores, HCN and to solubilize phosphate. A significant stimulation of germination and growth was observed by applying the Pseudomonas strains. Furthermore, significant reductions in the severity and intensity of the disease caused caused by Xapf were observed. Conclusion: The bacteria described in this present study endowed with different PGPR activities seem to be very promising for their uses as biological control agents and bio-fertilization.

Keywords: biofertilization, biological control, phaseolus vulgaris L, pseudomonas, Xanthomonas axonopodis pv. phaseoli var. fuscans and common blight

Procedia PDF Downloads 71
1973 Conservation of Rare, Endangered and Threaten Medicinal Plants: Participatory Approach

Authors: G. Raviraja Shetty, K. G. Poojitha, Pranay Kumar

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Biodiversity refers to the numbers, variety and variability of living organisms and ecosystem. The climatic and altitudinal variations, coupled with varied ecological habitats of this country, have contributed to the development of immensely rich vegetation with a unique diversity in medicinal plants which provides an important source of medicinal raw materials for traditional medicine systems as well as for pharmaceutical industries in the country and abroad. World Health Organization has listed over 21000 plant species used around the world for medicinal purpose. In India, about 2500 plant species are being used in indigenous system of medicine. The red data book lists 427 Indian Medicinal plant entries on endangered species, of which 28 are considered extinct, 124 endangered, 81 rare, and 34 insufficiently known. It is abundantly clear from the experience of all govt agencies that on their own they cannot efficiently conserve the biodiversity. Participatory Approach with the involvement of local people in conservation is found to be more effective these days. Involvement of local people reduces the cost involved in conservation. Local communities have long tradition of resource use in particular area, hold in depth knowledge and experience of plant which can be invaluable for conservation efforts.Medicinal plants occupy a vital sector of health care system in India and represent a major national resource.There is an immense need for conservation of diversity of medicinal plant wealth for the present and fore coming generations, by adapting the suitable strategy with most appropriate method of conservation.

Keywords: conservation, biodiversity, participatory, medicinal plants

Procedia PDF Downloads 466
1972 Plecoptera Fauna of Alara and Karpuz Streams and Determination of their Relationships with Water Quality

Authors: Hasan Kalyoncu, Ayşe Güneş

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This study was carried on 12 determined stations, on Alara and Karpuz Streams, between January and November 2014. Seasonal samples were taken from the stations to analyze physicochemical parameters and Plecoptera Fauna in the water. The correlation between identified taxa and physicochemical data were tried to determine. As the result of the study, 2088 individuals from Plecoptera fauna were examined, 3 genera and 13 species were identified. The taxa of Brachyptera risi, Capnia bifrons, Dinocras cephalotes, Diura bicaudata, Isogenus nebecula, Isogenus sp., Isoperla grammatica, Leuctra hippopus, Leuctra inermis, Leuctra moselyi, Leuctra sp., Nemoura sp., Perla bipunctata, Perla marginata, Protonemura meyeri and Rhabdiopteryx acuminata were determined. In Alara Stream, the dominant species were; Isogenus nebecula at stations I and IV, Leuctra moselyi at station II, Leuctra hippopus at stations III, V and VI. In Karpuz Stream, Brachyptera risi was the dominant species in all stations. While Leuctra hippopus was the dominant taxon in Alara Stream, in Karpuz Stream it was Brachyptera risi. The highest diversity value was at station III and the lowest was at station VI in Alara Stream and the lowest diversity value was at station VI, while the highest was at station I in Karpuz Stream. In Alara Stream, the most similar stations were I and III, while in Karpuz Stream the highest similarity was determined between stations I and II. As for the evaluation result, the water quality of Alara and Karpuz Streams were determined as at oligosaprobic level.

Keywords: Alara stream, Karpuz stream, plecoptera, water quality

Procedia PDF Downloads 284
1971 A Proposal of Advanced Key Performance Indicators for Assessing Six Performances of Construction Projects

Authors: Wi Sung Yoo, Seung Woo Lee, Youn Kyoung Hur, Sung Hwan Kim

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Large-scale construction projects are continuously increasing, and the need for tools to monitor and evaluate the project success is emphasized. At the construction industry level, there are limitations in deriving performance evaluation factors that reflect the diversity of construction sites and systems that can objectively evaluate and manage performance. Additionally, there are difficulties in integrating structured and unstructured data generated at construction sites and deriving improvements. In this study, we propose the Key Performance Indicators (KPIs) to enable performance evaluation that reflects the increased diversity of construction sites and the unstructured data generated, and present a model for measuring performance by the derived indicators. The comprehensive performance of a unit construction site is assessed based on 6 areas (Time, Cost, Quality, Safety, Environment, Productivity) and 26 indicators. We collect performance indicator information from 30 construction sites that meet legal standards and have been successfully performed. And We apply data augmentation and optimization techniques into establishing measurement standards for each indicator. In other words, the KPI for construction site performance evaluation presented in this study provides standards for evaluating performance in six areas using institutional requirement data and document data. This can be expanded to establish a performance evaluation system considering the scale and type of construction project. Also, they are expected to be used as a comprehensive indicator of the construction industry and used as basic data for tracking competitiveness at the national level and establishing policies.

Keywords: key performance indicator, performance measurement, structured and unstructured data, data augmentation

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1970 Tip60’s Novel RNA-Binding Function Modulates Alternative Splicing of Pre-mRNA Targets Implicated in Alzheimer’s Disease

Authors: Felice Elefant, Akanksha Bhatnaghar, Keegan Krick, Elizabeth Heller

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Context: The severity of Alzheimer’s Disease (AD) progression involves an interplay of genetics, age, and environmental factors orchestrated by histone acetyltransferase (HAT) mediated neuroepigenetic mechanisms. While disruption of Tip60 HAT action in neural gene control is implicated in AD, alternative mechanisms underlying Tip60 function remain unexplored. Altered RNA splicing has recently been highlighted as a widespread hallmark in the AD transcriptome that is implicated in the disease. Research Aim: The aim of this study was to identify a novel RNA binding/splicing function for Tip60 in human hippocampus and impaired in brains from AD fly models and AD patients. Methodology/Analysis: The authors used RNA immunoprecipitation using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. To identify Tip60’s RNA targets, they performed genome sequencing (DNB-SequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Findings: The authors' transcriptomic analysis of RNA bound to Tip60 by Tip60-RNA immunoprecipitation (RIP) revealed Tip60 RNA targets enriched for critical neuronal processes implicated in AD. Remarkably, 79% of Tip60’s RNA targets overlap with its chromatin gene targets, supporting a model by which Tip60 orchestrates bi-level transcriptional regulation at both the chromatin and RNA level, a function unprecedented for any HAT to date. Since RNA splicing occurs co-transcriptionally and splicing defects are implicated in AD, the authors investigated whether Tip60-RNA targeting modulates splicing decisions and if this function is altered in AD. Replicate multivariate analysis of transcript splicing (rMATS) analysis of RNA-Seq data sets from wild-type and AD fly brains revealed a multitude of mammalian-like AS defects. Strikingly, over half of these altered RNAs were bonafide Tip60-RNA targets enriched for in the AD-gene curated database, with some AS alterations prevented against by increasing Tip60 in fly brain. Importantly, human orthologs of several Tip60-modulated spliced genes in Drosophila are well characterized aberrantly spliced genes in human AD brains, implicating disruption of Tip60’s splicing function in AD pathogenesis. Theoretical Importance: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology. Data Collection: The authors collected data from RNA immunoprecipitation experiments using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. They also performed genome sequencing (DNBSequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Questions: The question addressed by this study was whether Tip60 has a novel RNA binding/splicing function in human hippocampus and whether this function is impaired in brains from AD fly models and AD patients. Conclusions: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology.

Keywords: Alzheimer's disease, cognition, aging, neuroepigenetics

Procedia PDF Downloads 64
1969 Estimating Understory Species Diversity of West Timor Tropical Savanna, Indonesia: The Basis for Planning an Integrated Management of Agricultural and Environmental Weeds and Invasive Species

Authors: M. L. Gaol, I. W. Mudita

Abstract:

Indonesia is well known as a country covered by lush tropical rain forests, but in fact, the northeastern part of the country, within the areas geologically known as Lesser Sunda, the dominant vegetation is tropical savanna. Lesser Sunda is a chain of islands located closer to Australia than to islands in the other parts of the country. Among those of islands in the chain which is closes to Australia, and thereby most strongly affected by the hot and dry Australian climate, is the island of Timor, the western part of which belongs to Indonesia and the eastern part is a sovereign state East Timor. Regardless of being the most dominant vegetation cover, tropical savanna in West Timor, especially its understory, is rarely investigated. This research was therefore carried out to investigate the structure, composition and diversity of the understory of this tropical savanna as the basis for looking at the possibility of introducing other spesieis for various purposes. For this research, 14 terrestrial communities representing major types of the existing savannas in West Timor was selected with aid of the most recently available satellite imagery. At each community, one stand of the size of 50 m x 50 m most likely representing the community was as the site of observation for the type of savanna under investigation. At each of the 14 communities, 20 plots of 1 m x 1 m in size was placed at random to identify understory species and to count the total number of individuals and to estimate the cover of each species. Based on such counts and estimation, the important value of each species was later calculated. The results of this research indicated that the understory of savanna in West Timor consisted of 73 understory species. Of this number of species, 18 species are grasses and 55 are non-grasses. Although lower than non-grass species, grass species indeed dominated the savanna as indicated by their number of individuals (65.33 vs 34.67%), species cover (57.80 vs 42.20%), and important value (123.15 vs 76.85). Of the 14 communities, the lowest density of grass was 13.50/m2 and the highest was 417.50/m2. Of 18 grass species found, all were commonly found as agricultural weeds, whereas of 55 non-grass, 10 species were commonly found as agricultural weeds, environmental weeds, or invasive species. In terms of better managing the savanna in the region, these findings provided the basis for planning a more integrated approach in managing such agricultural and environmental weeds as well as invasive species by considering the structure, composition, and species diversity of the understory species existing in each site. These findings also provided the basis for better understanding the flora of the region as a whole and for developing a flora database of West Timor in future.

Keywords: tropical savanna, understory species, integrated management, weedy and invasive species

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1968 Identification of Blood Biomarkers Unveiling Early Alzheimer's Disease Diagnosis Through Single-Cell RNA Sequencing Data and Autoencoders

Authors: Hediyeh Talebi, Shokoofeh Ghiam, Changiz Eslahchi

Abstract:

Traditionally, Alzheimer’s disease research has focused on genes with significant fold changes, potentially neglecting subtle but biologically important alterations. Our study introduces an integrative approach that highlights genes crucial to underlying biological processes, regardless of their fold change magnitude. Alzheimer's Single-cell RNA-seq data related to the peripheral blood mononuclear cells (PBMC) was extracted from the Gene Expression Omnibus (GEO). After quality control, normalization, scaling, batch effect correction, and clustering, differentially expressed genes (DEGs) were identified with adjusted p-values less than 0.05. These DEGs were categorized based on cell-type, resulting in four datasets, each corresponding to a distinct cell type. To distinguish between cells from healthy individuals and those with Alzheimer's, an adversarial autoencoder with a classifier was employed. This allowed for the separation of healthy and diseased samples. To identify the most influential genes in this classification, the weight matrices in the network, which includes the encoder and classifier components, were multiplied, and focused on the top 20 genes. The analysis revealed that while some of these genes exhibit a high fold change, others do not. These genes, which may be overlooked by previous methods due to their low fold change, were shown to be significant in our study. The findings highlight the critical role of genes with subtle alterations in diagnosing Alzheimer's disease, a facet frequently overlooked by conventional methods. These genes demonstrate remarkable discriminatory power, underscoring the need to integrate biological relevance with statistical measures in gene prioritization. This integrative approach enhances our understanding of the molecular mechanisms in Alzheimer’s disease and provides a promising direction for identifying potential therapeutic targets.

Keywords: alzheimer's disease, single-cell RNA-seq, neural networks, blood biomarkers

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