Search results for: Asiatic wild elephant

Authors: Konstantin S. Vainutis, Mark E. Andreev, Anastasia N. Voronova, Mikhail Yu. Shchelkanov

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Introduction: The trematodes from the family Cyclocoelidae (cyclocoelids) belong to the superfamily Echinostomatoidea infecting air sacs and trachea of wild birds. At present, the family Cyclocoelidae comprises nine valid genera in three subfamilies: Cyclocoelinae (type taxon), Haematotrephinae, and Typhlocoelinae. To our best knowledge, in this study, molecular genetic methods were used for the first time for studying cyclocoelids from the Russian Far East. Here we provide the data on the morphology and phylogeny of cyclocoelids from gadwall from the Russian Far East. The morphological and genetic data obtained for cyclocoelids indicated the necessity to revise the previously proposed classification within the family Cyclocoelidae. Objectives: The first objective was performing the morphological study of cyclocoelids found in M. strepera from the Russian Far East. The second objective is to reconstruct the phylogenetic relationships of the studied trematodes with other cyclocoelids using the 28S gene. Material and methods: During the field studies in the Khasansky district of the Primorsky region, 21 cyclocoelids were recovered from the air sacs of a single gadwall Mareca strepera. Seven samples of cyclocoelids were overstained in alum carmine, dehydrated in a graded ethanol series, cleared in clove oil, and mounted in Canada balsam. Genomic DNA was extracted from four cyclocoelids using the alkaline lysis method HotShot. The 28S rDNA fragment was amplified using the forward primer Digl2 and the reverse primer 1500R. Results: According to morphological features (ovary intratesticular, forming a triangle with the testes), the studied worms belong to the subfamily Cyclocoelinae Stossich, 1902. In particular, the highest morphological similarity was observed in relation to the trematodes of the genus Cyclocoelum Brandes, 1892 – genital pores are pharyngeal. However, the genetic analysis has shown significant discrepancies between the trematodes studied regarding the genus Cyclocoelum. On the phylogenetic tree, these trematodes took the sister position in relation to the genus Morishitium (previously considered in the subfamily Szidatitrematinae). Conclusion: Based on the results of the morphological and genetic studies, cyclocoelids isolated from Mareca strepera are suggested to be described in the previously unknown genus and differentiated from the type genus Cyclocoelum of the type subfamily Cyclocoelinae. Considering the available molecular data, including described cyclocoelids, the family Cyclocoelidae comprises ten valid genera in the three subfamilies mentioned above.

Keywords: new species, trematoda, phylogeny, cyclocoelidae

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Authors: N. Mahon, I. Crute, M. Di Bonito, E. Simmons, M. M. Islam

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Growing interest in the concept of the sustainable intensification (SI) of agriculture has been shown by, national governments, transnational agribusinesses, intergovernmental organizations and research institutes, amongst others. This interest may be because SI is seen as a ‘third way’ for agricultural development, between the seemingly disparate paradigms of ‘intensive’ agriculture and more ‘sustainable’ forms of agriculture. However, there is a lack of consensus as to what SI means in practice and how it should be measured using indicators of change. This has led to growing confusion, disagreement and skepticism regarding the concept, especially amongst civil society organizations, both in the UK and other countries. This has prompted the need for bottom-up, participatory approaches to identify indicators of SI. Our aim is to identify the views of British stakeholders regarding the areas of agreement and disagreement as to what SI is and how it should be measured in the UK using indicators of change. Data for this investigation came from 32 semi-structured interviews, conducted between 2015 and 2016, with stakeholders from throughout the UK food system. In total 110 indicators of SI were identified. These indicators covered a wide variety of subjects including biophysical, social and political considerations. A number of indicators appeared to be widely applicable and were similar to those suggested in the global literature. These include indicators related to the management of the natural resources on which agriculture relies e.g., ‘Soil organic matter’, ‘Number of pollinators per hectare’ and ‘Depth of water table’. As well as those related to agricultural externalities, e.g., ‘Greenhouse gas emissions’ and ‘Concentrations of agro-chemicals in waterways’. However, many of the indicators were much more specific to the context of the UK. These included, ‘Areas of high nature value farmland’, ‘Length of hedgerows per hectare’ and ‘Age of farmers’. Furthermore, tensions could be seen when participants considered the relative importance of agricultural mechanization versus levels of agricultural employment, the pros and cons of intensive, housed livestock systems and value of wild biodiversity versus the desire to increase agricultural yields. These areas of disagreement suggest the need to carefully consider the trade-offs inherent in the concept. Our findings indicate that in order to begin to resolve the confusions surrounding SI it needs to be considered in a context specific manner, rather than as a single uniform concept. Furthermore, both the environmental and the social parameters in which agriculture operates need to be considered in order to operationalize SI in a meaningful way. We suggest that participatory approaches are key to this process, facilitating dialogue and collaborative-learning between all the stakeholders, allowing them to reach a shared vision for the future of agricultural development.

Keywords: agriculture, indicators, participatory approach, sustainable intensification

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Authors: Marcus Papandrea

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Gerace survives as one of the best examples of unspoiled Byzantine heritage in Calabria and the world due to its strategic location. As the last western province of the Byzantine Empire, Calabria was not subject to the destruction or conversion of sites which took place by the Ottomans in the east or the Arabs in Sicily and North Africa. Situated ten kilometers inland atop a 500m high table mountain, Gerace overlooks the Ionian coast and is a gateway to the rugged and wild mountain interior of the Calabrian peninsula. It is only connected to the outside world by a single windy and crumbling road and, unfortunately, faces serious economic and demographic decline. Largely due to its isolation, Gerace has remained understudied and under-recognized in a country that boasts the most UNESCO sites in the world despite its wealth and high density of Byzantine monuments. In 1995, the Patriarch of the Eastern Orthodox church, Bartholomew I, visited Gerace. He re-opened and blessed the ancient Byzantine church San Giovanni Crisostomo, reviving Gerace’s cultural origins and links to Byzantium. This paper examines how these links have persisted over a millennium, starting from the community’s humble origins as a refuge for ascetic monks to becoming the “city of one-hundred churches.” While little is documented or written about Gerace’s early history, this paper employs archaeological findings as well as hagiography to present valuable insight into this area which became known as the “land of the saints.” By characterizingGerace’s early Byzantine society and helping to understand its strong spiritual roots, this paper creates the basis necessary to understand the endurance of its Byzantine legacy and appreciate its important cultural contributions to the Italian Renaissance as a hub of Greek literacy which attracted great humanists from the fourteenth to fifteenth century such as Barlaam of Seminara, Simone Autumano, Bessarion, and AthanasioChalkeolopus.Inbringing together these characters, this paper propels Gerace onto the world stage as an important cultural center in medieval Mediterranean history which facilitated cross cultural interactions between Byzantine Greeks, Sicilian Arabs, Jews, and Normans. From this intersection developed a syncretism which led to modern-day Calabrian identity culture and society and is perhaps most visible in some of Gerace’s last surviving monuments from this time. While emphasizing this unassuming town’s cultural importance and unique Byzantine heritage, this paper also highlights the criteria which Gerace fulfills for being included in the World Heritage List.

Keywords: byzantine rite, greek rite, italo-greek, latinization

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Authors: Khasim Munir Bhasha Shaik

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Veligonda and adjoining hill range spread along about 170 Km North to South in Kadapa and Nellore Districts stretching a little further into Prakasam District. The latitude in general ranges up to 1000m. The forests are generally dry deciduous type. Veligonda and adjoining hill ranges comprise of Palakonda, Seshachalam, Lankamala and the terminal part of Nallamalais from mid-region of Southern Eastern Ghats. The Veligonda range which separates the Nellore district from Kadapa and Kurnool is the backbone of the Eastern Ghats, starting from Nagari promontory in Chittoor district. It runs in a northerly direction along the western border of the Nellore district, with a raising elevation of 3,626 ft at Penchalakona in Raipur thaluk. Veligonda hill ranges are high in altitude and have deep valleys. Among the Veligondas range of hills the Durgam in Venkatagiri range and Penchalakona are the most prominent and are situated 914 meters above mean sea level. It has more than 3000 species of plants along with 500 animal species. The unique specialty of this region is the presence of Pterocarpus santalinus(endangered) and Santalum album (vulnerable). In the present study, an attempt is made to assess the efforts that are going on to conserve the biodiversity of flora and fauna of this region. Various conservation strategies were suggested to protect the biodiversity and richness of Veligonda forest, hill region of Eastern Ghats of Andhra Pradesh. The major threats and the reasons for the dwindling species richness are poor rainfall, adverse climatic conditions, robbery of Red sanders and poaching of animals by the local tribals. Efforts are to be made to conserve some of the animals by both in situ and ex-situ methods. More awareness is to be developed among the local communities who are dwelling in the vicinity and importance of conservation is to be emphasized to them. Anthropogenic attachments are to be made by introducing more numbers of sacred groves. Gross enforcement of law is to be made to protect the various forest resources in this area. The important species with the medicinal values are to be identified. It was found that two important wildlife sanctuaries named Sri Lankamalleswarawildlife sanctuary and Sripenusila Narasimha wildlife sanctuary are working for the comprehensive conservation of the environment in this area. Apart from this more than 38 important sacred grooves are there where the plants and animals are protected by local Yanadi and other communities.

Keywords: biodiversity, wild life sanctuary, habitat destruction, eastern Ghats

Procedia PDF Downloads 155

Authors: Arlene López-Sampson, Tony Page, Betsy Jackes

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Aquilaria genus produces a highly valuable fragrant oleoresin known as agarwood. Agarwood forms in a few trees in the wild as a response to injure or pathogen attack. The resin is used in perfume and incense industry and medicine. Cultivation of Aquilaria species as a sustainable source of the resin is now a common strategy. Physiological traits are frequently used as a proxy of crop and tree productivity. Aquilaria species growing in Queensland, Australia were studied to investigate relationship between leaf-productivity traits with tree growth. Specifically, 28 trees, representing 12 plus trees and 16 trees from yield plots, were selected to conduct carbon isotope analysis (δ13C) and monitor six leaf attributes. Trees were grouped on four diametric classes (diameter at 150 mm above ground level) ensuring the variability in growth of the whole population was sampled. Model averaging technique based on the Akaike’s information criterion (AIC) was computed to identify whether leaf traits could assist in diameter prediction. Carbon isotope values were correlated with height classes and leaf traits to determine any relationship. In average four leaves per shoot were recorded. Approximately one new leaf per week is produced by a shoot. Rate of leaf expansion was estimated in 1.45 mm day-1. There were no statistical differences between diametric classes and leaf expansion rate and number of new leaves per week (p > 0.05). Range of δ13C values in leaves of Aquilaria species was from -25.5 ‰ to -31 ‰ with an average of -28.4 ‰ (± 1.5 ‰). Only 39% of the variability in height can be explained by δ13C in leaf. Leaf δ13C and nitrogen content values were positively correlated. This relationship implies that leaves with higher photosynthetic capacities also had lower intercellular carbon dioxide concentrations (ci/ca) and less depleted values of 13C. Most of the predictor variables have a weak correlation with diameter (D). However, analysis of the 95% confidence of best-ranked regression models indicated that the predictors that could likely explain growth in Aquilaria species are petiole length (PeLen), values of δ13C (true13C) and δ15N (true15N), leaf area (LA), specific leaf area (SLA) and number of new leaf produced per week (NL.week). The model constructed with PeLen, true13C, true15N, LA, SLA and NL.week could explain 45% (R2 0.4573) of the variability in D. The leaf traits studied gave a better understanding of the leaf attributes that could assist in the selection of high-productivity trees in Aquilaria.

Keywords: 13C, petiole length, specific leaf area, tree growth

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Authors: Mina Rabipour, Elena Pallaske, Floyd Hassenrück, Rocio Rebollido-Rios

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Protein tyrosine kinases, including Lyn kinase of the Src family kinases (SFK), regulate cell proliferation, survival, and differentiation. Lyn kinase has been implicated in various cancers, positioning it as a promising therapeutic target. However, the conserved ATP-binding pocket across SFKs makes developing selective inhibitors challenging. This study aims to address this limitation by exploring the potential for allosteric modulation of Lyn kinase, focusing on how its structural dynamics and specific oncogenic mutations impact its conformation and function. To achieve this, we combined homology modeling, molecular dynamics simulations, and data science techniques to conduct microsecond-length simulations. Our approach allowed a detailed investigation into the interplay between Lyn’s catalytic and regulatory domains, identifying key conformational states involved in allosteric regulation. Additionally, we evaluated the structural effects of Dasatinib, a competitive inhibitor, and ATP binding on Lyn active conformation. Notably, our simulations show that cancer-related mutations, specifically I364L/N and E290D/K, shift Lyn toward an inactive conformation, contrasting with the active state of the wild-type protein. This may suggest how these mutations contribute to aberrant signaling in cancer cells. We conducted a dynamical network analysis to assess residue-residue interactions and the impact of mutations on the Lyn intramolecular network. This revealed significant disruptions due to mutations, especially in regions distant from the ATP-binding site. These disruptions suggest potential allosteric sites as therapeutic targets, offering an alternative strategy for Lyn inhibition with higher specificity and fewer off-target effects compared to ATP-competitive inhibitors. Our findings provide insights into Lyn kinase regulation and highlight allosteric sites as avenues for selective drug development. Targeting these sites may modulate Lyn activity in cancer cells, reducing toxicity and improving outcomes. Furthermore, our computational strategy offers a scalable approach for analyzing other SFK members or kinases with similar properties, facilitating the discovery of selective allosteric modulators and contributing to precise cancer therapies.

Keywords: lyn tyrosine kinase, mutation analysis, conformational changes, dynamic network analysis, allosteric modulation, targeted inhibition

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Authors: Ratna Sharma, Kaveri Dutta

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Assam (India), the gate way to the Northeast India is mainly known for its exquisite silks, the art and craft. The state has a rich collection of traditional jewellery which is unique and exclusive to the state. These jewelleries hold a special place in the heart of the Assamese women. Similarly handloom industry of Assam is basically silk oriented. Among the wild silk, Eri silk fabric has remained as “the poor man’s silk” but it is closely attached to the assamese society, dress for it's warm quality. In view of the changing market trends, fashion and consumer demands, Silk is emerging as a fashion fabric both in India and abroad. In case of Eri silk fabric it has limited use in clothing and accessories. Hence the restructured and redesigned traditional jewellery motifs of Assam (India) over Eri silk products will have greater potential in reviving the decline of art, generate revenue, self employment towards craftsmen and also recognition of the art. The information incorporated in the paper is primary and the data have been collected by purposive sampling method. This work of art was expressed on Eri silk fabric in the form of traditional hand embroidery as it is closely connected with the era of the individual in history of mankind and reflects the personal expression of an entity. For this study selected traditional motifs of Assamese ornaments was used. Some of the popular traditional Assamese jewellery include earrings with exquisite Lokaparo, Keru, Thuriya, Jangphai, etc. An array of necklaces including Golpata, Satsori, Jon biri, Bena, Gejera, Dhol biri, Doog doogi, Biri Moni, Mukuta Moni, Poalmoni, Silikha Moni and Magardana and diversified rings including Senpata, Horinsakua, Jethinejia, bakharpata and others. Selected two motifs each from necklace, earring and finger ring designs. Selected motifs were further developed into 3 categories- the border, the main motif and all over butta followed by placement of developed patterns on products. Products developed were stoles, scarf’s, purses, brooch pins, skirts for women and ties, handkerchief, jackets for men. The developed products were surveyed by selected respondents. From the present study it can be observed that the embellished traditional jewellery motifs resulted in fresh and colourful pattern on developed Eri silk products. Moreover the motifs which were gradually fading among the community itself showed a very good recognition towards art. The embroidered Eri silk fabric also created a huge change in a positive way among craftsman.

Keywords: Art and craft of Assam, eri silk, hand embroidery, traditional Assamese jewellery motifs

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Authors: Shraddha Rane, Richard Warren, Stephen Eyre

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L-23 is a pro-inflammatory molecule that signals T cells to release cytokines such as IL-17A and IL-22. Psoriasis is driven by a dysregulated immune response, within which IL-23 is now thought to play a key role. Genome-wide association studies (GWAS) have identified a number of genetic risk loci that support the involvement of IL-23 signalling in psoriasis; in particular a robust susceptibility locus at a gene encoding a subunit of the IL-23 receptor (IL23R) (Stuart et al., 2015; Tsoi et al., 2012). The lead psoriasis-associated SNP rs9988642 is located approximately 500 bp downstream of IL23R but is in tight linkage disequilibrium (LD) with a missense SNP rs11209026 (R381Q) within IL23R (r2 = 0.85). The minor (G) allele of rs11209026 is present in approximately 7% of the population and is protective for psoriasis and several other autoimmune diseases including IBD, ankylosing spondylitis, RA and asthma. The psoriasis-associated missense SNP R381Q causes an arginine to glutamine substitution in a region of the IL23R protein between the transmembrane domain and the putative JAK2 binding site in the cytoplasmic portion. This substitution is expected to affect the receptor’s surface localisation or signalling ability, rather than IL23R expression. Recent studies have also identified a psoriatic arthritis (PsA)-specific signal at IL23R; thought to be independent from the psoriasis association (Bowes et al., 2015; Budu-Aggrey et al., 2016). The lead PsA-associated SNP rs12044149 is intronic to IL23R and is in LD with likely causal SNPs intersecting promoter and enhancer marks in memory CD8+ T cells (Budu-Aggrey et al., 2016). It is therefore likely that the PsA-specific SNPs affect IL23R function via a different mechanism compared with the psoriasis-specific SNPs. It could be hypothesised that the risk allele for PsA located within the IL23R promoter causes an increase IL23R expression, relative to the protective allele. An increased expression of IL23R might then lead to an exaggerated immune response. The independent genetic signals identified for psoriasis and PsA in this locus indicate that different mechanisms underlie these two conditions; although likely both affecting the function of IL23R. It is very important to further characterise these mechanisms in order to better understand how the IL-23 receptor and its downstream signalling is affected in both diseases. This will help to determine how psoriasis and PsA patients might differentially respond to therapies, particularly IL-23 biologics. To investigate this further we have developed an in vitro model using CD4 T cells which express either wild type IL23R and IL12Rβ1 or mutant IL23R (R381Q) and IL12Rβ1. Model expressing different isotypes of IL23R is also underway to investigate the effects on IL23R expression. We propose to further investigate the variants for Ps and PsA and characterise key intracellular processes related to the variants.

Keywords: IL23R, psoriasis, psoriatic arthritis, SNP

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Authors: Krzysztof Buczak, Sonia Lachowska, Pawel Kucharski, Agnieszka Antonczyk

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Tiletamine - zolazepam combination is increasingly used in veterinary anaesthesiology in wild animals, including snakes. The available literature shows a lack of information about anesthesia in this mixture in ovoviviparous snakes. The studies show the possibility of using the combination at a dose of 20 mg/kg or more for snake immobilization. This paper presents an anesthetic protocol with the use of a combination of tiletamine - zolazepam at the dose of 10 mg/kg intramuscularly and maintenance with inhalant anesthesia with isoflurane in pure oxygen. The objective of this study was to evaluate the usefulness of the anesthetic protocol to proceed with coeliotomy in Amazon Tree Boa. The patient was a five years old bicolor female Amazon Tree Boa (Corallus hortulanus) with dystocia. The clinical examination reveals significant emaciation (bodyweight 520g), high degree of dehydration, heart rate (HR = 60 / min), pale mucous membranes and poor reactivity. Meloxicam (1 mg/kg) and tramadol (10 mg/kg) were administered subcutaneously and the patient was placed in an incubator with access to fresh oxygen. Four hours later, the combination of tiletamine - zolazepam (10 mg/kg) was administered intramuscularly for induction of anesthesia. The snake was intubated and connected to inhalant anesthesia equipment. For maintenance, the anesthesia isoflurane in pure oxygen was used due to apnea, which occurs 30 minutes after the induction semi-closed system was attached and the ventilator was turned on (PCV system, four breaths per minute, 8 cm of H2O). Cardiopulmonary parameters (HR, RR, SPO2, ETCO2, ETISO) were assessed throughout the procedure. During the entire procedure, the operating room was heated to a temperature of 26 degrees Celsius. Additionally, the hose was placed on a heating mat, which maintained a temperature of 30 degrees Celsius. For 15 minutes after induction, the loss of muscle tone was observed from the head to the tail. Induction of general anesthesia was scored as good because of the possibility of intubation. During the whole procedure, the heart rate was at the rate of 58 beats per minute (bpm). Ventilation parameters were stable throughout the procedure. The recovery period lasts for about 4 hours after the end of general anesthesia. The muscle tension returned from tail to head. The snake started to breathe spontaneously within 1,5 hours after the end of general anesthesia. The protocol of general anesthesia with the combination of tiletamine- zolazepam with a dose of 10 mg/kg is useful for proceeding with the emergency coeliotomy in maintenance with isoflurane in oxygen. Further study about the impact of the combination of tiletamine- zolazepam for the recovery period is needed.

Keywords: anesthesia, corallus hortulanus, ovoviparous, snake, tiletamine, zolazepam

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Authors: Mirjana Petronijević, Sanja Panić, Aleksandra Cvetanović, Branko Kordić, Nenad Grba

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Enzyme peroxidases are biological catalysts and play a major role in phenolic wastewater treatments and other environmental applications. The most studied species from the peroxidases family is horseradish peroxidase (HRP). In environmental processes, HRP could be used in its free or immobilized form. Enzyme immobilization onto solid support is performed to improve the enzyme properties, prolong its lifespan and operational stability and allow its reuse in industrial applications. One of the enzyme supports of a newer generation is magnetic particles (MPs). Fe₃O₄ MPs are the most widely pursued immobilization of enzymes owing to their remarkable advantages of biocompatibility and non-toxicity. Also, MPs can be easily separated and recovered from the water by applying an external magnetic field. On the other hand, metals and metal oxides are not suitable for the covalent binding of enzymes, so it is necessary to perform their surface modification. Fe₃O₄ MPs functionalization could be performed during the process of their synthesis if it takes place in the presence of plant extracts. Extracts of plant material, such as wild plants, herbs, even waste materials of the food and agricultural industry (bark, shell, leaves, peel), are rich in various bioactive components such as polyphenols, flavonoids, sugars, etc. When the synthesis of magnetite is performed in the presence of plant extracts, bioactive components are incorporated into the surface of the magnetite, thereby affecting its functionalization. In this paper, the suitability of bio-magnetite as solid support for covalent immobilization of HRP across glutaraldehyde was examined. The activity of immobilized HRP at different pH values (4-9) and temperatures (20-80°C) and reusability were examined. Bio-MP was synthesized by co-precipitation method from Fe(II) and Fe(III) sulfate salts in the presence of water extract of the Allium cepa peel. The water extract showed 81% of antiradical potential (according to DPPH assay), which is connected with the high content of polyphenols. According to the FTIR analysis, the bio-magnetite contains oxygen functional groups (-OH, -COOH, C=O) suitable for binding to glutaraldehyde, after which the enzyme is covalently immobilized. The immobilized enzyme showed high activity at ambient temperature and pH 7 (30 U/g) and retained ≥ 80% of its activity at a wide range of pH (5-8) and temperature (20-50°C). The HRP immobilized onto bio-MPs showed remarkable stability towards temperature and pH variations compared to the free enzyme form. On the other hand, immobilized HRP showed low reusability after the first washing cycle enzyme retains 50% of its activity, while after the third washing cycle retains only 22%.

Keywords: bio-magnetite, enzyme immobilization, water extracts, environmental protection

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Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

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Authors: Bagam Venkateswarlu

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NMDC Limited, Indian multinational mining company operates under administrative control of Ministry of Steel, Government of India. This study is undertaken to evaluate how sustainable mining practiced by the company fulfils the provisions of Indian Constitution to secure to its citizen – justice, equality of status and opportunity, promoting social, economic, political, and religious wellbeing. The Constitution of India lays down a road map as to how the goal of being a “Welfare State” shall be achieved. The vision of sustainable mining being practiced is oriented along the constitutional responsibilities on Indian Citizens and the Corporate World. This qualitative study shall be backed by quantitative studies of National Mineral Development Corporation performances in various domains of sustainable mining and ESG, that is, environment, social and governance parameters. For example, Five Star Rating of mine is a comprehensive evaluation system introduced by Ministry of Mines, Govt. of India is one of the methodologies. Corporate Social Responsibilities is one of the thrust areas for securing social well-being. Green energy initiatives in and around the mines has given the title of “Eco-Friendly Miner” to NMDC Limited. While operating fully mechanized large scale iron ore mine (18.8 million tonne per annum capacity) in Bacheli, Chhattisgarh, M/s NMDC Limited caters to the needs of mineral security of State of Chhattisgarh and Indian Union. It preserves forest, wild-life, and environment heritage of richly endowed State of Chhattisgarh. In the remote and far-flung interiors of Chhattisgarh, NMDC empowers the local population by providing world class educational & medical facilities, transportation network, drinking water facilities, irrigational agricultural supports, employment opportunities, establishing religious harmony. All this ultimately results in empowered, educated, and improved awareness in population. Thus, the basic tenets of constitution of India- secularism, democracy, welfare for all, socialism, humanism, decentralization, liberalism, mixed economy, and non-violence is fulfilled. Constitution declares India as a welfare state – for the people, of the people and by the people. The sustainable mining practices by NMDC are in line with the objective. Thus, the purpose of study is fully met with. The potential benefit of the study includes replicating this model in existing or new establishments in various parts of country – especially in the under-privileged interiors and far-flung areas which are yet to see the lights of development.

Keywords: ESG values, Indian constitution, NMDC limited, sustainable mining, CSR, green energy

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Authors: Dace Grauda, Gunta Cekstere, Inta Belogrudova, Andis Karlsons, Isaak Rashal

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Trifolium repens L. (white or Dutch clover) is a perennial herb, belongs to legume family (Leguminosae Juss.), spread extensively by stolons and seeds. The species is cultivated worldwide and was naturalized in many countries in meadows, yards, gardens, along roads and streets etc., especially in temperate regions. It is widespread also in grasslands throughout Riga, the capital of Latvia. The goal of this study was to investigate genetic structure of white clover population in Riga and to evaluate influence of different salt concentration on plants. For this purpose universal retrotranspozone based IRAP (Inter-Retrotransposon Amplified Polymorphism) method was used. The plant material was collected in different regions of Riga and in several urban areas of Latvia. Plant DNA was isolated from in silicogel dried leaves of using 1% CTAB (cetyltrimet-ammonium bromide) buffer DNA extraction procedure. Genetic structure of city population and wild populations were compared. Soil salinization is an important issue associated with low water resources and highly urbanized areas in aride and semi-aride climate conditions, as well as de-icing salt application to prevent ice formation on roads in winter. The T. repens variety ‘Daile’ (form giganteum), one of the often used component of urban greeneries, was studied in this investigation. Plants were grown from seeds and cultivated in the light conditions (18-25 C, 16h/8h of day/night, light intensity 3000 lx) in plastic pots (200 ml), filled with commercial neutralized (pH 5.9 ± 0.3) peat substrate with mineral nutrients. To analyse the impact of increased soil salinity treatments with gradually rising NaCl (0; 20; 40; 60; 80; 100 mM) levels were arranged. Plants were watered when necessary with deionised water to provide optimum substrate moisture 60-70%. The experiment was terminated six weeks after establishment. For analysis of mineral nutrients, dry plant material (above ground part and roots) was used. Decrease of Na content can be significant under elevated salinity till 20 mM NaCl. High NaCl concentrations in the substrate increase Na, Cl, Cu, Fe, and Mn accumulation, but reduce S, Mg, K content in the plant above ground parts. Abiotic stresses generally changes the levels of DNA metilation. Several candidate gene for salt tolerance will be analysed for DNA metilation level using Pyromark-Q24 advanced.

Keywords: DNA metilation, IRAP, soil salinization, white clover

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Authors: James Gallagher, Phillip Benachour

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As location aware mobile technologies become ever more omnipresent, the prospect of exploiting their context awareness to enforce learning approaches thrives. Utilizing the growing acceptance of ubiquitous computing, and the steady progress both in accuracy and battery usage of pervasive devices, we present a case study of remote location viewing, how the application can be utilized to support mobile learning in situ using an existing scenario. Through the case study we introduce a new innovative application: Mobipeek based around a request/response protocol for the viewing of a remote location and explore how this can apply both as part of a teacher lead activity and informal learning situations. The system developed allows a user to select a point on a map, and send a request. Users can attach messages alongside time and distance constraints. Users within the bounds of the request can respond with an image, and accompanying message, providing context to the response. This application can be used alongside a structured learning activity such as the use of mobile phone cameras outdoors as part of an interactive lesson. An example of a learning activity would be to collect photos in the wild about plants, vegetation, and foliage as part of a geography or environmental science lesson. Another example could be to take photos of architectural buildings and monuments as part of an architecture course. These images can be uploaded then displayed back in the classroom for students to share their experiences and compare their findings with their peers. This can help to fosters students’ active participation while helping students to understand lessons in a more interesting and effective way. Mobipeek could augment the student learning experience by providing further interaction with other peers in a remote location. The activity can be part of a wider study between schools in different areas of the country enabling the sharing and interaction between more participants. Remote location viewing can be used to access images in a specific location. The choice of location will depend on the activity and lesson. For example architectural buildings of a specific period can be shared between two or more cities. The augmentation of the learning experience can be manifested in the different contextual and cultural influences as well as the sharing of images from different locations. In addition to the implementation of Mobipeek, we strive to analyse this application, and a subset of other possible and further solutions targeted towards making learning more engaging. Consideration is given to the benefits of such a system, privacy concerns, and feasibility of widespread usage. We also propose elements of “gamification”, in an attempt to further the engagement derived from such a tool and encourage usage. We conclude by identifying limitations, both from a technical, and a mobile learning perspective.

Keywords: context aware, location aware, mobile learning, remote viewing

Procedia PDF Downloads 291

Authors: Ananya Pal, Neelakshi Sarkar, Debraj Saha, Dipanwita Das, Subhashish Kamal Guha, Bibhuti Saha, Runu Chakravarty

Abstract:

Presently, tenofovir disoproxil fumurate (TDF) is the most effective anti-viral agent for the treatment of hepatitis B virus (HBV) in individuals co-infected with HIV and HBV as TDF has activity to suppress both wild-type and lamivudine (3TC)-resistant HBV. However, suboptimal response to TDF was reported in HIV-HBV co-infected individuals with prior 3TC therapy from different countries recently. The incidence of 3TC-resistant HBV strains is quite high in HIV-HBV co-infected patients experiencing long-term anti-retroviral therapy (ART) in eastern India. In spite of this risk, most of the patients with long-term 3TC treatment are continued with the same anti-viral agent in this country. Only a few have received TDF in addition to 3TC in the ART regimen since TDF has been available in India for the treatment of HIV-infected patients in 2012. In this preliminary study, we investigated the virologic and biochemical parameters among HIV-HBV co-infected patients who are non-responders to 3TC treatment during the continuation of 3TC or TDF add-on to 3TC in their ART regimen. Fifteen HIV-HBV co-infected patients who experienced long-term 3TC (mean duration months 36.87 ± 24.08 months) were identified with high HBV viremia ( > 20,000 IU/ml) or harbouring 3TC-resistant HBV. These patients receiving ART from School of Tropical Medicine Kolkata, the main ART centre in eastern India were followed-up semi-annually for next three visits. Different virologic parameters including quantification of plasma HBV load by real-time PCR, detection of hepatitis B e antigen (HBeAg) by commercial ELISA and anti-viral resistant mutations by sequencing were studied. During three follow-up among study subjects, 86%, 47%, and 43% had 3TC-mono-therapy (mean treatment-duration 41.54±18.84, 49.67±11.67, 54.17±12.37 months respectively) whereas 14%, 53%, and 57% experienced TDF in addition to 3TC (mean treatment duration 4.5±2.12, 16.56±11.06, and 23±4.07 months respectively). Mean CD4 cell-count in patients receiving 3TC was tended to be lower during third follow-up as compared to the first and the second [520.67±380.30 (1st), 454.8±196.90 (2nd), and 397.5±189.24 (3rd) cells/mm3) and similar trend was seen in patients experiencing TDF in addition to 3TC [334.5±330.218 (1st), 476.5±194.25 (2nd), and 461.17±269.89 (3rd) cells/mm3]. Serum HBV load was increased during successive follow-up of patients with 3TC-mono-therapy. Initiation of TDF lowered serum HBV-load among 3TC-non-responders at the time of second visit ( < 2,000 IU/ml), interestingly during third follow-up, mean HBV viremia increased >1 log IU/ml (mean 3.56±2.84 log IU/ml). Persistence of 3TC-resistant double and triple mutations was also observed in both the treatment regimens. Mean serum alanine aminotransferase remained elevated in these patients during this follow-up study. Persistence of high HBV viraemia and 3TC-resistant mutation in HBV during the continuation of 3TC might lead to major public health threat in India. The inclusion of TDF in the ART regimen of 3TC non-responder HIV-HBV co-infected patients showed adverse treatment response in terms of virologic and biochemical parameters. Therefore, serious attention is necessary for proper management of long-term 3TC experienced HIV-HBV co-infected patients with high HBV viraemia or 3TC-resistant HBV mutants in India.

Keywords: HBV, HIV, TDF, 3TC-resistant

Procedia PDF Downloads 374

Authors: Alan N. A. Heberle, Salatiel W. Da Silva, Valentin Perez-Herranz, Andrea M. Bernardes

Abstract:

Contaminants of emerging concern are substances widely used, such as pharmaceutical products. These compounds represent risk for both wild and human life since they are not completely removed from wastewater by conventional wastewater treatment plants. In the environment, they can be harm even in low concentration (µ or ng/L), causing bacterial resistance, endocrine disruption, cancer, among other harmful effects. One of the most common taken medicine to treat cardiocirculatory diseases is the Atenolol (ATL), a β-Blocker, which is toxic to aquatic life. In this way, it is necessary to implement a methodology, which is capable to promote the degradation of the ATL, to avoid the environmental detriment. A very promising technology is the advanced electrochemical oxidation (AEO), which mechanisms are based on the electrogeneration of reactive radicals (mediated oxidation) and/or on the direct substance discharge by electron transfer from contaminant to electrode surface (direct oxidation). The hydroxyl (HO•) and sulfate (SO₄•⁻) radicals can be generated, depending on the reactional medium. Besides that, at some condition, the peroxydisulfate (S₂O₈²⁻) ion is also generated from the SO₄• reaction in pairs. Both radicals, ion, and the direct contaminant discharge can break down the molecule, resulting in the degradation and/or mineralization. However, ATL molecule and byproducts can still remain in the treated solution. On this wise, some efforts can be done to implement the AEO process, being one of them the use of a cationic membrane to separate the cathodic (reduction) from the anodic (oxidation) reactor compartment. The aim of this study is investigate the influence of the implementation of a cationic membrane (Nafion®-117) to separate both cathodic and anodic, AEO reactor compartments. The studied reactor was a filter-press, with bath recirculation mode, flow 60 L/h. The anode was an Nb/BDD2500 and the cathode a stainless steel, both bidimensional, geometric surface area 100 cm². The solution feeding the anodic compartment was prepared with ATL 100 mg/L using Na₂SO₄ 4 g/L as support electrolyte. In the cathodic compartment, it was used a solution containing Na₂SO₄ 71 g/L. Between both solutions was placed the membrane. The applied currents densities (iₐₚₚ) of 5, 20 and 40 mA/cm² were studied over 240 minutes treatment time. Besides that, the ATL decay was analyzed by ultraviolet spectroscopy (UV/Vis). The mineralization was determined performing total organic carbon (TOC) in TOC-L CPH Shimadzu. In the cases without membrane, the iₐₚₚ 5, 20 and 40 mA/cm² resulted in 55, 87 and 98 % ATL degradation at the end of treatment time, respectively. However, with membrane, the degradation, for the same iₐₚₚ, was 90, 100 and 100 %, spending 240, 120, 40 min for the maximum degradation, respectively. The mineralization, without membrane, for the same studied iₐₚₚ, was 40, 55 and 72 %, respectively at 240 min, but with membrane, all tested iₐₚₚ reached 80 % of mineralization, differing only in the time spent, 240, 150 and 120 min, for the maximum mineralization, respectively. The membrane increased the ATL oxidation, probably due to avoid oxidant ions (S₂O₈²⁻) reduction on the cathode surface.

Keywords: contaminants of emerging concern, advanced electrochemical oxidation, atenolol, cationic membrane, double compartment reactor

Procedia PDF Downloads 136

Authors: Ocsana Opris, Ildiko Lung, Maria L. Soran, Alexandra Ciorita, Lucian Copolovici

Abstract:

The effects assessment of environmental stress factors on both crop and wild plants of nutritional value are a very important research topic. Continuously worldwide consumption of drugs leads to significant environmental pollution, thus generating environmental stress. Understanding the effects of the important drugs on plant composition and ultrastructural modification is still limited, especially at environmentally relevant concentrations. The aim of the present work was to investigate the influence of three non-steroidal anti-inflammatory drugs (NSAIDs) on chlorophylls content, carotenoids content, total polyphenols content, antioxidant capacity, and ultrastructure of orache (Atriplex patula L.) and spinach (Spinacia oleracea L.). All green leafy vegetables selected for this study were grown in controlled conditions and treated with solutions of different concentrations (0.1‒1 mg L⁻¹) of diclofenac, ibuprofen, and naproxen. After eight weeks of exposure of the plants to NSAIDs, the chlorophylls and carotenoids content were analyzed by high-performance liquid chromatography coupled with photodiode array and mass spectrometer detectors, total polyphenols and antioxidant capacity by ultraviolet-visible spectroscopy. Also, the ultrastructural analyses of the vegetables were performed using transmission electron microscopy in order to assess the influence of the selected NSAIDs on cellular organisms, mainly photosynthetic organisms (chloroplasts), energy supply organisms (mitochondria) and nucleus as a cellular metabolism coordinator. In comparison with the control plants, decreases in the content of chlorophylls were observed in the case of the Atriplex patula L. plants treated with ibuprofen (11-34%) and naproxen (25-52%). Also, the chlorophylls content from Spinacia oleracea L. was affected, the lowest decrease (34%) being obtained in the case of the treatment with naproxen (1 mg L⁻¹). Diclofenac (1 mg L⁻¹) affected the total polyphenols content (a decrease of 45%) of Atriplex patula L. and ibuprofen (1 mg L⁻¹) affected the total polyphenols content (a decrease of 20%) of Spinacia oleracea L. The results obtained also indicate a moderate reduction of carotenoids and antioxidant capacity in the treated plants, in comparison with the controls. The investigations by transmission electron microscopy demonstrated that the green leafy vegetables were affected by the selected NSAIDs. Thus, this research contributes to a better understanding of the adverse effects of these drugs on studied plants. Important to mention is that the dietary intake of these drugs contaminated plants, plants with important nutritional value, may also presume a risk to human health, but currently little is known about the fate of the drugs in plants and their effect on or risk to the ecosystem.

Keywords: abiotic stress, green leafy vegetables, pigments content, ultra structure

Procedia PDF Downloads 125

Authors: Alaa G. M. Osman, Abd-El –Baset M. Abd El Reheem, Khaled Y. Abouelfadl, Usama M. Mahmoud, Mohsen A. Moustafa

Abstract:

This study aimed to explore new sites of biomarker research and to establish the use of blood parameters in wild fish populations. Four hundred and twenty fish samples were collected from six sites along the whole course of the river Nile, Egypt. The mean values of erythrocytes, thrombocytes, hemoglobin concentration, hematocrit value, and mean corpuscular volume were significantly lower in the blood of Nile tilapia and African catfish collected from downstream (contaminated) compared to upstream sites. In contrast, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in the peripheral blood of both fish species significantly increased from upstream to downstream river Nile. The leukocytes count was significantly decreased in contaminated sites compared to upstream area. Hematological variables in the peripheral blood of Oreochromis niloticus niloticus and Clarias gariepinus exhibited significant (p<0.05) correlation with nearly all the detected chemical and physical parameters along the Nile course. In the present study, lower cellular and nuclear areas and cellular and nuclear shape factor were recorded in the erythrocytes of fish collected from downstream compared to those caught from upstream sites. This was confirmed by higher immature ratios of red cells in the blood of fish sampled from downstream river Nile. Karyorrhetic and enucleated erythrocytes were significantly correlated with physiochemical parameters in water samples collected from the same sites is being higher in the blood of fish collected from downstream sites. To see if there was any correlation between fish altered physiological fitness and environmental stress, we measured serum biochemical variables namely; total protein, cholesterol, triglycerides, calcium, chlorides, alkaline phosphatase activity (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid activity, creatinine, and serum glucose. The level of all the selected biochemical variables in the blood of O. niloticus niloticus and C. gariepinus were recorded to be significantly higher (p<0.05) in downstream sites. According to the present results, nearly all the detected haematological and blood biochemical variables are suitable indicators of contaminant exposure in O. niloticus niloticus and C. gariepinus. Also the detected erythrocytes malformations in blood collected from Nile tilapia and African catfish were proven to be suitable for bio-monitoring aquatic pollution. The results revealed species-specific differences in sensitivities, suggesting that Nile tilapia may serve as a more sensitive test species compared to African catfish.

Keywords: biomarkers, water pollution, blood parameters, river nile, african catfish, nile tilapia

Procedia PDF Downloads 291

Authors: Baye Berihun Getahun, Mulugeta Atnaf Tiruneh, Richard G. F. Visser

Abstract:

Resource-poor farmers practice low-input farming systems, and yet, most breeding programs give less attention to this huge farming system, which serves as a source of food and income for several people in developing countries. The high-input conventional breeding system appears to have failed to adequately meet the needs and requirements of 'difficult' environments operating under this system. Moreover, the unavailability of resources for crop production is getting for their peaks, the environment is maltreated by excessive use of agrochemicals, crop productivity reaches its plateau stage, particularly in the developed nations, the world population is increasing, and food shortage sustained to persist for poor societies. In various parts of the world, genetic gain at the farmers' level remains low which could be associated with low adoption of crop varieties, which have been developed under high input systems. Farmers usually use their local varieties and apply minimum inputs as a risk-avoiding and cost-minimizing strategy. This evidence indicates that the conventional high-input plant breeding system has failed to feed the world population, and the world is moving further away from the United Nations' goals of ending hunger, food insecurity, and malnutrition. In this review, we discussed the rationality of focused breeding programs for low-input farming systems and, the technical aspect of crop breeding that accommodates future food needs and its significance for developing countries in the decreasing scenario of resources required for crop production. To this end, the application of exotic introgression techniques like polyploidization, pan-genomics, comparative genomics, and De novo domestication as a pre-breeding technique has been discussed in the review to exploit the untapped genetic diversity of the crop wild relatives (CWRs). Desired recombinants developed at the pre-breeding stage are exploited through appropriate breeding approaches such as evolutionary plant breeding (EPB), rhizosphere-related traits breeding, and participatory plant breeding approaches. Populations advanced through evolutionary breeding like composite cross populations (CCPs) and rhizosphere-associated traits breeding approach that provides opportunities for improving abiotic and biotic soil stress, nutrient acquisition capacity, and crop microbe interaction in improved varieties have been reviewed. Overall, we conclude that low input farming system is a huge farming system that requires distinctive breeding approaches, and the exotic pre-breeding introgression techniques and the appropriate breeding approaches which deploy the skills and knowledge of both breeders and farmers are vital to develop heterogeneous landrace populations, which are effective for farmers practicing low input farming across the world.

Keywords: low input farming, evolutionary plant breeding, composite cross population, participatory plant breeding

Procedia PDF Downloads 50

Authors: Anisuzzaman Khan, A. J. K. Masud

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Bangladesh is the largest delta in the world predominantly comprising large network of rives and wetlands. Wetlands in Bangladesh are represented by inland freshwater, estuarine brakishwater and tidal salt-water coastal wetlands. Bangladesh possesses enormous area of wetlands including rivers and streams, freshwater lakes and marshes, haors, baors, beels, water storage reservoirs, fish ponds, flooded cultivated fields and estuarine systems with extensive mangrove swamps. The past, present, and future of Bangladesh, and its people’s livelihoods are intimately connected to its relationship with water and wetlands. More than 90% of the country’s total area consists of alluvial plains, crisscrossed by a complex network of rivers and their tributaries. Floodplains, beels (low-lying depressions in the floodplain), haors (deep depression) and baors (oxbow lakes) represent the inland freshwater wetlands. Over a third of Bangladesh could be termed as wetlands, considering rivers, estuaries, mangroves, floodplains, beels, baors and haors. The country’s wetland ecosystems also offer critical habitats for globally significant biological diversity. Of these the deeply flooded basins of north-east Bangladesh, known as haors, are a habitat of wide range of wild flora and fauna unique to Bangladesh. The haor basin lies within the districts of Sylhet, Sunamgonj, Netrokona, Kishoregonj, Habigonj, Moulvibazar, and Brahmanbaria in the Northeast region of Bangladesh comprises the floodplains of the Meghna tributaries and is characterized by the presence of numerous large, deeply flooded depressions, known as haors. It covers about around 8,568 km2 area of Bangladesh. The topography of the region is steep at around foothills in the north and slopes becoming mild and milder gradually at downstream towards south. Haor is a great reservoir of aquatic biological resources and acts as the ecological safety net to the nature as well as to the dwellers of the haor. But in reality, these areas are considered as wastelands and to make these wastelands into a productive one, a one sided plan has been implementing since long. The programme is popularly known as Flood Control, Drainage and Irrigation (FCDI) which is mainly devoted to increase the monoculture rice production. However, haor ecosystem is a multiple-resource base which demands an integrated sustainable development approach. The ongoing management approach is biased to only rice production through FCDI. Thus this primitive mode of action is diminishing other resources having more economic potential ever thought.

Keywords: freshwater wetlands, biological diversity, biological resources, conservation and sustainable development

Procedia PDF Downloads 329

Authors: Apurva Singh, S. P. Jaiswar, Apala Priyadarshini, Akancha Pandey

Abstract:

Objective: The aim of this study is to find the association of PON1 gene polymorphism with pesticides In RPL subjects. Background: Recurrent pregnancy loss (RPL) is defined as three or more sequential abortions before the 20th week of gestation. Pesticides and its derivatives (organochlorine and organophosphate) are proposed to accommodate a ruler chemical for RPL in the sub-humid region of India. The paraoxonase-1 enzyme (PON1) plays an important role in the toxicity of some organophosphate pesticides, with low PON1 activity being associated with higher pesticide sensitivity Methodology: This is a case-control study done in Department of Obstetrics & Gynaecology & Department of Biochemistry, K.G.M.U, Lucknow, India. The subjects were enrolled after fulfilling the inclusion & exclusion criteria. Inclusion criteria: Cases- Subject having two or more spontaneous abortions & Control- Healthy female having one or more alive child was selected. Exclusion criteria: Cases & Control- Subject having the following disease will be excluded from the study Diabetes mellitus, Hypertension, Tuberculosis, Immunocompromised patients, any endocrine disorder and genital, colon or breast cancer any other malignancies. Blood samples were collected in EDTA tubes from cases & healthy control women & genomic DNA was extracted by phenol-chloroform method. The estimation of pesticides residue from blood was done by HPLC. Biochemical estimation was also performed. Genotyping of PON1 gene polymorphism was performed by RFLP. Statistical analysis of the data was performed using the SPSS16.3 software. Results: A sum of total 14 pesticides (12 organochlorine and 2 organophosphate) selected on the basis of their persistent nature and consumption rate. The significant level of pesticide (ppb) estimated by the Mann whiney test and it was found to be significant at higher level of β-HCH (p:0.04), γ-HCH (p:0.001), δ-HCH (p: 0.002), chloropyrifos (p:0.001), pp-DDD (p:0.001) and fenvalrate (p: 0.001) in case group compare to its control. The level of antioxidant enzymes were found to be significantly decreased among the cases. Wild homozygous TT was more frequent and prevalent among control groups. However, heterozygous group (Tt) was more in cases than control groups (CI-0.3-1.3) (p=0.06). Conclusion: Higher levels of pesticides with endocrine disrupting potential in cases indicate the possible role of these compounds as one of the causes of recurrent pregnancy loss. Possibly, increased pesticide level appears to indicate increased levels of oxidative damage that has been associated with the possible cause of Recurrent Miscarriage, it may reflect indirect evidence of toxicity rather than the direct cause. Since both factors are reported to increase risk, individuals with higher levels of these 'Toxic compounds' especially in 'high-risk genotypes' might be more susceptible to recurrent pregnancy loss.

Keywords: paraoxonase, pesticides, PON1, RPL

Procedia PDF Downloads 143

Authors: Thaís T. Valério Caetano, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

Abstract:

Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil for a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: nicotiana glauca graham, global invasive species database, alkaloids, toxic

Procedia PDF Downloads 88

Authors: Amanda Gregorim Fernandes, Lorena Cardoso Cintra, Rosalia Santos Amorim Jesuino, Fabricia Paula De Faria, Marcio José Poças Fonseca

Abstract:

Bioethanol is one of the most promising biofuels and able to replace fossil fuels and reduce its different environmental impacts and can be generated from various agroindustrial waste. The Brazil is in first place in bioethanol production to be the largest producer of sugarcane. The bagasse sugarcane (SCB) has lignocellulose which is composed of three major components: cellulose, hemicellulose and lignin. Cellulose is a homopolymer of glucose units connected by glycosidic linkages. Among all species of Penicillium, Penicillium echinulatum has been the focus of attention because they produce high quantities of cellulase and the mutant strain 9A02S1 produces higher enzyme levels compared to the wild. Among the cellulases, the cellobiohydrolases enzymes are the main components of the cellulolytic system of fungi, and are also responsible for most of the potential hydrolytic in enzyme cocktails for the industrial processing of plant biomass and several cellobiohydrolases Penicillium had higher specific activity against cellulose compared to CBH I from Trichoderma reesei. This fact makes it an interesting pattern for higher yields in the enzymatic hydrolysis, and also they are important enzymes in the hydrolysis of crystalline regions of cellulose. Therefore, finding new and more active enzymes become necessary. Meanwhile, β-glycosidases act on soluble substrates and are highly dependent on cellobiohydrolases and endoglucanases action to provide the substrate in the hydrolysis of the biomass, but the cellobiohydrolases and endoglucanases are highly dependent β-glucosidases to maintain efficient hydrolysis. Thus, there is a need to understand the structure-function relationships that govern the catalytic activity of cellulolytic enzymes to elucidate its mechanism of action and optimize its potential as industrial biocatalysts. To evaluate the enzyme β-glucosidase of Penicillium echinulatum (PeBGL1) the gene was synthesized from the assembly sequence from a library in induction conditions and then the PeBGL1 gene was cloned in the vector pPICZαA and transformed into P. pastoris GS115. After processing, the producers of PeBGL1 were analyzed for enzyme activity and protein profile where a band of approximately 100 kDa was viewed. It was also carried out the zymogram. In partial characterization it was determined optimum temperature of 50°C and optimum pH of 6,5. In addition, to increase the secreted recombinant PeBGL1 production by Pichia pastoris, three parameters of P. pastoris culture medium were analysed: methanol, nitrogen source concentrations and the inoculum size. A 23 factorial design was effective in achieving the optimum condition. Altogether, these results point to the potential application of this P. echinulatum β-glucosidase in hydrolysis of cellulose for the production of bioethanol.

Keywords: bioethanol, biotechnology, beta-glucosidase, penicillium echinulatum

Procedia PDF Downloads 242

Authors: Patrícia Branco, Catarina Prista, Helena Albergaria

Abstract:

Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.

Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts

Procedia PDF Downloads 99

Authors: Thaís T. Valério Caetano, Lívia de Carvalho Ferreira, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

Abstract:

Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil during a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: alkaloid production, invasive species, nicotiana glauca, plant phenology

Procedia PDF Downloads 83

Authors: Sijie Chen, Xiaofang Chen, Juan Wang, Yingying Zhang, Yu Hong, Wanyu Zhuang, Xinxin Huang, Ping Ou, Longsheng Huang

Abstract:

Purpose: Autistic symptom improvement can be observed in children treated with acupuncture, but the mechanism is still being explored. In the present study, we used scalp acupuncture to treat autism rat model, and then their improvement in the abnormal behaviors and specific mechanisms behind were revealed by detecting animal behaviors, analyzing the RNA sequencing of the prefrontal cortex(PFC), and observing the ultrastructure of PFC neurons under the transmission electron microscope. Methods: On gestational day 12.5, Wistar rats were given valproic acid (VPA) by intraperitoneal injection, and their offspring were considered to be reliable rat models of autism. They were randomized to VPA or VPA-acupuncture group (n=8). Offspring of Wistar pregnant rats that were simultaneously injected with saline were randomly selected as the wild-type group (WT). VPA_acupuncture group rats received acupuncture intervention at 23 days of age for 4 weeks, and the other two groups followed without intervention. After the intervention, all experimental rats underwent behavioral tests. Immediately afterward, they were euthanized by cervical dislocation, and their prefrontal cortex was isolated for RNA sequencing and transmission electron microscopy. Results: The main results are as follows: 1. Animal behavioural tests: VPA group rats showed more anxiety-like behaviour and repetitive, stereotyped behaviour than WT group rats. While VPA group rats showed less spatial exploration ability, activity level, social interaction, and social novelty preference than WT group rats. It was gratifying to observe that acupuncture indeed improved these abnormal behaviors of autism rat model. 2. RNA-sequencing: The three groups of rats differed in the expression and enrichment pathways of multiple genes related to synaptic function, neural signal transduction, and circadian rhythm regulation. Our experiments indicated that acupuncture can alleviate the major symptoms of ASD by improving these neurological abnormalities. 3. Under the transmission electron microscopy, several lysosomes and mitochondrial structural abnormalities were observed in the prefrontal neurons of VPA group rats, which were manifested as atrophy of the mitochondrial membran, blurring or disappearance of the mitochondrial cristae, and even vacuolization. Moreover, the number of synapses and synaptic vesicles was relatively small. Conversely, the mitochondrial structure of rats in the WT group and VPA_acupuncture was normal, and the number of synapses and synaptic vesicles was relatively large. Conclusion: Acupuncture effectively improved the abnormal behaviors of autism rat model and the ultrastructure of the PFC neurons, which might worked by improving their abnormal synaptic function, synaptic plasticity and promoting neuronal signal transduction.

Keywords: autism spectrum disorder, acupuncture, animal behavior, RNA sequencing, transmission electron microscope

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Authors: P.P. Rupasinghe, A.H. Farid

Abstract:

The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.

Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution

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Authors: Ioannis Panagiotopoulos, Aristidis Anastasakis, Konstantinos Toutouzas, Ioannis Iakovou, Charalampos Vlachopoulos, Vasilis Voudris, Georgios Tziomalos, Konstantinos Tsioufis, Efstathios Kastritis, Alexandros Briassoulis, Kimon Stamatelopoulos, Alexios Antonopoulos, Paraskevi Exadaktylou, Evanthia Giannoula, Anastasia Katinioti, Maria Kalantzi, Evangelos Leontiadis, Eftychia Smparouni, Ioannis Malakos, Nikolaos Aravanis, Argyrios Doumas, Maria Koutelou

Abstract:

Introduction: Cardiac amyloidosis from Transthyretin (ATTR-CA) is an infiltrative disease characterized by the deposition of pathological transthyretin complexes in the myocardium. This study describes the characteristics of patients diagnosed with ATTR-CA from 2019 until present at the Nuclear Medicine Department of Onassis Cardiac Surgery Center and AHEPA Hospital. These centers have extensive experience in amyloidosis and modern technological equipment for its diagnosis. Materials and Methods: Records of consecutive patients (N=73) diagnosed with any type of amyloidosis were collected, analyzed, and prospectively followed. The diagnosis of amyloidosis was made using specific myocardial scintigraphy with Tc-99m DPD. Demographic characteristics, including age, gender, marital status, height, and weight, were collected in a database. Clinical characteristics, such as amyloidosis type (ATTR and AL), serum biomarkers (BNP, troponin), electrocardiographic findings, ultrasound findings, NYHA class, aortic valve replacement, device implants, and medication history, were also collected. Some of the most significant results are presented. Results: A total of 73 cases (86% male) were diagnosed with amyloidosis over four years. The mean age at diagnosis was 82 years, and the main symptom was dyspnea. Most patients suffered from ATTR-CA (65 vs. 8 with AL). Out of all the ATTR-CA patients, 61 were diagnosed with wild-type and 2 with two rare mutations. Twenty-eight patients had systemic amyloidosis with extracardiac involvement, and 32 patients had a history of bilateral carpal tunnel syndrome. Four patients had already developed polyneuropathy, and the diagnosis was confirmed by DPD scintigraphy, which is known for its high sensitivity. Among patients with isolated cardiac involvement, only 6 had left ventricular ejection fraction below 40%. The majority of ATTR patients underwent tafamidis treatment immediately after diagnosis. Conclusion: In conclusion, the experiences shared by the two centers and the continuous exchange of information provide valuable insights into the diagnosis and management of cardiac amyloidosis. Clinical suspicion of amyloidosis and early diagnostic approach are crucial, given the availability of non-invasive techniques. Cardiac scintigraphy with DPD can confirm the presence of the disease without the need for a biopsy. The ultimate goal still remains continuous education and awareness of clinical cardiologists so that this systemic and treatable disease can be diagnosed and certified promptly and treatment can begin as soon as possible.

Keywords: amyloidosis, diagnosis, myocardial scintigraphy, Tc-99m DPD, transthyretin

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Authors: Paula I. Buonfiglio, Carlos David Bruque, Lucia Salatino, Vanesa Lotersztein, Sebastián Menazzi, Paola Plazas, Ana Belén Elgoyhen, Viviana Dalamón

Abstract:

Hearing loss (HL) is the most prevalent sensorineural disorder affecting about 10% of the global population, with more than half due to genetic causes. About 1 in 500-1000 newborns present congenital HL. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance. To date, more than 100 genes are related to HL. Therefore, the Whole-exome sequencing (WES) technique has become a cost-effective alternative approach for molecular diagnosis. Nevertheless, new challenges arise from the detection of novel variants, in particular missense changes, which can lead to a spectrum of genotype-to-phenotype correlations, which is not always straightforward. In this work, we aimed to identify the genetic causes of HL in isolated and familial cases by designing a multistep approach to analyze target genes related to hearing impairment. Moreover, we performed in silico and in vivo analyses in order to further study the effect of some of the novel variants identified in the hair cell function using the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes, respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Fifty of the undiagnosed patients with moderate HL were tested for deletions in STRC gene by Multiplex ligation-dependent probe amplification technique (MLPA), leading to 6% of diagnosis. After this initial screening, 50 families were selected to be analyzed by WES, achieving diagnosis in 44% of them. Half of the identified variants were novel. A missense variant in MYO6 gene detected in a family with postlingual HL was selected to be further analyzed. A protein modeling with AlphaFold2 software was performed, proving its pathogenic effect. In order to functionally validate this novel variant, a knockdown phenotype rescue assay in zebrafish was carried out. Injection of wild-type MYO6 mRNA in embryos rescued the phenotype, whereas using the mutant MYO6 mRNA (carrying c.2782C>A variant) had no effect. These results strongly suggest the deleterious effect of this variant on the mobility of stereocilia in zebrafish neuromasts, and hence on the auditory system. In the present work, we demonstrated that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. These results highlight the importance of a combined strategy in order to identify candidate variants as well as the in silico and in vivo studies to analyze and prove their pathogenicity and accomplish a better understanding of the mechanisms underlying the physiopathology of the hearing impairment.

Keywords: diagnosis, genetics, hearing loss, in silico analysis, in vivo analysis, WES, zebrafish

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Authors: Luis M. Ribeiro, Jorge Raposo, Ricardo Oliveira, David Caballero, Domingos X. Viegas

Abstract:

Under the scope of European project WUIWATCH a set of experimental tests on house vulnerability was performed in order to assess the resistance of selected house components during the passage of a forest fire. Among the individual elements most affected by the passage of a wildfire the windows are the ones with greater exposure. In this sense, a set of exploratory experimental tests was designed to assess some particular aspects related to the vulnerability of windows and blinds. At the same time, the importance of leaving them closed (as well as the doors inside a house) during a wild fire was explored in order to give some scientific background to guidelines for homeowners. Three sets of tests were performed: 1. Windows and blinds resistance to heat. Three types of protective blinds were tested (aluminium, PVC and wood) on 2 types of windows (single and double pane). The objective was to assess the structures resistance. 2. The influence of air flow on the transport of burning embers inside a house. A room was built to scale, and placed inside a wind tunnel, with one window and one door on opposite sides. The objective was to assess the importance of leaving an inside door opened on the probability of burning embers entering the room. 3. The influence of the dimension of openings on a window or door related to the probability of ignition inside a house. The objective was to assess the influence of different window openings in relation to the amount of burning particles that can enter a house. The main results were: 1. The purely radiative heat source provides 1.5 KW/m2 of heat impact in the structure, while the real fire generates 10 Kw/m2. When protected by the blind, the single pane window reaches 30ºC on both sides, and the double pane window has a differential of 10º from the side facing the heat (30ºC) and the opposite side (40ºC). Unprotected window constantly increases temperature until the end of the test. Window blinds reach considerably higher temperatures. PVC loses its consistency above 150ºC and melts. 2. Leaving the inside door closed results in a positive pressure differential of +1Pa from the outside to the inside, inhibiting the air flow. Opening the door in half or full reverts the pressure differential to -6 and -8 times respectively, favouring the air flow from the outside to the inside. The number of particles entering the house follows the same tendency. 3. As the bottom opening in a window increases from 0,5 cm to 4 cm the number of particles that enter the house per second also increases greatly. From 5 cm until 80cm there is no substantial increase in the number of entering particles. This set of exploratory tests proved to be an added value in supporting guidelines for home owners, regarding self-protection in WUI areas.

Keywords: forest fire, wildland urban interface, house vulnerability, house protective elements

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