Search results for: species profile

Authors: Sudhanshu Sharma

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Lisinopril, 1-[N-{(s)-I-carboxy-3 phenyl propyl}-L-proline dehydrate is a lysine analog of enalaprilat, the active metabolite of enalapril. It is long-acting, non-sulhydryl angiotensin-converting enzyme (ACE) inhibitor that is used for the treatment of hypertension and congestive heart failure in daily dosage 10-80 mg. Pharmacological activity of lisinopril has been proved in various experimental and clinical studies. Owing to its importance and widespread use, efforts have been made towards the development of simple and reliable analytical methods. As per our literature survey, lisinopril in pharmaceutical formulations has been determined by various analytical methodologies like polaragraphy, potentiometry, and spectrophotometry, but most of these analytical methods are not too suitable for the Identification of lisinopril from clinical samples because of the interferences caused by the amino acids and amino groups containing metabolites present in biological samples. This report is an attempt in the direction of developing a simple and reliable method for on plate identification and quantification of lisinopril in pharmaceutical formulations as well as from human urine samples using silica gel H layers developed with a new mobile phase comprising of micellar solutions of N-cetyl-N, N, N-trimethylammonium bromide (CTAB). Micellar solutions have found numerous practical applications in many areas of separation science. Micellar liquid chromatography (MLC) has gained immense popularity and wider applicability due to operational simplicity, cost effectiveness, relatively non-toxicity and enhanced separation efficiency, low aggressiveness. Incorporation of aqueous micellar solutions as mobile phase was pioneered by Armstrong and Terrill as they accentuated the importance of TLC where simultaneous separation of ionic or non-ionic species in a variety of matrices is required. A peculiarity of the micellar mobile phases (MMPs) is that they have no macroscopic analogues, as a result the typical separations can be easily achieved by using MMPs than aqueous organic mobile phases. Previously MMPs were successfully employed in TLC based critical separations of aromatic hydrocarbons, nucleotides, vitamin K1 and K5, o-, m- and p- aminophenol, amino acids, separation of penicillins. The human urine analysis for identification of selected drugs and their metabolites has emerged as an important investigation tool in forensic drug analysis. Among all chromatographic methods available only thin layer chromatography (TLC) enables a simple fast and effective separation of the complex mixtures present in various biological samples and is recommended as an approved testing for forensic drug analysis by federal Law. TLC proved its applicability during successful separation of bio-active amines, carbohydrates, enzymes, porphyrins, and their precursors, alkaloid and drugs from urine samples.

Keywords: lisnopril, surfactant, chromatography, micellar solutions

Procedia PDF Downloads 341

Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

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Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

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Authors: Yanto Santosa, Dede Aulia Rahman, Cory Wulan, Abdul Haris Mustari

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The global demand for long-tailed macaques for medical experimentation has continued to increase. Fulfillment of Indonesian export demands has been mostly from natural habitats, based on a harvesting quota. This quota has been determined according to the total catch for a given year, and not based on consideration of any demographic parameters or physical environmental factors with regard to the animal; hence threatening the sustainability of the various populations. It is therefore necessary to formulate a method for calculating a sustainable harvesting quota, based on population parameters in natural habitats. Considering the possibility of variations in habitat characteristics and population parameters, a time series observation of demographic and physical/biotic parameters, in various habitats, was performed on 13 groups of long-tailed macaques, distributed throughout the West Java, Lampung and Yogyakarta areas of Indonesia. These provinces were selected for comparison of the influence of human/tourism activities. Data on population parameters that was collected included data on life expectancy according to age class, numbers of individuals by sex and age class, and ‘ratio of infants to reproductive females’. The estimation of population growth was based on a population dynamic growth model: the Leslie matrix. The harvesting quota was calculated as being the difference between the actual population size and the MVP (minimum viable population) for each sex and age class. Observation indicated that there were variations within group size (24 – 106 individuals), gender (sex) ratio (1:1 to 1:1.3), life expectancy value (0.30 to 0.93), and ‘ratio of infants to reproductive females’ (0.23 to 1.56). Results of subsequent calculations showed that sustainable harvesting quotas for each studied group of long-tailed macaques, ranged from 29 to 110 individuals. An estimation model of the MVP for each age class was formulated as Log Y = 0.315 + 0.884 Log Ni (number of individual on ith age class). This study also found that life expectancy for the juvenile age class was affected by the humidity under tree stands, and dietary plants’ density at sapling, pole and tree stages (equation: Y= 2.296 – 1.535 RH + 0.002 Kpcg – 0.002 Ktg – 0.001 Kphn, R2 = 89.6% with a significance value of 0.001). By contrast, for the sub-adult-adult age class, life expectancy was significantly affected by slope (equation: Y=0.377 = 0.012 Kml, R2 = 50.4%, with significance level of 0.007). The infant to reproductive female ratio was affected by humidity under tree stands, and dietary plant density at sapling and pole stages (equation: Y = -1.432 + 2.172 RH – 0.004 Kpcg + 0.003 Ktg, R2 = 82.0% with significance level of 0.001). This research confirmed the importance of population parameters in determining the minimum viable population, and that MVP varied according to habitat characteristics (especially food availability). It would be difficult therefore, to formulate a general mathematical equation model for determining a harvesting quota for the species as a whole.

Keywords: harvesting, long-tailed macaque, population, quota

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Authors: A. J. Olatunji, Adebolu Temitope Johnson

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Groundwater samples were collected randomly from hand-dug wells and boreholes in parts of the Ado Ekiti metropolis and were subjected to quality assessment and characterization. Physicochemical analyses, which include the in-situ parameters (pH units, Turbidity, and Electrical Conductivity) and laboratory analysis of selected ionic concentrations, were carried out following standard methods. Hydrochemistry of the present study revealed relative mean concentrations of cations in the order Ca2+ > Na+ > Mg2+ > Cu2+> Fe > Mn2+ and that of anions: Cl- > NO3- > SO42- > F - respectively considering World Health Organisation Standard (WHO) range of values for potable water. The result shows that values of certain parameters (Total Dissolved Solid (TDS), Manganese, Calcium, Magnesium, Fluoride, and Sulphate) were below the Highest Desirable Level of the Standards, while values of some other parameters (pH Units, Electrical Conductivity, Turbidity, Alkalinity, Sodium, Copper, Chloride, and Total Hardness) were within the range of figures between Highest Desirable Level (HDL) and Maximum Permissible Level (MPL) of World Health Organization (WHO) drinking water Standards. The reduction in the mean concentration value of Total Dissolved Solids (TDS) of most borehole samples follows the fact that water had been allowed to settle in the overhead tanks before usage; we discussed and brainstormed in the course of sampling and agreed to take a sample that way because that represents what the people consume, it also shows an indication while there was slightly concentration increase of these soluble ions in hand-dug wells samples than borehole samples only with the exception of borehole sample seven BH7 because BH7 uses the mono-pumping system. These in-situ parameters and ionic concentrations were further displayed and or represented on bar charts along with the WHO standards for better pictorial clarifications. Deductions from field observation indices revealed the imprints of natural weathering, ion-exchange processes, and anthropogenic activities influencing groundwater quality. A strong degree of association was found to exist between sodium and chlorine ions in both hand-dug well and borehole groundwater samples through the use of Pearson’s correlation coefficient; this association can further be supported by the chemistry of the parent bedrock associated with the study area because the chemistry of groundwater is a replica of its host rock. The correlation of those two ions must have begun from the period of mountain building, indicating an identical source from which they were released to the groundwater. Moreover, considering the comparison of ionic species concentrations of all samples with the (WHO) standards, there were no anomalous increases or decreases in the laboratory analysis results; this simply reveals an insignificant state of pollution of the groundwater. The study and its sampling techniques were not set to target the likely area and extent of groundwater pollution but its portability. It could be said that the samples were safe for human consumption.

Keywords: groundwater, physicochemical, parameters ionic, concentrations, WHO standards

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Authors: Majid Hejazian, Nam-Trung Nguyen

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Over the past few years, microfluidic devices have generated significant attention from industry and academia due to advantages such as small sample volume, low cost and high efficiency. Microfluidic devices have applications in chemical, biological and industry analysis and can facilitate assay of bio-materials and chemical reactions, separation, and sensing. Micromixers are one of the important microfluidic concepts. Micromixers can work as stand-alone devices or be integrated in a more complex microfluidic system such as a lab on a chip (LOC). Micromixers are categorized as passive and active types. Passive micromixers rely only on the arrangement of the phases to be mixed and contain no moving parts and require no energy. Active micromixers require external fields such as pressure, temperature, electric and acoustic fields. Rapid and efficient mixing is important for many applications such as biological, chemical and biochemical analysis. Achieving fast and homogenous mixing of multiple samples in the microfluidic devices has been studied and discussed in the literature recently. Improvement in mixing rely on effective mass transport in microscale, but are currently limited to molecular diffusion due to the predominant laminar flow in this size scale. Using magnetic field to elevate mass transport is an effective solution for mixing enhancement in microfluidics. The use of a non-uniform magnetic field to improve mass transfer performance in a microfluidic device is demonstrated in this work. The phenomenon of mixing ferrofluid and DI-water streams has been reported before, but mass transfer enhancement for other non-magnetic species through magnetic field have not been studied and evaluated extensively. In the present work, permanent magnets were used in a simple microfluidic device to create a non-uniform magnetic field. Two streams are introduced into the microchannel: one contains fluorescent dye mixed with diluted ferrofluid to induce enhanced mass transport of the dye, and the other one is a non-magnetic DI-water stream. Mass transport enhancement of fluorescent dye is evaluated using fluorescent measurement techniques. The concentration field is measured for different flow rates. Due to effect of magnetic field, a body force is exerted on the paramagnetic stream and expands the ferrofluid stream into non-magnetic DI-water flow. The experimental results demonstrate that without a magnetic field, both magnetic nanoparticles of the ferrofluid and the fluorescent dye solely rely on molecular diffusion to spread. The non-uniform magnetic field, created by the permanent magnets around the microchannel, and diluted ferrofluid can improve mass transport of non-magnetic solutes in a microfluidic device. The susceptibility mismatch between the fluids results in a magnetoconvective secondary flow towards the magnets and subsequently the mass transport of the non-magnetic fluorescent dye. A significant enhancement in mass transport of the fluorescent dye was observed. The platform presented here could be used as a microfluidics-based micromixer for chemical and biological applications.

Keywords: ferrofluid, mass transfer, micromixer, microfluidics, magnetic

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Authors: Carolina Gheller Miguens

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Feeding is, and should be, pleasurable for living beings so they desire to nourish themselves while preserving the continuity of the species. Social rites usually revolve around the table and are closely linked to the cultural traditions of each region and social group. Since the beginning, food has been closely linked with the products each region provides, and, also, related to the respective seasons of production. With the globalization and facilities of modern life we are able to find an ever increasing variety of products at any time of the year on supermarket shelves. These lifestyle changes end up directly influencing food traditions. With the era of uncontrolled obesity caused by the dazzle with the large and varied supply of low-priced to ultra-processed industrial products now in the past, today we are living a time when people are putting aside the pleasure of eating to exclusively eat food dictated by the media as healthy. Recently the medicalization of food in our society has become so present in daily life that almost without realizing we make food choices conditioned to the studies of the properties of these foods. The fact that people are more attentive to their health is interesting. However, when this care becomes an obsessive disorder, which imposes itself on the pleasure of eating and extinguishes traditional customs, it becomes dangerous for our recognition as citizens belonging to a culture and society. This new way of living generates a rupture with the social environment of origin, possibly exposing old traditions to oblivion after two or three generations. Based on these facts, the presented study analyzes these social transformations that occur in our society that triggered the current medicalization of food. In order to clarify what is actually a healthy diet, this research proposes a critical analysis on the subject aiming to understand nutritional rationality and relate how it acts in the medicalization of food. A wide bibliographic review on the subject was carried out followed by an exploratory research in online (especially social) media, a relevant source in this context due to the perceived influence of such media in contemporary eating habits. Finally, this data was crossed, critically analyzing the current situation of the concept of healthy eating and medicalization of food. Throughout this research, it was noticed that people are increasingly seeking information about the nutritional properties of food, but instead of seeking the benefits of products that traditionally eat in their social environment, they incorporate external elements that often bring benefits similar to the food already consumed. This is because the access to information is directed by the media and exalts the exotic, since this arouses more interest of the population in general. Efforts must be made to clarify that traditional products are also healthy foods, rich in history, memory and tradition and cannot be replaced by a standardized diet little concerned with the construction of taste and pleasure, having a relationship with food as if it were a Medicinal product.

Keywords: food traditions, food transformations, healthy eating, medicalization of food

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Authors: Adeiza Matthew, Oluwadamilola Abubakar

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Wood energy, also known as wood fuel, is a renewable energy source that is derived from woody biomass, which is organic matter that is harvested from forests, woodlands, and other lands. Woody biomass includes trees, branches, twigs, and other woody debris that can be used as fuel. Wood energy can be classified based on its sources, such as trees outside forests, residues from wood harvesting and conversion, and energy plantations. There are several policy frameworks that support the use of wood energy, including participatory forest management and agroforestry. These policies aim to promote the sustainable use of woody biomass as a source of energy while also protecting forests and wildlife habitats. There are several options for using wood as a fuel, including central heating systems, pellet-based systems, wood chip-based systems, log boilers, fireplaces, and stoves. Each of these options has its own benefits and drawbacks, and the most appropriate option will depend on factors such as the availability of woody biomass, the heating needs of the household or facility, and the local climate. In order to use wood as a fuel, it must be harvested and stored properly. Hardwood or softwood can be used as fuel, and the heating value of firewood depends on the species of tree and the degree of moisture content. Proper harvesting and storage of wood can help to minimize environmental impacts and improve wildlife habitats. The use of wood energy has several environmental impacts, including the release of greenhouse gases during combustion and the potential for air pollution from combustion by-products. However, wood energy can also have positive environmental impacts, such as the sequestration of carbon in trees and the reduction of reliance on fossil fuels. The regulation and legislation of wood energy vary by country and region, and there is an ongoing debate about the potential use of wood energy in renewable energy technologies. Wood energy is a renewable energy source that can be used to generate electricity, heat, and transportation fuels. Woody biomass is abundant and widely available, making it a potentially significant source of energy for many countries. The use of wood energy can create local economic and employment opportunities, particularly in rural areas. Wood energy can be used to reduce reliance on fossil fuels and reduce greenhouse gas emissions. Properly managed forests can provide a sustained supply of woody biomass for energy, helping to reduce the risk of deforestation and habitat loss. Wood energy can be produced using a variety of technologies, including direct combustion, co-firing with fossil fuels, and the production of biofuels. The environmental impacts of wood energy can be minimized through the use of best practices in harvesting, transportation, and processing. Wood energy is regulated and legislated at the national and international levels, and there are various standards and certification systems in place to promote sustainable practices. Wood energy has the potential to play a significant role in the transition to a low-carbon economy and the achievement of climate change mitigation goals.

Keywords: biomass, timber, charcoal, firewood

Procedia PDF Downloads 75

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Stanislav Babaev, Vladimir Cuk, Sjef Cobben, Jan Desmet

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The procedures for performing harmonic studies for medium-voltage distribution feeders have become relatively mature topics since the early 1980s. The efforts of various electric power engineers and researchers were mainly focused on handling large harmonic non-linear loads connected scarcely at several buses of medium-voltage feeders. In order to assess the impact of these loads on the voltage quality of the distribution system, specific modeling and simulation strategies were proposed. These methodologies could deliver a reasonable estimation accuracy given the requirements of least computational efforts and reduced complexity. To uphold these requirements, certain analysis assumptions have been made, which became de facto standards for establishing guidelines for harmonic analysis. Among others, typical assumptions include balanced conditions of the study and the negligible impact of impedance frequency characteristics of various power system components. In latter, skin and proximity effects are usually omitted, and resistance and reactance values are modeled based on the theoretical equations. Further, the simplifications of the modelling routine have led to the commonly accepted practice of neglecting phase angle diversity effects. This is mainly associated with developed load models, which only in a handful of cases are representing the complete harmonic behavior of a certain device as well as accounting on the harmonic interaction between grid harmonic voltages and harmonic currents. While these modelling practices were proven to be reasonably effective for medium-voltage levels, similar approaches have been adopted for low-voltage distribution systems. Given modern conditions and massive increase in usage of residential electronic devices, recent and ongoing boom of electric vehicles, and large-scale installing of distributed solar power, the harmonics in current low-voltage grids are characterized by high degree of variability and demonstrate sufficient diversity leading to a certain level of cancellation effects. It is obvious, that new modelling algorithms overcoming previously made assumptions have to be accepted. In this work, a simulation approach aimed to deal with some of the typical assumptions is proposed. A practical low-voltage feeder is modeled in PowerFactory. In order to demonstrate the importance of diversity effect and harmonic interaction, previously developed measurement-based models of photovoltaic inverter and battery charger are used as loads. The Python-based script aiming to supply varying voltage background distortion profile and the associated current harmonic response of loads is used as the core of unbalanced simulation. Furthermore, the impact of uncertainty of feeder frequency-impedance characteristics on total harmonic distortion levels is shown along with scenarios involving linear resistive loads, which further alter the impedance of the system. The comparative analysis demonstrates sufficient differences with cases when all the assumptions are in place, and results indicate that new modelling and simulation procedures need to be adopted for low-voltage distribution systems with high penetration of non-linear loads and renewable generation.

Keywords: electric power system, harmonic distortion, power quality, public low-voltage network, harmonic modelling

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Authors: Tatiane Benvenuti, Fernando Hamerski, Alexandre Giacobbo, Andrea M. Bernardes, Marco A. S. Rodrigues

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Population growth in urban areas has caused damages to the environment, a consequence of the uncontrolled dumping of domestic and industrial wastewater. The capacity of some plants to purify domestic and agricultural wastewater has been demonstrated by several studies. Since natural wetlands have the ability to transform, retain and remove nutrients, constructed wetlands have been used for wastewater treatment. They are widely recognized as an economical, efficient and environmentally acceptable means of treating many different types of wastewater. T. domingensis Pers. species have shown a good performance and low deployment cost to extract, detoxify and sequester pollutants. Constructed Floating Wetlands (CFWs) consist of emergent vegetation established upon a buoyant structure, floating on surface waters. The upper parts of the vegetation grow and remain primarily above the water level, while the roots extend down in the water column, developing an extensive under water-level root system. Thus, the vegetation grows hydroponically, performing direct nutrient uptake from the water column. Biofilm is attached on the roots and rhizomes, and as physical and biochemical processes take place, the system functions as a natural filter. The aim of this study is to diagnose the application of macrophytes in artificial floating in the treatment of domestic sewage in south Brazil. The T. domingensis Pers. plants were placed in a flotation system (polymer structure), in full scale, in a sewage treatment plant. The sewage feed rate was 67.4 m³.d⁻¹ ± 8.0, and the hydraulic retention time was 11.5 d ± 1.3. This CFW treat the sewage generated by 600 inhabitants, which corresponds to 12% of the population served by this municipal treatment plant. During 12 months, samples were collected every two weeks, in order to evaluate parameters as chemical oxygen demand (COD), biochemical oxygen demand in 5 days (BOD5), total Kjeldahl nitrogen (TKN), total phosphorus, total solids, and metals. The average removal of organic matter was around 55% for both COD and BOD5. For nutrients, TKN was reduced in 45.9% what was similar to the total phosphorus removal, while for total solids the reduction was 33%. For metals, aluminum, copper, and cadmium, besides in low concentrations, presented the highest percentage reduction, 82.7, 74.4 and 68.8% respectively. Chromium, iron, and manganese removal achieved values around 40-55%. The use of T. domingensis Pers. in artificial floating for sewage treatment is an effective and innovative alternative in Brazilian sewage treatment systems. The evaluation of additional parameters in the treatment system may give useful information in order to improve the removal efficiency and increase the quality of the water bodies.

Keywords: constructed wetland, floating system, sewage treatment, Typha domingensis Pers.

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Authors: Leah Li Echiverri, Haoyu Shang, Yue Li

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Wenzhou-Kean University (WKU) is a Sino-US Cooperative University in China. It practices the English Immersion Program (EIP), where all the courses are taught in English. Class discussions and presentations are pervasively interwoven in designing students’ learning experiences. This WKU model has brought positive influences on students and is in some way ahead of traditional college English majors. However, literature to support the perceptions on the positive outcomes of this teaching and learning model remain scarce. The distinctive profile of Chinese-ESL students in an English Medium of Instruction (EMI) environment contributes further to the scarcity of literature compared to existing studies conducted among ESL learners in Western educational settings. Hence, the study investigated the students’ perceptions towards the English Immersion Program and determine how it influences Chinese-ESL students’ academic performance (AP). This research can provide empirical data that would be helpful to educators, teaching practitioners, university administrators, and other researchers in making informed decisions when developing curricular reforms, instructional and pedagogical methods, and university-wide support programs using this educational model. The purpose of the study was to establish the relationship between the English Immersion Program and Academic Performance among Chinese-ESL students enrolled at WKU for the academic year 2020-2021. Course length, immersion location, course type, and instructional design were the constructs of the English immersion program. English language learning, learning efficiency, and class participation were used to measure academic performance. Descriptive-correlational design was used in this cross-sectional research project. A quantitative approach for data analysis was applied to determine the relationship between the English immersion program and Chinese-ESL students’ academic performance. The research was conducted at WKU; a Chinese-American jointly established higher educational institution located in Wenzhou, Zhejiang province. Convenience, random, and snowball sampling of 283 students, a response rate of 10.5%, were applied to represent the WKU student population. The questionnaire was posted through the survey website named Wenjuanxing and shared to QQ or WeChat. Cronbach’s alpha was used to test the reliability of the research instrument. Findings revealed that when professors integrate technology (PowerPoint, videos, and audios) in teaching, students pay more attention. This contributes to the acquisition of more professional knowledge in their major courses. As to course immersion, students perceive WKU as a good place to study, providing them a high degree of confidence to talk with their professors in English. This also contributes to their English fluency and better pronunciation in their communication. In the construct of designing instruction, the use of pictures, video clips, and professors’ non-verbal communication, and demonstration of concern for students encouraged students to be more active in-class participation. Findings on course length and academic performance indicated that students’ perception regarding taking courses during fall and spring terms can moderately contribute to their academic performance. In conclusion, the findings revealed a significantly strong positive relationship between course type, immersion location, instructional design, and academic performance.

Keywords: class participation, English immersion program, English language learning, learning efficiency

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Authors: Sergio A. Bernal-Chavez, Sergio Alcalá-Alcalá, Doris A. Cerecedo-Mercado, Adriana Ganem-Rondero

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The prevalence of people with chronic wounds has increased dramatically by many factors including smoking, obesity and chronic diseases, such as diabetes, that can slow the healing process and increase the risk of becoming chronic. Because of this situation, the improvement of chronic wound treatments is a necessity, which has led to the scientific community to focus on improving the effectiveness of current therapies and the development of new treatments. The wound formation is a physiological complex process, which is characterized by an inflammatory stage with the presence of proinflammatory cells that create a proteolytic microenvironment during the healing process, which includes the degradation of important growth factors and cytokines. This decrease of growth factors and cytokines provides an interesting strategy for wound healing if they are administered externally. The use of nanometric drug delivery systems, such as polymer nanoparticles (NP), also offers an interesting alternative around dermal systems. An interesting strategy would be to propose a formulation based on a thermosensitive hydrogel loaded with polymeric nanoparticles that allows the inclusion and application of a platelet lysate (PL) on damaged skin, with the aim of promoting wound healing. In this work, NP were prepared by a double emulsion-solvent evaporation technique, using polylactic-co-glycolic acid (PLGA) as biodegradable polymer. Firstly, an aqueous solution of PL was emulsified into a PLGA organic solution, previously prepared in dichloromethane (DCM). Then, this disperse system (W/O) was poured into a polyvinyl alcohol (PVA) solution to get the double emulsion (W/O/W), finally the DCM was evaporated by magnetic stirring resulting in the NP formation containing PL. Once the NP were obtained, these systems were characterized by morphology, particle size, Z-potential, encapsulation efficiency (%EE), physical stability, infrared spectrum, calorimetric studies (DSC) and in vitro release profile. The optimized nanoparticles were included in a thermosensitive gel formulation of Pluronic® F-127. The gel was prepared by the cold method at 4 °C and 20% of polymer concentration. Viscosity, sol-gel phase transition, time of no flow solid-gel at wound temperature, changes in particle size by temperature-effect using dynamic light scattering (DLS), occlusive effect, gel degradation, infrared spectrum and micellar point by DSC were evaluated in all gel formulations. PLGA NP of 267 ± 10.5 nm and Z-potential of -29.1 ± 1 mV were obtained. TEM micrographs verified the size of NP and evidenced their spherical shape. The %EE for the system was around 99%. Thermograms and in infrared spectra mark the presence of PL in NP. The systems did not show significant changes in the parameters mentioned above, during the stability studies. Regarding the gel formulation, the transition sol-gel occurred at 28 °C with a time of no flow solid-gel of 7 min at 33°C (common wound temperature). Calorimetric, DLS and infrared studies corroborated the physical properties of a thermosensitive gel, such as the micellar point. In conclusion, the thermosensitive gel described in this work, contains therapeutic amounts of PL and fulfills the technological properties to be used in damaged skin, with potential application in wound healing and tissue regeneration.

Keywords: growth factors, polymeric nanoparticles, thermosensitive hydrogels, tissue regeneration

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Authors: Cathrine Scharning Cornwall-Nyquist, David Rafael Vaz Fernandes

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Since its origins, mankind has been dreaming of communicating directly with other animals. Past civilizations interacted on different levels with other species and recognized them in their rituals and daily activities. However, recent scientific developments have limited the ability of humans to consider deeper levels of interaction beyond observation and/or physical behavior. In recent years, animal caretakers and facilities such as sanctuaries or rescue centers have been introducing new techniques based on intuition. Most of those initiatives are related to specific cases, such as the incapacity to understand an animal’s behavior. Respected organizations also include intuitive animal communication (IAC) sessions to follow up on past interventions with their animals. Despite the lack of credibility of this discipline, some animal caring structures have opted to integrate IAC into their daily routines and approaches to animal welfare. At this stage, animal communication will be generally defined as the ability of humans to communicate with animals on an intuitive level. The trend in the field remains to be explored. The lack of theory and previous research urges the scientific community to improve the description of the phenomenon and its consequences. Considering the current scenario, qualitative approaches may become a suitable pathway to explore this topic. The purpose of this case study is to explore the beliefs behind and the consequences of an approach based on intuitive animal communication techniques for Panthera Africa (PA), an ethical sanctuary located in South Africa. Due to their personal experience, the Sanctuary’s founders have developed a philosophy based on IAC while respecting the world's highest standards for big cat welfare. Their dual approach is reflected in their rescues, daily activities, and healing animals’ trauma. The case study's main research questions will be: (i) Why do they choose to apply IAC in their work? (ii) What consequences to their activities do IAC bring? (iii) What effects do IAC techniques bring in their interactions with the outside world? Data collection will be gathered on-site via: (i) Complete participation (field notes); (ii) Semi-structured interviews (audio transcriptions); (iii) Document analysis (internal procedures and policies); (iv) Audio-visual material (communication with third parties). The main researcher shall become an active member of the Sanctuary during a 30-day period and have full access to the site. Access to documents and audio-visual materials will be granted on a request basis. Interviews are expected to be held with PA founders and staff members and with IAC practitioners related to the facility. The information gathered shall enable the researcher to provide an extended description of the phenomenon and explore its internal and external consequences for Panthera Africa.

Keywords: animal welfare, intuitive animal communication, Panthera Africa, rescue

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Authors: Palak V. Patel, Jessica Pixley, Steven R. Feldman

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Introduction: Basal cell carcinoma (BCC) is the most common skin cancer worldwide and requires substantial resources to treat. When choosing between indicated therapies, providers consider their associated adverse effects, efficacy, cosmesis, and function preservation. The patient’s tumor burden, infiltrative risk, and risk of tumor recurrence are also considered. Treatment cost is often left out of these discussions. This can lead to financial toxicity, which describes the harm and quality of life reductions inflicted by high care costs. Methods: We studied the guidelines set forth by the American Academy of Dermatology for the treatment of BCC. A PubMed literature search was conducted to identify the costs of each recommended therapy. We discuss costs alongside treatment efficacy and side-effect profile. Results: Surgical treatment for BCC can be cost-effective if the appropriate treatment is selected for the presenting tumor. Curettage and electrodesiccation can be used in low-grade, low-recurrence tumors in aesthetically unimportant areas. The benefits of cost-conscious care are not likely to be outweighed by the risks of poor cosmesis or tumor return ($471 BCC of the cheek). When tumor burden is limited, MMS offers better cure rates and lower recurrence rates than surgical excision, and with comparable costs (MMS $1263; SE $949). Surgical excision with permanent sections may be indicated when tumor burden is more extensive or if molecular testing is necessary. The utility of surgical excision with frozen sections, which costs substantially more than MMS without comparable outcomes, is less clear (SE with frozen sections $2334-$3085). Less data exists on non-surgical treatments for BCC. These techniques cost less, but recurrence-risk is high. Side-effects of nonsurgical treatment are limited to local skin reactions, and cosmesis is good. Cryotherapy, 5-FU, and MAL-PDT are all more affordable than surgery, but high recurrence rates increase risk of secondary financial and psychosocial burden (recurrence rates 21-39%; cost $100-270). Radiation therapy offers better clearance rates than other nonsurgical treatments but is associated with similar recurrence rates and a significantly larger financial burden ($2591-$3460 BCC of the cheek). Treatments for advanced or metastatic BCC are extremely costly, but few patients require their use, and the societal cost burden remains low. Vismodegib and sonidegib have good response rates but substantial side effects, and therapy should be combined with multidisciplinary care and palliative measures. Expert-review has found sonidegib to be the less expensive and more efficacious option (vismodegib $128,358; sonidegib $122,579). Platinum therapy, while not FDA-approved, is also effective but expensive (~91,435). Immunotherapy offers a new line of treatment in patients intolerant of hedgehog inhibitors ($683,061). Conclusion: Dermatologists working within resource-compressed practices and with resource-limited patients must prudently manage the healthcare dollar. Surgical therapies for BCC offer the lowest risk of recurrence at the most reasonable cost. Non-surgical therapies are more affordable, but high recurrence rates increase the risk of secondary financial and psychosocial burdens. Treatments for advanced BCC are incredibly costly, but the low incidence means the overall cost to the system is low.

Keywords: nonmelanoma skin cancer, basal cell skin cancer, squamous cell skin cancer, cost of care

Procedia PDF Downloads 101

Authors: Trevor Malambo Simbayi, Diane Hildebrandt, Tonderayi Matambo

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The rapid growth of population in recent decades has resulted in an increased need for energy to meet human activities. As energy demands increase, the need for other sources of energy other than fossil fuels, increases in turn. Furthermore, environmental concerns such as global warming due to the use of fossil fuels, depleting fossil fuel reserves and the rising cost of oil have contributed to an increased interest in renewables sources of energy. Biogas is a renewable source of energy produced through the process of anaerobic digestion (AD) and it offers a two-fold solution; it provides an environmentally friendly source of energy and its production helps to reduce the amount of organic waste taken to landfills. This research seeks to address the waste management problem caused by an aquatic weed called water hyacinth (Eichhornia crassipes) at the Hartbeespoort (Harties) Dam in the North West Province of South Africa, through biogas production of the weed. Water hyacinth is a category 1 invasive species and it is deemed to be the most problematic aquatic weed. This weed is said to double its size in the space of five days. Eutrophication in the Hartbeespoort Dam has manifested itself through the excessive algae bloom and water hyacinth infestation. A large amount of biomass from water hyacinth and algae are generated per annum from the two hundred hectare surface area of the dam exposed to the sun. This biomass creates a waste management problem. Water hyacinth when in full bloom can cover nearly half of the surface of Hartbeespoort Dam. The presence of water hyacinth in the dam has caused economic and environmental problems. Economic activities such as fishing, boating, and recreation, are hampered by the water hyacinth’s prolific growth. This research proposes the use of water hyacinth as a feedstock or substrate for biogas production in order to find an economic and environmentally friendly means of waste management for the communities living around the Hartbeespoort Dam. In order to achieve this objective, water hyacinth will be collected from the dam and it will be mechanically pretreated before anaerobic digestion. Pretreatment is required for lignocellulosic materials like water hyacinth because such materials are called recalcitrant solid materials. Cow manure will be employed as a source of microorganisms needed for biogas production to occur. Once the water hyacinth and the cow dung are mixed, they will be placed in laboratory anaerobic reactors. Biogas production will be monitored daily through the downward displacement of water. Characterization of the substrates (cow manure and water hyacinth) to determine the nitrogen, sulfur, carbon and hydrogen, total solids (TS) and volatile solids (VS). Liquid samples from the anaerobic digesters will be collected and analyzed for volatile fatty acids (VFAs) composition by means of a liquid gas chromatography machine.

Keywords: anaerobic digestion, biogas, waste management, water hyacinth

Procedia PDF Downloads 169

Authors: Roshni Raha, Karthikeyan S.

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The world's largest livestock population resides in India. Existing strategies must be modified to increase the production of livestock and their by-products in order to meet the demands of the growing human population. Even though India leads the world in both milk production and the number of cows, average production is not very healthy and productive. This may be due to the animals' poor nutrition caused by a chronic under-availability of high-quality fodder and feed. This article explores Azolla pinnata to be a promising source to produce high-quality unconventional feed and fodder for effective livestock production and good quality breeding in India. This article is an exploratory study using a literature survey and experimentation analysis. In the realm of agri-biotechnology, azolla sp gained attention for helping farmers achieve sustainability, having minimal land requirements, and serving as a feed element that doesn't compete with human food sources. It has high methionine content, which is a good source of protein. It can be easily digested as the lignin content is low. It has high antioxidants and vitamins like beta carotene, vitamin A, and vitamin B12. Using this concept, the paper aims to investigate and develop a model of using azolla plants as a novel, high-potential feed source to combat the problems of low production and poor quality of animals in India. A representative sample of animal feed is collected where azolla is added. The sample is ground into a fine powder using mortar. PITC (phenylisothiocyanate) is added to derivatize the amino acids. The sample is analyzed using HPLC (High-Performance Liquid Chromatography) to measure the amino acids and monitor the protein content of the sample feed. The amino acid measurements from HPLC are converted to milligrams per gram of protein using the method of amino acid profiling via a set of calculations. The amino acid profile data is then obtained to validate the proximate results of nutrient enhancement of the composition of azolla in the sample. Based on the proximate composition of azolla meal, the enhancement results shown were higher compared to the standard values of normal fodder supplements indicating the feed to be much richer and denser in nutrient supply. Thus azolla fed sample proved to be a promising source for animal fodder. This would in turn lead to higher production and a good breed of animals that would help to meet the economic demands of the growing Indian population. Azolla plants have no side effects and can be considered as safe and effective to be immersed in the animal feed. One area of future research could begin with the upstream scaling strategy of azolla plants in India. This could involve introducing several bioreactor types for its commercial production. Since azolla sp has been proved in this paper as a promising source for high quality animal feed and fodder, large scale production of azolla plants will help to make the process much quicker, more efficient and easily accessible. Labor expenses will also be reduced by employing bioreactors for large-scale manufacturing.

Keywords: azolla, fodder, nutrient, protein

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Authors: Rosa Palmeri, Julieta I. Monteleone, Antonio C. Barbera, Carmelo Maucieri, Aldo Todaro, Virgilio Giannone, Giovanni Spagna

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In recent years, considerable interest has been shown in the functional properties of foods, and a relevant role has been played by phenolic compounds, able to scavenge free radicals. A more sustainable agriculture has to emerge to guarantee food supply over the next years. Wheat, corn, and rice are the most common cereals cultivated, but also other cereal species, such as barley can be appreciated for their peculiarities. Barley (Hordeum vulgare L.) is a C3 winter cereal that shows high resistance at drought and salt stresses. There are growing interests in barley as ingredient for the production of functional foods due to its high content of phenolic compounds and Beta-glucans. In this respect, the possibility of separating specific functional fractions from food industry by-products looks very promising. Olive leaves represent a quantitatively significant by-product of olive grove farming, and are an interesting source of phenolic compounds. In particular, oleuropein, which provide important nutritional benefits, is the main phenolic compound in olive leaves and ranges from 17% to 23% depending upon the cultivar and growing season period. Together with oleuropein and its derivatives (e.g. dimethyloleuropein, oleuropein diglucoside), olive leaves further contain tyrosol, hydroxytyrosol, and a series of secondary metabolities structurally related to them: verbascoside, ligstroside, hydroxytyrosol glucoside, tyrosol glucoside, oleuroside, oleoside-11-methyl ester, and nuzhenide. Several flavonoids, flavonoid glycosides, and phenolic acids have also described in olive leaves. The aim of this work was the production of functional food with higher content of polyphenols and the evaluation of their shelf life. Organic durum wheat and barley grains contain higher levels of phenolic compounds were used for the production of crackers. Olive leaf extract (OLE) was obtained from cv. ‘Biancolilla’ by aqueous extraction method. Two baked goods trials were performed with both organic durum wheat and barley flours, adding olive leaf extract. Control crackers, made as comparison, were produced with the same formulation replacing OLE with water. Total phenolic compound, moisture content, activity water, and textural properties at different time of storage were determined to evaluate the shelf-life of the products. Our the preliminary results showed that the enriched crackers showed higher phenolic content and antioxidant activity than control. Alternative uses of olive leaf extracts for crackers production could represent a good candidate for the addition of functional ingredients because bakery items are daily consumed, and have long shelf-life.

Keywords: barley, functional foods, olive leaf, polyphenols, shelf life

Procedia PDF Downloads 281

Authors: Gagan Brar, Munruchi Kaur

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Macrocybe gigantea was collected from the wild, growing as pure white, fleshy, robust fruit bodies in caespitose clusters. Initially, the few ladies collecting these fruiting bodies for cooking revealed their edibility status, which was later confirmed through classical and molecular taxonomy. The culture of this potential wild edible taxa was raised with an aim of domesticating it. Various solid and liquid media were evaluated for their vegetative growth, in which Malt Extract Agar was found to be the best solid medium and Glucose Peptone medium as the best liquid medium. The effect of different temperatures as well as pH was also evaluated for the vegetative growth of M. gigantea, and it was found that it shows maximum vegetative growth at 30° and pH 5. For spawn preparation, various grains viz. Wheat grains, Jowar grains, Bajra grains and Maize grains were evaluated, and it was found that wheat grains boiled for 30 minutes gave the maximum mycelial growth. Mother spawn was thus prepared on wheat grains boiled for 30 minutes. For raising the fruiting bodies, different locally available agro-wastes were tried, and it was found that paddy straw gives the best growth. Both wilds as well as cultivated M. gigantea were compared through HPLC to evaluate the different nutritional and nutraceutical values. For the evaluation of different sugars in wild and cultivated M. gigantea, 15 sugars were taken for analysis. Among these Melezitose, Trehalose, Glucose, Xylose and Mannitol were found in the wild collection of M. gigantea; in the cultivated sample, Melezitose, Trehalose, Xylose and Dulcitol were detected. Among the 20 different amino acids, 18 amino acids were found, except Asparagine and Glutamine in both wild as well as cultivated samples. Among the 37 tested fatty acids, only 6 fatty acids, namely Palmitic acid, Stearic acid, Cis-9 Oleic acid, Linoleic acid, Gamma-Linolenic acid and Tricosanoic acid, were found in both wild and cultivated samples, although the concentration of these fatty acids was more in the cultivated sample. From the various vitamins tested, Vitamin C, D and E were present in both wild and cultivated samples. Both wild as well as cultivated samples were evaluated for the presence of phenols; for this purpose, eleven phenols were taken as standards in HPLC analysis, and it was found that Gallic acid, Resorcinol, Ferulic acid and Pyrogallol were present in the wild mushroom sample whereas in the cultivated sample Ferulic acid, Caffeic Acid, Vanillic acid and Vanillin are present. The flavonoid analysis revealed the presence of Rutin, Naringin and Quercetin in wild M. gigantea, while 5 Naringin, Catechol, Myrecetin, Gossypin and Quercetin were found in cultivated one. From the comparative chromatographic profiling of both wild as well as cultivated M. gigantea, it is concluded that no nutrient loss was found during its cultivation. An increase in percentage of secondary metabolites (i.e., phenols and flavonoids) was found in cultivated one as compared to wild M. gigantea. Thus, from future perspective cultivated species of M. gigantea can be recommended for the commercial purpose as a good food supplement.

Keywords: culture, edible, fruit bodies, wild

Procedia PDF Downloads 42

Authors: Eric Mwenda, Shadrack Ngene

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Human-wildlife conflicts (HWCs) are the main threat to conservation in Africa. This is because wildlife needs overlap with those of humans. In Kenya, about 70% of wildlife occurs outside protected areas. As a result, wildlife and human range overlap, causing HWCs. The HWCs in Kenya occur in the drylands adjacent to protected areas. The top five counties with the highest incidences of HWC are Taita Taveta, Narok, Lamu, Kajiado, and Laikipia. The common wildlife species responsible for HWCs are elephants, buffaloes, hyenas, hippos, leopards, baboons, monkeys, snakes, and crocodiles. To ensure individuals affected by the conflicts are compensated, Kenya has developed a model of HWC compensation claims data collection and payment. We collected data on HWC from all eight Kenya Wildlife Service (KWS) Conservation Areas from 2009 to 2019. Additional data was collected from stakeholders' consultative workshops held in the Conservation Areas and a literature review regarding payment of injuries and ongoing insurance schemes being practiced in areas. This was followed by the description of the claims administration process and calculation of the pricing of the compensation claims. We further developed a digital platform for data capture and processing of all reported conflict cases and payments. Our product recognized four categories of HWC (i.e., human death and injury, property damage, crop destruction, and livestock predation). Personal bodily injury and human death were provided based on the Continental Scale of Benefits. We proposed a maximum of Kenya Shillings (KES) 3,000,000 for death. Medical, pharmaceutical, and hospital expenses were capped at a maximum of KES 150,000, as well as funeral costs at KES 50,000. Pain and suffering were proposed to be paid for 12 months at the rate of KES 13,500 per month. Crop damage was to be based on farm input costs at a maximum of KES 150,000 per claim. Livestock predation leading to death was based on Tropical Livestock Unit (TLU), which is equivalent to KES 30,000, whick includes Cattle (1 TLU = KES 30,000), Camel (1.4 TLU = KES 42,000), Goat (0.15 TLU = 4,500), Sheep (0.15 TLU = 4,500), and Donkey (0.5 TLU = KES 15,000). Property destruction (buildings, outside structures and harvested crops) was capped at KES 150,000 per any one claim. We conclude that it is possible to use an administrator to collect data on HWC compensation claims and make payments using technology. The success of the new approach will depend on a piloting program. We recommended that a pilot scheme be initiated for eight months in Taita Taveta, Kajiado, Baringo, Laikipia, Narok, and Meru Counties. This will test the claims administration process as well as harmonize data collection methods. The results of this pilot will be crucial in adjusting the scheme before country-wide roll out.

Keywords: human-wildlife conflicts, compensation, human death and injury, crop destruction, predation, property destruction

Procedia PDF Downloads 30

Authors: Satam Alotibi, Muhammad J. Al-Zahrani, Fahd K. Al-Naqidan, Turki S. Hussein, Moteb Alotaibi, Mohammed Alyami, Mahdy M. Elmahdy, Abdellah Kaiba, Fatehia S. Alhakami, Talal F. Qahtan

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Water pollution is a critical global challenge that demands scalable and effective solutions for water decontamination. In this captivating research, we unveil a groundbreaking strategy for harnessing solar energy to synthesize silver (Ag) clusters on stable titanium dioxide (TiO₂) nanoparticles dispersed in water, without the need for traditional stabilization agents. These Ag-loaded TiO₂ nanoparticles exhibit exceptional photocatalytic activity, surpassing that of pristine TiO₂ nanoparticles, offering a promising solution for highly efficient water decontamination under sunlight irradiation. To the best knowledge, we have developed a unique method to stabilize TiO₂ P25 nanoparticles in water without the use of stabilization agents. This breakthrough allows us to create an ideal platform for the solar-driven synthesis of Ag clusters. Under sunlight irradiation, the stable dispersion of TiO₂ P25 nanoparticles acts as a highly efficient photocatalyst, generating electron-hole pairs. The photogenerated electrons effectively reduce silver ions derived from a silver precursor, resulting in the formation of Ag clusters. The Ag clusters loaded on TiO₂ P25 nanoparticles exhibit remarkable photocatalytic activity for water decontamination under sunlight irradiation. Acting as active sites, these Ag clusters facilitate the generation of reactive oxygen species (ROS) upon exposure to sunlight. These ROS play a pivotal role in rapidly degrading organic pollutants, enabling efficient water decontamination. To confirm the success of our approach, we characterized the synthesized Ag-loaded TiO₂ P25 nanoparticles using cutting-edge analytical techniques, such as transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and spectroscopic methods. These characterizations unequivocally confirm the successful synthesis of Ag clusters on stable TiO₂ P25 nanoparticles without traditional stabilization agents. Comparative studies were conducted to evaluate the superior photocatalytic performance of Ag-loaded TiO₂ P25 nanoparticles compared to pristine TiO₂ P25 nanoparticles. The Ag clusters loaded on TiO₂ P25 nanoparticles exhibit significantly enhanced photocatalytic activity, benefiting from the synergistic effect between the Ag clusters and TiO₂ nanoparticles, which promotes ROS generation for efficient water decontamination. Our scalable strategy for synthesizing Ag clusters on stable TiO₂ P25 nanoparticles without stabilization agents presents a game-changing solution for highly efficient water decontamination under sunlight irradiation. The use of commercially available TiO₂ P25 nanoparticles streamlines the synthesis process and enables practical scalability. The outstanding photocatalytic performance of Ag-loaded TiO₂ P25 nanoparticles opens up new avenues for their application in large-scale water treatment and remediation processes, addressing the urgent need for sustainable water decontamination solutions.

Keywords: water pollution, solar energy, silver clusters, TiO₂ nanoparticles, photocatalytic activity

Procedia PDF Downloads 43

Authors: Alper T. Celebi, Ali Beskok

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Electroosmotic (EO) slip flows in nanochannels are investigated using non-equilibrium molecular dynamics (MD) simulations, and the results are compared with analytical solution of Poisson-Boltzmann and Stokes (PB-S) equations with slip contribution. The ultimate objective of this study is to show that well-known continuum flow model can accurately predict the EO velocity profiles in nanochannels using the slip lengths and apparent viscosities obtained from force-driven flow simulations performed at various liquid-wall interaction strengths. EO flow of aqueous NaCl solution in silicon nanochannels are simulated under realistic electrochemical conditions within the validity region of Poisson-Boltzmann theory. A physical surface charge density is determined for nanochannels based on dissociations of silanol functional groups on channel surfaces at known salt concentration, temperature and local pH. First, we present results of density profiles and ion distributions by equilibrium MD simulations, ensuring that the desired thermodynamic state and ionic conditions are satisfied. Next, force-driven nanochannel flow simulations are performed to predict the apparent viscosity of ionic solution between charged surfaces and slip lengths. Parabolic velocity profiles obtained from force-driven flow simulations are fitted to a second-order polynomial equation, where viscosity and slip lengths are quantified by comparing the coefficients of the fitted equation with continuum flow model. Presence of charged surface increases the viscosity of ionic solution while the velocity-slip at wall decreases. Afterwards, EO flow simulations are carried out under uniform electric field for different liquid-wall interaction strengths. Velocity profiles present finite slips near walls, followed with a conventional viscous flow profile in the electrical double layer that reaches a bulk flow region in the center of the channel. The EO flow enhances with increased slip at the walls, which depends on wall-liquid interaction strength and the surface charge. MD velocity profiles are compared with the predictions from analytical solutions of the slip modified PB-S equation, where the slip length and apparent viscosity values are obtained from force-driven flow simulations in charged silicon nano-channels. Our MD results show good agreements with the analytical solutions at various slip conditions, verifying the validity of PB-S equation in nanochannels as small as 3.5 nm. In addition, the continuum model normalizes slip length with the Debye length instead of the channel height, which implies that enhancement in EO flows is independent of the channel height. Further MD simulations performed at different channel heights also shows that the flow enhancement due to slip is independent of the channel height. This is important because slip enhanced EO flow is observable even in micro-channels experiments by using a hydrophobic channel with large slip and high conductivity solutions with small Debye length. The present study provides an advanced understanding of EO flows in nanochannels. Correct characterization of nanoscale EO slip flow is crucial to discover the extent of well-known continuum models, which is required for various applications spanning from ion separation to drug delivery and bio-fluidic analysis.

Keywords: electroosmotic flow, molecular dynamics, slip length, velocity-slip

Procedia PDF Downloads 122

Authors: Maia Kharadze, Indira Djaparidze, Maia Vanidze, Aleko Kalandia

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Modern scientific world studies chemical components and antioxidant activity of different kinds of vines according to their breed purity and location. To our knowledge, this kind of research has not been conducted in Georgia yet. The object of our research was to study Chkhaveri vine, which is included in the oldest varieties of the Black Sea basin vine. We have studied different-altitude Chkaveri grapes, juice, and wine (half dry rose-colored produced with European technologies) and their technical markers, qualitative and quantitive composition of their biologically active compounds and their antioxidant activity. We were determining the amount of phenols using Folin-Ciocalteu reagent, Flavonoids, Catechins and Anthocyanins using Spectral method and antioxidant activity using DPPH method. Several compounds were identified using –HPLC-UV-Vis, UPLC-MS methods. Six samples of Chkhaveri species– 5, 300, 360, 380, 400, 780 meter altitudes were taken and analyzed. The sample taken from 360 m altitude is distinguished by its cluster mass (383.6 grams) and high amount of sugar (20.1%). The sample taken from the five-meter altitude is distinguished by having high acidity (0.95%). Unlike other grapes varieties, such concentration of sugar and relatively low levels of citric acid ultimately leads to Chkhaveri wine individuality. Biologically active compounds of Chkhaveri were researched in 2014, 2015, 2016. The amount of total phenols in samples of 2016 fruit varies from 976.7 to 1767.0 mg/kg. Amount of Anthocians is 721.2-1630.2 mg/kg, and the amount of Flavanoids varies from 300.6 to 825.5 mg/kg. Relatively high amount of anthocyanins was found in the Chkhaveri at 780-meter altitude - 1630.2 mg/kg. Accordingly, the amount of Phenols and Flavanoids is high- 1767.9 mg/kg and 825.5 mg/kg. These characteristics are low in samples gathered from 5 meters above sea level, Anthocyanins-721.2 mg/ kg, total Phenols-976.7 mg/ kg, and Flavanoids-300.6 mg/kg. The highest amount of bioactive compounds can be found in the Chkhaveri samples of high altitudes because with rising height environment becomes harsh, the plant has to develop a better immune system using Phenolic compounds. The technology that is used for the production of wine also plays a huge role in the composition of the final product. Optimal techniques of maceration and ageing were worked out. While squeezing Chkhaveri, there are no anthocyanins in the juice. However, the amount of Anthocyanins rises during maceration. After the fermentation of dregs, the amount of anthocyanins is 55%, 521.3 gm/l, total Phenols 80% 1057.7 mg/l and Flavanoids 23.5 mg/l. Antioxidant activity of samples was also determined using the effect of 50% inhibition of the samples. All samples have high antioxidant activity. For instance, in samples at 780 meters above the sea-level antioxidant activity was 53.5%. It is relatively high compared to the sample at 5 m above sea-level with the antioxidant activity of 30.5%. Thus, there is a correlation between the amount Anthocyanins and antioxidant activity. The designated project has been fulfilled by financial support of the Georgia National Science Foundation (Grant AP/96/13, Grant 216816), Any idea in this publication is possessed by the author and may not represent the opinion of the Georgia National Science Foundation.

Keywords: antioxidants, bioactive content, wine, chkhaveri

Procedia PDF Downloads 204

Authors: Dimitra Das, Anuradha Mitra, Kalyan Kumar Chattopadhyay

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Organic polymers, constructed from light elements like carbon, hydrogen, nitrogen, oxygen, sulphur, and boron atoms, are the emergent class of non-toxic, metal-free, environmental benign advanced materials. Covalent triazine-based polymers with a functional triazine group are significant class of organic materials due to their remarkable stability arising out of strong covalent bonds. They can conventionally form hydrogen bonds, favour π–π contacts, and they were recently revealed to be involved in interesting anion–π interactions. The present work mainly focuses upon the development of a single-crystalline, highly cross-linked triazine-based nitrogen-rich organic polymer with nanodendritic morphology and significant thermal stability. The polymer has been synthesized through hydrothermal treatment of melamine and ethylene glycol resulting in cross-polymerization via condensation-polymerization reaction. The crystal structure of the polymer has been evaluated by employing Rietveld whole profile fitting method. The polymer has been found to be composed of monoclinic melamine having space group P21/a. A detailed insight into the chemical structure of the as synthesized polymer has been elucidated by Fourier Transform Infrared Spectroscopy (FTIR) and Raman spectroscopic analysis. X-Ray Photoelectron Spectroscopic (XPS) analysis has also been carried out for further understanding of the different types of linkages required to create the backbone of the polymer. The unique rod-like morphology of the triazine based polymer has been revealed from the images obtained from Field Emission Scanning Electron Microscopy (FESEM) and Transmission Electron Microscopy (TEM). Interestingly, this polymer has been found to selectively detect mercury (Hg²⁺) ions at an extremely low concentration through fluorescent quenching with detection limit as low as 0.03 ppb. The high toxicity of mercury ions (Hg²⁺) arise from its strong affinity towards the sulphur atoms of biological building blocks. Even a trace quantity of this metal is dangerous for human health. Furthermore, owing to its small ionic radius and high solvation energy, Hg²⁺ ions remain encapsulated by water molecules making its detection a challenging task. There are some existing reports on fluorescent-based heavy metal ion sensors using covalent organic frameworks (COFs) but reports on mercury sensing using triazine based polymers are rather undeveloped. Thus, the importance of ultra-trace detection of Hg²⁺ ions with high level of selectivity and sensitivity has contemporary significance. A plausible sensing phenomenon by the polymer has been proposed to understand the applicability of the material as a potential sensor. The impressive sensitivity of the polymer sample towards Hg²⁺ is the very first report in the field of highly crystalline triazine based polymers (without the introduction of any sulphur groups or functionalization) towards mercury ion detection through photoluminescence quenching technique. This crystalline metal-free organic polymer being cheap, non-toxic and scalable has current relevance and could be a promising candidate for Hg²⁺ ion sensing at commercial level.

Keywords: fluorescence quenching , mercury ion sensing, single-crystalline, triazine-based polymer

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Authors: Pakize Ozlem Kurt Polat

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GMO (genetically modified organism) is the result of a laboratory process where genes from the DNA of one species are extracted and artificially forced into the genes of an unrelated plant or animal. This technology includes; nucleic acid hybridization, recombinant DNA, RNA, PCR, cell culture and gene cloning techniques. The studies are divided into three groups of properties transferred to the transgenic plant. Up to 59% herbicide resistance characteristic of the transfer, 28% resistance to insects and the virus seems to be related to quality characteristics of 13%. Transgenic crops are not included in the commercial production of each product; mostly commercial plant is soybean, maize, canola, and cotton. Day by day increasing GMO interest can be listed as follows; Use in the health area (Organ transplantation, gene therapy, vaccines and drug), Use in the industrial area (vitamins, monoclonal antibodies, vaccines, anti-cancer compounds, anti -oxidants, plastics, fibers, polyethers, human blood proteins, and are used to produce carotenoids, emulsifiers, sweeteners, enzymes , food preservatives structure is used as a flavor enhancer or color changer),Use in agriculture (Herbicide resistance, Resistance to insects, Viruses, bacteria, fungi resistance to disease, Extend shelf life, Improving quality, Drought , salinity, resistance to extreme conditions such as frost, Improve the nutritional value and quality), we explain all this methods step by step in this research. GMO has advantages and disadvantages, which we explain all of them clearly in full text, because of this topic, worldwide researchers have divided into two. Some researchers thought that the GMO has lots of disadvantages and not to be in use, some of the researchers has opposite thought. If we look the countries law about GMO, we should know Biosafety law for each country and union. For this Biosecurity reasons, the problems caused by the transgenic plants, including Turkey, to minimize 130 countries on 24 May 2000, ‘the United Nations Biosafety Protocol’ signed nudes. This protocol has been prepared in addition to Cartagena Biosafety Protocol entered into force on September 11, 2003. This protocol GMOs in general use by addressing the risks to human health, biodiversity and sustainable transboundary movement of all GMOs that may affect the prevention, transit covers were dealt and used. Under this protocol we have to know the, ‘US Regulations GMO’, ‘European Union Regulations GMO’, ‘Turkey Regulations GMO’. These three different protocols have different applications and rules. World population increasing day by day and agricultural fields getting smaller for this reason feeding human and animal we should improve agricultural product yield and quality. Scientists trying to solve this problem and one solution way is molecular biotechnology which is including the methods of GMO too. Before decide to support or against the GMO, should know the GMO protocols and it effects.

Keywords: biotechnology, GMO (genetically modified organism), molecular marker

Procedia PDF Downloads 218

Authors: Mukul Sharma, Madhusmita Das, Sundeep Chaitanya Vedithi

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Leprosy is a chronic human disease caused by Mycobacterium leprae, that cannot be cultured in vitro. Though treatable with multidrug therapy (MDT), recently, bacteria reported resistance to multiple antibiotics. Targeting DNA replication and repair pathways can serve as the foundation of developing new anti-leprosy drugs. Due to the absence of an axenic culture medium for the propagation of M. leprae, studying cellular processes, especially those belonging to DNA repair pathways, is challenging. Genomic understanding of M. Leprae harbors several protein-coding genes with no previously assigned function known as 'hypothetical proteins'. Here, we report identification and expression of known and hypothetical DNA repair genes from a human skin biopsy and mouse footpads that are involved in base excision repair, direct reversal repair, and SOS response. Initially, a bioinformatics approach was employed based on sequence similarity, identification of known protein domains to screen the hypothetical proteins in the genome of M. leprae, that are potentially related to DNA repair mechanisms. Before testing on clinical samples, pure stocks of bacterial reference DNA of M. leprae (NHDP63 strain) was used to construct standard graphs to validate and identify lower detection limit in the qPCR experiments. Primers were designed to amplify the respective transcripts, and PCR products of the predicted size were obtained. Later, excisional skin biopsies of newly diagnosed untreated, treated, and drug resistance leprosy cases from SIHR & LC hospital, Vellore, India were taken for the extraction of RNA. To determine the presence of the predicted transcripts, cDNA was generated from M. leprae mRNA isolated from clinically confirmed leprosy skin biopsy specimen across all the study groups. Melting curve analysis was performed to determine the integrity of the amplification and to rule out primer‑dimer formation. The Ct values obtained from qPCR were fitted to standard curve to determine transcript copy number. Same procedure was applied for M. leprae extracted after processing a footpad of nude mice of drug sensitive and drug resistant strains. 16S rRNA was used as positive control. Of all the 16 genes involved in BER, DR, and SOS, differential expression pattern of the genes was observed in terms of Ct values when compared to human samples; this was because of the different host and its immune response. However, no drastic variation in gene expression levels was observed in human samples except the nth gene. The higher expression of nth gene could be because of the mutations that may be associated with sequence diversity and drug resistance which suggests an important role in the repair mechanism and remains to be explored. In both human and mouse samples, SOS system – lexA and RecA, and BER genes AlkB and Ogt were expressing efficiently to deal with possible DNA damage. Together, the results of the present study suggest that DNA repair genes are constitutively expressed and may provide a reference for molecular diagnosis, therapeutic target selection, determination of treatment and prognostic judgment in M. leprae pathogenesis.

Keywords: DNA repair, human biopsy, hypothetical proteins, mouse footpads, Mycobacterium leprae, qPCR

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Authors: Denise De Fátima Fernandes Barbosa, Tyane Mayara Ferreira Oliveira, Diego Jorge Maia Lima, Paula Renata Amorim Lessa, Ana Karina Bezerra Pinheiro, Cintia Gondim Pereira Calou, Glauberto Da Silva Quirino, Hellen Lívia Oliveira Catunda, Tatiana Gomes Guedes, Nicolau Da Costa

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The aim of this study is to evaluate the use of Digital Cervicography (DC) in the diagnosis of precancerous lesions related to Human Papillomavirus (HPV). Cross-sectional study with a quantitative approach, of evaluative type, held in a health unit linked to the Pro Dean of Extension of the Federal University of Ceará, in the period of July to August 2015 with a sample of 33 women. Data collecting was conducted through interviews with enforcement tool. Franco (2005) standardized the technique used for DC. Polymerase Chain Reaction (PCR) was performed to identify high-risk HPV genotypes. DC were evaluated and classified by 3 judges. The results of DC and PCR were classified as positive, negative or inconclusive. The data of the collecting instruments were compiled and analyzed by the software Statistical Package for Social Sciences (SPSS) with descriptive statistics and cross-references. Sociodemographic, sexual and reproductive variables were analyzed through absolute frequencies (N) and their respective percentage (%). Kappa coefficient (κ) was applied to determine the existence of agreement between the DC of reports among evaluators with PCR and also among the judges about the DC results. The Pearson's chi-square test was used for analysis of sociodemographic, sexual and reproductive variables with the PCR reports. It was considered statistically significant (p<0.05). Ethical aspects of research involving human beings were respected, according to 466/2012 Resolution. Regarding the socio-demographic profile, the most prevalent ages and equally were those belonging to the groups 21-30 and 41-50 years old (24.2%). The brown color was reported in excess (84.8%) and 96.9% out of them had completed primary and secondary school or studying. 51.5% were married, 72.7% Catholic, 54.5% employed and 48.5% with income between one and two minimum wages. As for the sexual and reproductive characteristics, prevailed heterosexual (93.9%) who did not use condoms during sexual intercourse (72.7%). 51.5% had a previous history of Sexually Transmitted Infection (STI), and HPV the most prevalent STI (76.5%). 57.6% did not use contraception, 78.8% underwent examination Cancer Prevention Uterus (PCCU) with shorter time interval or equal to one year, 72.7% had no cases of Cervical Cancer in the family, 63.6% were multiparous and 97% were not vaccinated against HPV. DC identified good level of agreement between raters (κ=0.542), had a specificity of 77.8% and sensitivity of 25% when compared their results with PCR. Only the variable race showed a statistically significant association with CRP (p=0.042). DC had 100% acceptance amongst women in the sample, revealing the possibility of other experiments in using this method so that it proves as a viable technique. The DC positivity criteria were developed by nurses and these professionals also perform PCCU in Brazil, which means that DC can be an important complementary diagnostic method for the appreciation of these professional’s quality of examinations.

Keywords: gynecological examination, human papillomavirus, nursing, papillomavirus infections, uterine lasmsneop

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Authors: Sundru Manjulata Devi, Prakash M. Halami

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The immemorial use of fermented foods from vegetables, dairy and other biological sources are of great demand in India because of their health benefits. However, the diversity of Lactobacillus plantarum group (LPG) of vegetable origin has not been revealed yet, particularly with reference to their probiotic functionalities. In the present study, the different species of probiotic Lactobacillus plantarum group (LPG) i.e., L. plantarum subsp. plantarum MTCC 5422 (from fermented cereals), L. plantarum subsp. argentoratensis FG16 (from fermented bamboo shoot) and L. paraplantarum MTCC 9483 (from fermented gundruk) (as characterized by multiplex recA PCR assay) were considered to investigate their relative expression of MUB domains of mub gene (mucin binding protein) by Real time PCR. Initially, the allelic variation in the mub gene was assessed and found to encode three different variants (Type I, II and III). All the three types had 8, 9 and 10 MUB domains respectively (as analysed by Pfam database) and were found to be responsible for adhesion of bacteria to the host intestinal epithelial cells. These domains either get inserted or deleted during speciation or evolutionary events and lead to divergence. The reverse transcriptase qPCR analysis with mubLPF1+R1 primer pair supported variation in amplicon sizes with 300, 500 and 700 bp among different LPG strains. The relative expression of these MUB domains significantly unregulated in the presence of 1% mucin in overnight grown cultures. Simultaneously, the mub gene expressed efficiently by 7 fold in the culture L. paraplantarum MTCC 9483 with 10 MUB domains. An increase in the expression levels for L. plantarum subsp. plantarum MTCC 5422 and L. plantarum subsp. argentoratensis FG16 (MCC 2974) with 9 and 8 repetitive domains was around 4 and 2 fold, respectively. The detection and expression of an integrase (int) gene in the upstream region of mub gene reveals the excision and integration of these repetitive domains. Concurrently, an in vitro adhesion assay to mucin and exclusion of pathogens (such as Listeria monocytogenes and Micrococcus leuteus) was investigated and observed that the L. paraplantarum MTCC 9483 with more adhesion domains has more ability to adhere to mucin and inhibited the growth of pathogens. The production and expression of plantaricin-like bacteriocin (plnNC8 type) in MTCC 9483 suggests the pathogen inhibition. Hence, the expression of MUB domains can act as potential biomarkers in the screening of a novel probiotic LPG strain with adherence property. The present study provides a platform for an easy, rapid, less time consuming, low-cost methodology for the detection of potential probiotic bacteria. It was known that the traditional practices followed in the preparation of fermented bamboo shoots/gundruk/cereals of Indian foods contain different kinds of neutraceuticals for functional food and novel compounds with health promoting factors. In future, a detailed study of these food products can add more nutritive value, consumption and suitable for commercialization.

Keywords: adhesion gene, fermented foods, MUB domains, probiotics

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Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have played an indispensable role in controlling illness and death associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global healthcare problem. Many antibiotics had lost their effectiveness since the beginning of the antibiotic era because many bacteria have adapted defenses against these antibiotics. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing require the isolation of the pathogen from a clinical specimen by culturing on the appropriate media (this culturing stage lasts 24 h-first culturing). Then, chosen colonies are grown on media containing antibiotic(s), using micro-diffusion discs (second culturing time is also 24 h) in order to determine its bacterial susceptibility. Other methods, genotyping methods, E-test and automated methods were also developed for testing antimicrobial susceptibility. Most of these methods are expensive and time-consuming. Fourier transform infrared (FTIR) microscopy is rapid, safe, effective and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria; nonetheless, its true potential in routine clinical diagnosis has not yet been established. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The UTI E.coli bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 700 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 90% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E.coli, FTIR, multivariate analysis, susceptibility, UTI

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Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

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Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

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Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson

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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.

Keywords: cell migration, hMSCs, SiHA, transwell migration system

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