World Academy of Science, Engineering and Technology

Authors: D. Bhowmik, Y. Tian, Q. Yin, F. Zhu

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Cyclic GMP-AMP synthase (cGAS), recognises cytoplasmic double-stranded DNA (dsDNA), indicative of bacterial and viral infections, as well as the leakage of self DNA by cellular dysfunction and stresses, to elicit the host's immune responses. Viruses also have developed numerous strategies to antagonize the cGAS-STING pathway. Kaposi's sarcoma-associated herpesvirus (KSHV) is a human DNA tumor virus that is the causative agent of Kaposi’s sarcoma and several other malignancies. To persist in the host, consequently causing diseases, KSHV must overcome the host innate immune responses, including the cGAS-STING DNA sensing pathway. We already found that ORF52 or KicGAS (KSHV inhibitor of cGAS), an abundant and basic gamma herpesvirus-conserved tegument protein, directly inhibits cGAS enzymatic activity. To better understand the mechanism, we have performed the biochemical and structural characterization of full-length KicGAS and various mutants in regarding binding to DNA. We observed that KicGAS is capable of self-association and identified the critical residues involved in the oligomerization process. We also characterized the DNA-binding of KicGAS and found that KicGAS cooperatively oligomerizes along the length of the double stranded DNA, the highly conserved basic residues at the c-terminal disordered region are crucial for DNA recognition. Deficiency in oligomerization also affects DNA binding. Thus DNA binding by KicGAS sequesters DNA and prevents it from being detected by cGAS, consequently inhibiting cGAS activation. KicGAS homologues also inhibit cGAS efficiently, suggesting inhibition of cGAS is evolutionarily conserved mechanism among gamma herpesvirus. These results highlight the important viral strategy to evade this innate immune sensor.

Keywords: Kaposi's sarcoma-associated herpesvirus, KSHV, cGAS, DNA binding, inhibition

Procedia PDF Downloads 107

Authors: Jianfang Wang, Xianzhe Chen, Zhuoliang Liu, Cheng-An Tao, Yujiao Li

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Organophosphorus (OPs) pesticide used as insecticides are widely used in agricultural pest control, household and storage deworming. The detection of pesticides needs more simple and efficient methods. One of the best ways is to make electrochemical biosensors. In this paper, an electrochemical enzyme biosensor based on acetylcholine esterase (AChE) was constructed, and its sensing properties and sensing mechanisms were studied. Reduced graphene oxide-polydopamine complexes (RGO-PDA), gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were prepared firstly and composited with AChE and chitosan (CS), then fixed on the glassy carbon electrode (GCE) surface to construct the biosensor GCE/RGO-PDA-AuNPs-AgNPs-AChE-CS by one-pot method. The results show that graphene oxide (GO) can be reduced by dopamine (DA) and dispersed well in RGO-PDA complexes. And the composites have a synergistic catalysis effect and can improve the surface resistance of GCE. The biosensor selectively can detect acetylcholine (ACh) and OPs pesticide with good linear range and high sensitivity. The performance of the biosensor is affected by the ratio and adding ways of AChE and the adding of AuNPs and AChE. And the biosensor can achieve a detection limit of 2.4 ng/L for methyl parathion and a wide linear detection range of 0.02 ng/L ~ 80 ng/L, and has excellent stability, good anti-interference ability, and excellent preservation performance, indicating that the sensor has practical value.

Keywords: acetylcholine esterase, electrochemical biosensor, nanoparticles, organophosphates, reduced graphene oxide

Procedia PDF Downloads 84

Authors: M. Z. Mahmood, S. Ismail

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A reusable immobilized unilayer thin coating of adsorbent material bentonite and photocatalyst (TiO₂) was fabricated on the glass beaker to remove aqueous methylene blue solution. The dye removal efficiency of photocatalyst was much lower with pure titanium dioxide. In the preliminary experiments, different compositions of TiO₂ – bentonite were tested on unilayer and bilayer system, and it was observed that 0.50:0.50 ratios are best for maximum photocatalytic degradation of methylene blue in aqueous medium when applied on unilayer coating system.

Keywords: adsorption, photocatalyst, bentonite, TiO₂

Procedia PDF Downloads 76

Authors: Ayesha Imtiaz, Darlina Md. Naim

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Family Nemipteridae is among the most abundantly distributed family in Malaysian fish markets due to its high contribution to landing sites of Malaysia. Using an advanced molecular approach that used two mitochondrial (Cytochrome oxidase c I and Cytochrome oxidase b) and one nuclear gene (Recombination activating gene, RAGI) to expose cryptic diversity and phylogenetic relationships among commercially important species of family Nemipteridae. Our research covered all genera (including 31 species out total 45 species) of family Nemipteridae, distributed in Malaysia. We also found certain type of geographical barriers in the South China sea that reduces dispersal and stops a few species to intermix. Northside of the South China Sea (near Vietnam) does not allow genetic diversity to mix with the Southern side of the South China sea (Sarawak) and reduces dispersal. Straits of Malacca reduce the intermixing genetic diversity of South China Sea and the Indian Ocean.

Keywords: Nemipteridae, RAG I, south east Asia, Malaysia

Procedia PDF Downloads 119

Authors: Othman Elmahdy Othman

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The objectives of this study were to identify polymorphisms in the STAT5A using Restriction Fragment Length Polymorphism and DGAT1 using Single-Strand Conformation Polymorphism genes among three Egyptian goat breeds (Barki, Zaraibi, and Damascus) as well as investigate the effect of their genotypes on milk composition traits of Zaraibi goats. One hundred and fifty blood samples were collected for DNA extraction, 60 from Zaraibi, 40 from Damascus and 50 from Barki breeds. Fat, protein and lactose percentages were determined in Zaraibi goat milk using an automatic milk analyzer. Two genotypes, CC and CT (for STAT5A) and C-C- and C-C+ (for DGAT1), were identified in the three Egyptian goat breeds with different frequencies. The associations between these genotypes and milk fat, protein and lactose were determined in Zaraibi breed. The results showed that the STAT5A genotypes had significant effects on milk yield, protein, fat and lactose with the superiority of CT genotype over CC. Regarding DGAT1 polymorphism, the result showed the only association between it with milk fat where the animals with C-C+ genotype had greater milk fat than animals possess C-C- genotype. The association of combined genotypes with milk trait declared that the does with heterozygous genotypes for both genes are preferred than does with homozygous genotypes where the animals with CTC-C+ have more milk yield, fat and protein than those with CCC-C- genotype. In conclusion, the result showed that C/T and C-/C+ SNPs of STAT5A and DGAT1 genes respectively may be useful markers for assisted selection programs to improve goat milk composition

Keywords: DGAT1, genetic polymorphism, milk trait, STAT5A

Procedia PDF Downloads 121

Authors: Syeda Fahria Hoque Mimmi

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Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.

Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk

Procedia PDF Downloads 141

Authors: Ahmed Raza Khan

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Local-strain engineering is an exciting approach to tune the optoelectronic properties of materials and enhance the performance of devices. Two dimensional (2D) materials such as 2D transition metal dichalcogenides (TMDCs) are particularly well-suited for this purpose because they have high flexibility and can withstand high deformations before rupture. Wrinkles on thick TMDC layers have been reported to show the interesting photoluminescence enhancement due to bandgap modulation and funneling effect. However, the wrinkles in ultrathin TMDCs have not been investigated, because the wrinkles can easily fall down to form folds in these ultrathin layers of TMDCs. Here, we have achieved both wrinkle and fold nano-structures simultaneously on 1-3L WS₂ using a new fabrication technique. The comparable layer dependent reduction in surface potential is observed for both folded layers and corresponding perfect pack layers due to the dominant interlayer screening effect. The strains produced from the wrinkle nanostructures considerably vary semi conductive junction properties. Thermo-ionic modelling suggests that the strained (1.6%) wrinkles can lower the Schottky barrier height (SBH) by 20%. The photo-generated carriers would further significantly lower the SBH. These results present an important advance towards controlling the optoelectronic properties of atomically thin WS₂ using strain engineering, with important implications for practical device applications.

Keywords: strain engineering, folding, WS₂, Kelvin probe force microscopy, KPFM, surface potential, photo current, layer dependence

Procedia PDF Downloads 85

Authors: Cathal Merz, Gareth O’Donnell

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Coil reinforced thin-walled (CRTW) tubes are used in medicine to treat problems affecting blood vessels within the body through minimally invasive procedures. The CRTW tube considered in this research makes up part of such a device and is inserted into the patient via their femoral or brachial arteries and manually navigated to the site in need of treatment. This procedure replaces the requirement to perform open surgery but is limited by reduction of blood vessel lumen diameter and increase in tortuosity of blood vessels deep in the brain. In order to maximize the capability of these procedures, CRTW tube devices are being manufactured with decreasing wall thicknesses in order to deliver treatment deeper into the body and to allow passage of other devices through its inner diameter. This introduces significant stresses to the device materials which have resulted in an observed increase in the breaking of the proximal segment of the device into two separate pieces after it has failed by buckling. As there is currently no international standard for measuring the mechanical properties of these CRTW tube devices, it is difficult to accurately analyze this problem. The aim of the current work is to address this discrepancy in the biomedical device industry by developing a measurement system that can be used to quantify the effect of process and design changes on CRTW tube performance, aiding in the development of better performing, next generation devices. Using materials testing frames, micro-computed tomography (micro-CT) imaging, experiment planning, analysis of variance (ANOVA), T-tests and regression analysis, test methods have been developed for assessing the impact of process and design changes on the device. The major findings of this study have been an insight into the suitability of buckle and three-point bend tests for the measurement of the effect of varying processing factors on the device’s performance, and guidelines for interpreting the output data from the test methods. The findings of this study are of significant interest with respect to verifying and validating key process and design changes associated with the device structure and material condition. Test method integrity evaluation is explored throughout.

Keywords: neurovascular catheter, coil reinforced tube, buckling, three-point bend, tensile

Procedia PDF Downloads 90

Authors: Shymaa S. Zaher, Hesham M. Abd El-Fatah, Dina M. Ali

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Burullus Lagoon is the second largest lake, along the Mediterranean seashore. It exposed to over nutrient enrichment from fish farming and agricultural drainage wastes. This study assesses the present status phytoplankton response to different flow events, including domestic, agricultural, industrial, and fish farms discharge in the three main sectors of Burullus Lagoon, to focus on the influence of environmental variables on phytoplankton species composition inhabiting the Lagoon. Twelve sites representing the eastern, central, and western basin were selected during winter and summer 2018. Among the most abundant group, Chlorophyceae came in the first rank by 37.9% of the total phytoplankton densities, Bacillariophyceae (29.31%), Cyanophyceae (20.7%), Euglenophyceae (8.63%) and Dinophyceae (3.4%). Cyclotella menenghiana was the most abundant diatoms, while Scenedesmus quadricauda, S. acuminatus, and S. bijuga were highly recorded nearby the drains (in the middle sector). Phytoplankton in Burullus Lagoon attained the lowest values during the winter season and the highest ones during the summer season. The total count of phytoplankton in the middle and western basin of the lake was higher than that of the eastern part. Excessive use of chemical fertilizers, pesticides, and washing out of nutrients loaded to the drainage water, leading to a significant pronounced decrease in community composition and standing crop of phytoplankton in Burullus Lake from year to year, hold the danger of shifting the lagoon ecosystem.

Keywords: Burullus Lagoon, environmental variables, phytoplankton, water pollution

Procedia PDF Downloads 85

Authors: M. Jerold, V. Sivasubramanian, A. George, B.S. Ashik, S. S. Kumar

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Bioethanol is a sustainable biofuel that can be used alternative to fossil fuels. Today, third generation (3G) biofuel is gaining more attention than first and second-generation biofuel. The more lignin content in the lignocellulosic biomass is the major drawback of second generation biofuels. Algae are the renewable feedstock used in the third generation biofuel production. Algae contain a large number of carbohydrates, therefore it can be used for the fermentation by hydrolysis process. There are two groups of Algae, such as micro and macroalgae. In the present investigation, Macroalgae was chosen as raw material for the production of bioethanol. Two marine algae viz. Ulva Lactuca and Sargassum swartzii were used for the experimental studies. The algal biomass was characterized using various analytical techniques like Elemental Analysis, Scanning Electron Microscopy Analysis and Fourier Transform Infrared Spectroscopy to understand the physio-Chemical characteristics. The batch experiment was done to study the hydrolysis and operation parameters such as pH, agitation, fermentation time, inoculum size. The saccharification was done with acid and alkali treatment. The experimental results showed that NaOH treatment was shown to enhance the bioethanol. From the hydrolysis study, it was found that 0.5 M Alkali treatment would serve as optimum concentration for the saccharification of polysaccharide sugar to monomeric sugar. The maximum yield of bioethanol was attained at a fermentation time of 9 days. The inoculum volume of 1mL was found to be lowest for the ethanol fermentation. The agitation studies show that the fermentation was higher during the process. The percentage yield of bioethanol was found to be 22.752% and 14.23 %. The elemental analysis showed that S. swartzii contains a higher carbon source. The results confirmed hydrolysis was not completed to recover the sugar from biomass. The specific gravity of ethanol was found to 0.8047 and 0.808 for Ulva Lactuca and Sargassum swartzii, respectively. The purity of bioethanol also studied and found to be 92.55 %. Therefore, marine algae can be used as a most promising renewable feedstock for the production of bioethanol.

Keywords: algae, biomass, bioethaol, biofuel, pretreatment

Procedia PDF Downloads 131

Authors: Franciele G. Esteves, Jose R. A. dos Santos-Pinto, Caroline L. de Souza, Mario S. Palma

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Orb-weaving spiders use a very strong, stickiness, and elastic web to catch the prey. These web properties would be enough for the entrapment of prey; however, these spiders may be hiding venomous secrets on the web, which are being revealed now. Here we provide strong proteome, peptidome, and transcriptomic evidence for the presence of toxic components on the web silk from Nephila clavipes. Our scientific outcomes revealed, both in the web silk and in the silk-producing glands, a wide diversity of toxins/neurotoxins, defensins, and proteolytic enzymes. These toxins/neurotoxins are similar to toxins isolated from animal venoms, such as Sphigomyelinase D, Latrotoxins, Zodatoxins, Ctenitoxin Pn and Pk, Agatoxins and Theraphotoxin. Moreover, the insect-toxicity results with the web silk crude extract demonstrated that these toxic components can be lethal and/or cause paralytic effects to the prey. Therefore, through OMICs approaches, the results presented until now may contribute to a better understanding of the chemical and ecological interaction of these compounds in insect-prey capture by spider web N. clavipes, demonstrating that the web is not only a simple mechanical tool but has a chemical-active involvement in prey capture. Moreover, the results can also contribute to future studies of possible development of a selective insecticide or even in possible pharmacological applications.

Keywords: web silk toxins, silk-produncing glands, de novo transcriptome assembly, LCMS-based proteomics

Procedia PDF Downloads 104

Authors: Ghaneshree Moonsamy, Nodumo N. Zulu, Rajesh Lalloo, Suren Singh, Santosh O. Ramchuran

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The abalone industry of South Africa is under severe pressure due to illegal harvesting and poaching of this seafood delicacy. These abalones are harvested excessively; as a result, these animals do not have a chance to replace themselves in their habitats, ensuing in a drastic decrease in natural stocks of abalone. Abalone has an extremely slow growth rate and takes approximately four years to reach a size that is market acceptable; therefore, it was imperative to investigate methods to boost the overall growth rate and immunity of the animal. The University of Cape Town (UCT) began to research, which resulted in the isolation of two microorganisms, a yeast isolate Debaryomyces hansenii and a bacterial isolate Vibrio midae, from the gut of the abalone and characterised them for their probiotic abilities. This work resulted in an internationally competitive concept technology that was patented. The next stage of research was to develop a suitable bioprocess to enable commercial production. Numerous steps were taken to develop an efficient production process for V. midae, one of the isolates found by UCT. The initial stages of research involved the development of a stable and robust inoculum and the optimization of physiological growth parameters such as temperature and pH. A range of temperature and pH conditions were evaluated, and data obtained revealed an optimum growth temperature of 30ᵒC and a pH of 6.5. Once these critical growth parameters were established further media optimization studies were performed. Corn steep liquor (CSL) and high test molasses (HTM) were selected as suitable alternatives to more expensive, conventionally used growth medium additives. The optimization of CSL (6.4 g.l⁻¹) and HTM (24 g.l⁻¹) concentrations in the growth medium resulted in a 180% increase in cell concentration, a 5716-fold increase in cell productivity and a 97.2% decrease in the material cost of production in comparison to conventional growth conditions and parameters used at the onset of the study. In addition, a stable market-ready liquid probiotic product, encompassing the viable but not culturable (VBNC) state of Vibrio midae cells, was developed during the downstream processing aspect of the study. The demonstration of this technology at a full manufacturing scale has further enhanced the attractiveness and commercial feasibility of this production process.

Keywords: probiotics, abalone aquaculture, bioprocess technology, manufacturing scale technology development

Procedia PDF Downloads 126

Authors: Fatima Mokeddem

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The phonocardiogram signal (PCG) is a physiological signal that reflects heart mechanical activity, is a promising tool for curious researchers in this field because it is full of indications and useful information for medical diagnosis. PCG segmentation is a basic step to benefit from this signal. Therefore, this paper presents an algorithm that serves the separation of heart sounds and heart murmurs in case they exist in order to use them in several applications and heart sounds analysis. The separation process presents here is founded on three essential steps filtering, envelope detection, and heart sounds segmentation. The algorithm separates the PCG signal into S1 and S2 and extract cardiac murmurs.

Keywords: phonocardiogram signal, filtering, Envelope, Detection, murmurs, heart sounds

Procedia PDF Downloads 112

Authors: M. Jerold, V. Sivasubramanian

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In this study, nano zero valent iron Sargassum swartzii (nZVI-SS) biocomposite a marine algal based biosorbent was used for the removal of simulated crystal violet (CV) in batch and continuous fixed bed operation. The Box-Behnen design (BBD) experimental results revealed the biosoprtion was maximum at pH 7.5, biosorbent dosage 0.1 g/L and initial CV concentration of 100 mg/L. The effect of various column parameters like bed depth (3, 6 and 9 cm), flow rate (5, 10 and 15 mL/min) and influent CV concentration (5, 10 and 15 mg/L) were investigated. The exhaustion time increased with increase of bed depth, influent CV concentration and decrease of flow rate. Adam-Bohart, Thomas and Yoon-Nelson models were used to predict the breakthrough curve and to evaluate the model parameters. Out of these models, Thomas and Yoon-Nelson models well described the experimental data. Therefore, the result implies that nZVI-SS biocomposite is a cheap and most promising biosorbent for the removal of CV from wastewater.

Keywords: algae, biosorption, zero-valent, dye, wastewater

Procedia PDF Downloads 147

Authors: Dilesh Mogre, Jitendra Toravi, Saurabh Joshi, Prutha Deshpande, Aishwarya Kura

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In earlier days of technology, it was not possible to understand the nature of complex biomedical problems and were only left to clinical postulations. With advancement in science today, we have tools like Finite Element Modelling and simulation to solve complex biomedical problems. This paper presents how ANSYS WORKBENCH can be used to study deformation of pneumatized sphenoid bone caused by increased intracranial pressure. Intracranial pressure refers to the pressure inside the skull. The increase in the pressure above the normal range of 15mmhg can lead to serious conditions due to developed stresses and deformation. One of the areas where the deformation is suspected to occur is Sphenoid Bone. Moreover, the varying degree of pneumatization increases the complexity of the conditions. It is necessary to study deformation patterns on pneumatized sphenoid bone model at elevated intracranial pressure. Finite Element Analysis plays a major role in developing and analyzing model and give quantitative results.

Keywords: intracranial pressure, pneumatized sphenoid bone, deformation, finite element analysis

Procedia PDF Downloads 149

Authors: Hiroyuki Aoki

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The optoacoustic effect is the energy conversion phenomenon from light to sound. In recent years, this optoacoustic effect has been utilized for an imaging agent to visualize a tumor site in a living body. The optoacoustic imaging agent absorbs the light and emits the sound signal. The sound wave can propagate in a living organism with a small energy loss; therefore, the optoacoustic imaging method enables the molecular imaging of the deep inside of the body. In order to improve the imaging quality of the optoacoustic method, the more signal intensity is desired; however, it has been difficult to enhance the signal intensity of the optoacoustic imaging agent because the fundamental mechanism of the signal generation is unclear. This study deals with the mechanism to generate the sound wave signal from the optoacoustic imaging agent following the light absorption by experimental and theoretical approaches. The optoacoustic signal efficiency for the nano-particles consisting of metal and polymer were compared, and it was found that the polymer particle was better. The heat generation and transfer process for optoacoustic agents of metal and polymer were theoretically examined. It was found that heat generated in the metal particle rapidly transferred to the water medium, whereas the heat in the polymer particle was confined in itself. The confined heat in the small particle induces the massive volume expansion, resulting in the large optoacoustic signal for the polymeric particle agent. Thus, we showed that heat confinement is a crucial factor in designing the highly efficient optoacoustic imaging agent.

Keywords: nano-particle, opto-acoustic effect, in vivo imaging, molecular imaging

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Authors: Mario Sandro F. Rocha, Carlos S. Ande, Anderson A. Oliveira, Felipe M. Bersotti, Lucas O. Venzel

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The inertial motion capture systems (mocap) are among the most suitable tools for quantitative clinical analysis in rehabilitation and sports medicine. The inertial measuring units (IMUs), composed by accelerometers, gyroscopes, and magnetometers, are able to measure spatial orientations and calculate displacements with sufficient precision for applications in biomechanical analysis of movement. Furthermore, this type of system is relatively affordable and has the advantages of portability and independence from external references. In this work, we present the last version of our inertial motion capture system, based on the foregoing technology, with a unity interface designed for rehabilitation and sports. In our hardware architecture, only one serial port is required. First, the board client must be connected to the computer by a USB cable. Next, an available serial port is configured and opened to establish the communication between the client and the application, and then the client starts scanning for the active MOCAP_S servers around. The servers play the role of the inertial measuring units that capture the movements of the body and send the data to the client, which in turn create a package composed by the ID of the server, the current timestamp, and the motion capture data defined in the client pre-configuration of the capture session. In the current version, we can measure the game rotation vector (grv) and linear acceleration (lacc), and we also have a step detector that can be abled or disabled. The grv data are processed and directly linked to the bones of the 3D model, and, along with the data of lacc and step detector, they are also used to perform the calculations of displacements and other variables shown on the graphical user interface. Our user interface was designed to calculate and present variables that are important for rehabilitation and sports, such as cadence, speed, total gait cycle, gait cycle length, obliquity and rotation, and center of gravity displacement. Our goal is to present a low-cost portable and wearable system with a friendly interface for application in biomechanics and sports, which also performs as a product of high precision and low consumption of energy.

Keywords: biomechanics, inertial sensors, motion capture, rehabilitation

Procedia PDF Downloads 113

Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

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Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

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Authors: Ahmed Arkoazi, Hussein Znad, Ranjeet Utikar

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The synergistic impact and optimization of gas flow rate, concentration of CO₂, and light intensity on CO₂ biofixation rate were investigated using wastewater as a medium to cultivate Chlorella vulgaris under different conditions (gas flow rate 1-8 L/min), CO₂ concentration (0.03-7%), and light intensity (150-400 µmol/m².s)). Response Surface Methodology and Box-Behnken experimental Design were applied to find optimum values for gas flow rate, CO₂ concentration, and light intensity. The optimum values of the three independent variables (gas flow rate, concentration of CO₂, and light intensity) and desirability were 7.5 L/min, 3.5%, and 400 µmol/m².s, and 0.904, respectively. The highest amount of biomass produced and CO₂ biofixation rate at optimum conditions were 5.7 g/L, 1.23 gL^-1d^-1, respectively. The synergistic effect between gas flow rate and concentration of CO₂, and between gas flow rate and light intensity was significant on the three responses, while the effect between CO₂ concentration and light intensity was less significant on CO₂ biofixation rate. The results of this study could be highly helpful when using microalgae for CO₂ biofixation in wastewater treatment.

Keywords: bubble column reactor, gas holdup, hydrodynamics, sparger

Procedia PDF Downloads 110

Authors: Alessandro Falconieri, Sara De Vincentiis, Vittoria Raffa

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Mechanical force regulates axonal growth, elongation and maturation processes. This force is opening new frontiers in the field, contributing to a general understanding of the mechanisms of axon growth that, in the past, was thought to be governed exclusively by the growth cone and its ability to influence axonal growth in response to chemical signals. A method recently developed in our laboratory allows, through the labeling of neurons with magnetic nanoparticles (MNPs) and the use of permanent magnets, to apply extremely low mechanical forces, similar to those generated endogenously by the growth cone or by the increase of body mass during the organism growth. We found that these extremely low forces strongly enhance the spontaneous axonal elongation rate as well as neuronal sprouting. Data obtained don’t exclude that local phenomena, such as local transport and local translation, may be involved. These new advances could shed new light on what happens when the cell is subjected to external mechanical forces, opening new interesting scenarios in the field of mechanobiology.

Keywords: axon, external mechanical forces, magnetic nanoparticles, mechanotransduction

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Authors: Abdulhakeem Sulyman, Olukotun Zainab, Hammed Abdulquadri

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Lipases (triacylglycerol acyl hydrolases) (EC 3.1.1.3) are class of enzymes that catalyses the hydrolysis of triglycerides to glycerol and free fatty acids. They account for up to 10% of the enzyme in the market and have a wide range of applications in biofuel production, detergent formulation, leather processing and in food and feed processing industry. This research was conducted to study the effect of some metal ions on the activity of purified lipase produced by Aspergillus niger cultured on Vitellaria paradoxa shells. Purified lipase in 12.5 mM p-NPL was incubated with different metal ions (Zn²⁺, Ca²⁺, Mn²⁺, Fe²⁺, Na⁺, K⁺ and Mg²⁺). The final concentrations of metal ions investigated were 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1.0 mM. The results obtained from the study showed that Zn²⁺, Ca²⁺, Mn²⁺ and Fe²⁺ ions increased the activity of lipase up to 3.0, 3.0, 1.0, and 26.0 folds respectively. Lipase activity was partially inhibited by Na⁺ and Mg²⁺ with up to 88.5% and 83.7% loss of activity respectively. Lipase activity was also inhibited by K⁺ with up to 56.7% loss in the activity as compared to in the absence of metal ions. The study concluded that lipase produced by Aspergillus niger cultured on Vitellaria paradoxa shells can be activated by the presence of Zn²⁺, Ca²⁺, Mn²⁺ and Fe²⁺ and inhibited by Na⁺, K⁺ and Mg²⁺.

Keywords: Aspergillus niger, Vitellaria paradoxa, lipase, metal ions

Procedia PDF Downloads 124

Authors: Philip Semaha, Zhongfang Lei, Ziwen Zhao, Sen Liu, Zhenya Zhang, Kazuya Shimizu

Abstract:

The increasing generation of saline wastewater through various industrial activities is becoming a global concern for activated sludge (AS) based biological treatment which is widely applied in wastewater treatment plants (WWTPs). As for the AS process, an increase in wastewater salinity has negative impact on its overall performance. The advent of conventional aerobic granular sludge (AGS) or bacterial AGS biotechnology has gained much attention because of its superior performance. The development of algal-bacterial AGS could enhance better nutrients removal, potentially reduce aeration cost through symbiotic algae-bacterial activity, and thus, can also reduce overall treatment cost. Nonetheless, the potential of salt stress to decrease biomass growth, microbial activity and nutrient removal exist. Up to the present, little information is available on saline wastewater treatment by algal-bacterial AGS. To the authors’ best knowledge, a comparison of the two AGS systems has not been done to evaluate nutrients removal capacity in the context of salinity increase. This study sought to figure out the impact of salinity on the algal-bacterial AGS system in comparison to bacterial AGS one, contributing to the application of AGS technology in the real world of saline wastewater treatment. In this study, the salt concentrations tested were 0 g/L, 1 g/L, 5 g/L, 10 g/L and 15 g/L of NaCl with 24-hr artificial illuminance of approximately 97.2 µmol m¯²s¯¹, and mature bacterial and algal-bacterial AGS were used for the operation of two identical sequencing batch reactors (SBRs) with a working volume of 0.9 L each, respectively. The results showed that salinity increase caused no apparent change in the color of bacterial AGS; while for algal-bacterial AGS, its color was progressively changed from green to dark green. A consequent increase in granule diameter and fluffiness was observed in the bacterial AGS reactor with the increase of salinity in comparison to a decrease in algal-bacterial AGS diameter. However, nitrite accumulation peaked from 1.0 mg/L and 0.4 mg/L at 1 g/L NaCl in the bacterial and algal-bacterial AGS systems, respectively to 9.8 mg/L in both systems when NaCl concentration varied from 5 g/L to 15 g/L. Almost no ammonia nitrogen was detected in the effluent except at 10 g/L NaCl concentration, where it averaged 4.2 mg/L and 2.4 mg/L, respectively, in the bacterial and algal-bacterial AGS systems. Nutrients removal in the algal-bacterial system was relatively higher than the bacterial AGS in terms of nitrogen and phosphorus removals. Nonetheless, the nutrient removal rate was almost 50% or lower. Results show that algal-bacterial AGS is more adaptable to salinity increase and could be more suitable for saline wastewater treatment. Optimization of operation conditions for algal-bacterial AGS system would be important to ensure its stably high efficiency in practice.

Keywords: algal-bacterial aerobic granular sludge, bacterial aerobic granular sludge, Nutrients removal, saline wastewater, sequencing batch reactor

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Authors: Meenakshi Srivastava, A. K. Mishra

Abstract:

This study characterizes the metabolic versatility of denitrifying bacterial communities residing in the paddy soil using the GC-MS based Phospholipid Fatty Acid (PLFA) analyses simultaneously with nosZ gene based PCR-DGGE (Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis) and real time Q-PCR analysis. We have analyzed the abundance of nitrous oxide reductase (nosZ) genes, which was subsequently related to soil PLFA profile and DGGE based denitrifier community structure. Soil denitrifying bacterial community comprised majority or dominance of Ochrobactrum sp. following Cupriavidus and uncultured bacteria strains in paddy soil of selected sites. Initially, we have analyzed the abundance of the nitrous oxide reductase gene (nosZ), which was found to be related with PLFA based lipid profile. Chandauli of Eastern UP, India represented greater amount of lipid content (C18-C20) and denitrifier’s diversity. This study suggests the positive co-relation between soil PLFA profiles, DGGE, and Q-PCR data. Thus, a close networking among metabolic abilities and taxonomic composition of soil microbial communities existed, and subsequently, such work at greater extent could be helpful in managing nutrient dynamics as well as microbial dynamics of paddy soil ecosystem.

Keywords: denaturing gradient gel electrophoresis, DGGE, nitrifying and denitrifying bacteria, PLFA, Q-PCR

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Authors: Shazia Bano

Abstract:

Nanoconjugates that integrate photo-based therapeutics and diagnostics within a single platform promise great advances in revolutionizing cancer treatments. However, to achieve high therapeutic efficacy, designing functionally efficacious nanocarriers to tightly retain the drug, promoting selective drug localization and release, and the validation of the efficacy of these nanoconjugates is a great challenge. Here we have designed smart multiagent, liposome based targeted photoactivatable multiagent nanoconjugates, doped with a photoactivatable chromophore benzoporphyrin derivative (BPD) labelled with an active targeting ligand cetuximab to target the EGFR receptor (over expressed in various cancer cells) to deliver a combination of therapeutic agents. This study establishes a tunable nanoplatform for the delivery of the photoactivatable multiagent nanoconjugates for tumor-specific accumulation and targeted destruction of cancer cells in complex cancer model to enhance the therapeutic index of the administrated drugs.

Keywords: targeting, photodynamic therapy, photoactivatable, nanoconjugates

Procedia PDF Downloads 115

Authors: Lukasz Szydlowski, Jiri Ehlich, Igor Goryanin

Abstract:

We demonstrate a single chamber, 96-well-plated based Microbial Fuel Cell (MFC) with printed, electronic components. This invention is aimed at robust selection of electrogenic microbial community under specific conditions, e.g., electrode potential, pH, nutrient concentration, salt concentration that can be altered within the 96 well plate array. This invention enables robust selection of electrogenic microbial community under the homogeneous reactor, with multiple conditions that can be altered to allow comparative analysis. It can be used as a standalone technique or in conjunction with other selective processes, e.g., flow cytometry, microfluidic-based dielectrophoretic trapping. Mobile conductive elements, like carbon paper, carbon sponge, activated charcoal granules, metal mesh, can be inserted inside to increase the anode surface area in order to collect electrogenic microorganisms and to transfer them into new reactors or for other analytical works. An array of 96-well plate allows this device to be operated by automated pipetting stations.

Keywords: bioengineering, electrochemistry, electromicrobiology, microbial fuel cell

Procedia PDF Downloads 114

Authors: Thi Thao Ninh, Yuri Trusov, José Ramón Botella

Abstract:

Heterotrimeric G proteins, comprised of three subunits, α, β and γ, are involved in signal transduction pathways that mediate a vast number of processes across the eukaryotic kingdom. 23 Gα subunits are present in humans whereas most plant genomes encode for only one canonical Gα. The disparity observed between Arabidopsis, rice, and maize Gα-deficient mutant phenotypes suggest that Gα functions have diversified between eudicots and monocots during evolution. Alternatively, since the only Gα mutations available in dicots have been produced in Arabidopsis, the possibility exists that this species might be an exception to the rule. In order to test this hypothesis, we studied the G protein α subunit (TGA1) in tomato. Four tga1 knockout lines were generated in tomato cultivar Moneymaker using CRISPR/Cas9. The tga1 mutants exhibit a number of auxin-related phenotypes including changes in leaf shape, reduced plant height, fruit size and number of seeds per fruit. In addition, tga1 mutants have increased sensitivity to abscisic acid during seed germination, reduced sensitivity to exogenous auxin during adventitious root formation from cotyledons and excised hypocotyl explants. Our results suggest that Gα mutant phenotypes in tomato are very similar to those observed in monocots, i.e. rice and maize, and cast doubts about the validity of using Arabidopsis as a model system for plant G protein studies.

Keywords: auxin-related phenotypes, CRISPR/Cas9, G protein α subunit, heterotrimeric G proteins, tomato

Procedia PDF Downloads 102

Authors: Bao-Fang Wen, Meng-Na Dai, Gao-Pei Zhu, Chen-Xi Zhang, Jing Sun, Xun-Bao Yin, Yu-Han Zhao, Hong-Wei Sun, Wei-Fen Zhang

Abstract:

A drug-loaded nanoparticles containing berberine hydrochloride (BH/FA-CTS-NPs) was prepared. The physicochemical characterizations of BH/FA-CTS-NPs and the inhibitory effect on the HeLa cells were investigated. Folic acid-conjugated chitosan (FA-CTS) was prepared by amino reaction of folic acid active ester and chitosan molecules; BH/FA-CTS-NPs were prepared using ionic cross-linking technique with BH as a model drug. The morphology and particle size were determined by Transmission Electron Microscope (TEM). The average diameters and polydispersity index (PDI) were evaluated by Dynamic Light Scattering (DLS). The interaction between various components and the nanocomplex were characterized by Fourier Transform Infrared Spectroscopy (FT-IR). The entrapment efficiency (EE), drug-loading (DL) and in vitro release were studied by UV spectrophotometer. The effect of cell anti-migratory and anti-invasive actions of BH/FA-CTS-NPs were investigated using MTT assays, wound healing assays, Annexin-V-FITC single staining assays, and flow cytometry, respectively. HeLa nude mice subcutaneously transplanted tumor model was established and treated with different drugs to observe the effect of BH/FA-CTS-NPs in vivo on HeLa bearing tumor. The BH/FA-CTS-NPs prepared in this experiment have a regular shape, uniform particle size, and no aggregation phenomenon. The results of DLS showed that mean particle size, PDI and Zeta potential of BH/FA-CTS NPs were (249.2 ± 3.6) nm, 0.129 ± 0.09, 33.6 ± 2.09, respectively, and the average diameter and PDI were stable in 90 days. The results of FT-IR demonstrated that the characteristic peaks of FA-CTS and BH/FA-CTS-NPs confirmed that FA-CTS cross-linked successfully and BH was encapsulated in NPs. The EE and DL amount were (79.3 ± 3.12) % and (7.24 ± 1.41) %, respectively. The results of in vitro release study indicated that the cumulative release of BH/FA-CTS NPs was (89.48±2.81) % in phosphate-buffered saline (PBS, pH 7.4) within 48h; these results by MTT assays and wund healing assays indicated that BH/FA-CTS NPs not only inhibited the proliferation of HeLa cells in a concentration and time-dependent manner but can induce apoptosis as well. The subcutaneous xenograft tumor formation rate of human cervical cancer cell line HeLa in nude mice was 98% after inoculation for 2 weeks. Compared with BH group and BH/CTS-NPs group, the xenograft tumor growth of BH/FA-CTS-NPs group was obviously slower; the result indicated that BH/FA-CTS-NPs could significantly inhibit the growth of HeLa xenograft tumor. BH/FA-CTS NPs with the sustained release effect could be prepared successfully by the ionic crosslinking method. Considering these properties, block proliferation and impairing the migration of the HeLa cell line, BH/FA-CTS NPs could be an important compound for consideration in the treatment of cervical cancer.

Keywords: folic-acid, chitosan, berberine hydrochloride, nanoparticles, cervical cancer

Procedia PDF Downloads 92

Authors: Claudia Matassa, Matthias Hohne, Dominic Ormerod, Yamini Satyawali

Abstract:

Chiral amines integrate the backbone of several active pharmaceutical ingredients (APIs) used in modern medicine for the treatment of a vast range of diseases. Despite the demand, their synthesis remains challenging. Besides a range of chemicals and enzymatical methods, chiral amine synthesis using transaminases (EC 2.6.1.W) represents a useful alternative to access this important class of compounds. Even though transaminases exhibit excellent stereo and regioselectivity and the potential for high yield, the reaction suffers from a number of challenges, including the thermodynamic equilibrium, product inhibition, and low substrate solubility. In this work, we demonstrate a membrane assisted strategy for addressing these challenges. It involves the use of high molecular weight (HMW) amine donors for the transaminase-catalyzed synthesis of 4-phenyl-2-butylamine in both aqueous and organic solvent media. In contrast to common amine donors such as alanine or isopropylamine, these large molecules, provided in excess for thermodynamic equilibrium shifting, are easily retained by commercial nanofiltration membranes; thus a selective permeation of the desired smaller product amine is possible. The enzymatic transamination in aqueous media, combined with selective product removal shifted the equilibrium enhancing substrate conversion by an additional 25% compared to the control reaction. Along with very efficient amine product removal, there was undesirable loss of ketone substrate and low product concentration was achieved. The system was therefore further improved by performing the reaction in organic solvent (n-heptane). Coupling the reaction system with membrane-assisted product removal resulted in a highly concentrated and relatively pure ( > 97%) product solution. Moreover, a product yield of 60% was reached, compared to 15% without product removal.

Keywords: amine donor, chiral amines, in situ product removal, transamination

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Authors: Xiao L Dai, Wen X Wei, Shuo Wang, Jia B Li, Yan Wei

Abstract:

Heavy oil pollution generated from both natural and anthropogenic sources could cause significant damages to the ecological environment, due to the toxicity of some of its constituents. Nowadays, microbial remediation is becoming a promising technology to treat oil pollution owing to its low cost and prevention of secondary pollution; microorganisms are key players in the process. Compared to the free microorganisms, immobilized microorganisms possess several advantages, including high metabolic activity rates, strong resistance to toxic chemicals and natural competition with the indigenous microorganisms, and effective resistance to washing away (in open water system). Many immobilized microorganisms have been successfully used for bioremediation of heavy oil pollution. Considering the broad choices, low cost, simple process, large specific surface area and less impact on microbial activity, modified zeolite were selected as a bio-carrier for bacteria immobilization. Three strains of heavy oil-degrading bacteria Bacillus sp. DL-13, Brevibacillus sp. DL-1 and Acinetobacter sp. DL-34 were immobilized on the modified zeolite under mild conditions, and the bacterial load (bacteria /modified zeolite) was 1.12 mg/g, 1.11 mg/g, and 1.13 mg/g, respectively. SEM results showed that the bacteria mainly adsorbed on the surface or punctured in the void of modified zeolite. The heavy oil degradation efficiency of immobilized bacteria was 62.96%, higher than that of the free bacteria (59.83%). The heavy oil degradation process of immobilized bacteria accords with the first-order reaction equation, and the reaction rate constant is 0.1483 d⁻¹, which was significantly higher than the free bacteria (0.1123 d⁻¹), suggesting that the immobilized bacteria can rapidly start up the heavy oil degradation and has a high activity of heavy oil degradation. The results suggested that immobilized bacteria are promising technology for bioremediation of oil pollution.

Keywords: heavy oil pollution, microbial remediation, modified zeolite, immobilized bacteria

Procedia PDF Downloads 113

Authors: Ketaki D. Belsare, Nicholas F. Polizzi, Lior Shtayer, William F. DeGrado

Abstract:

Nature utilizes metalloproteins to perform chemical transformations with activities and selectivities that have long been the inspiration for design principles in synthetic and biological systems. The chemical reactivities of metalloproteins are directly linked to local environment effects produced by the protein matrix around the metal cofactor. A complete understanding of how the protein matrix provides these interactions would allow for the design of functional metalloproteins. The de novo computational design of proteins have been successfully used in design of active sites that bind metals like di-iron, zinc, copper containing cofactors; however, precisely designing active sites that can bind small molecule ligands (e.g., substrates) along with metal cofactors is still a challenge in the field. The de novo computational design of a functional metalloprotein that contains a purposefully designed substrate binding site would allow for precise control of chemical function and reactivity. Our research strategy seeks to elucidate the design features necessary to bind the cofactor protoporphyrin IX (hemin) in close proximity to a substrate binding pocket in a four helix bundle. First- and second-shell interactions are computationally designed to control orientation, electronic structure, and reaction pathway of the cofactor and substrate. The design began with a parameterized helical backbone that positioned a single histidine residue (as an axial ligand) to receive a second-shell H-bond from a Threonine on the neighboring helix. The metallo-cofactor, hemin was then manually placed in the binding site. A structural feature, pi-bulge was introduced to give substrate access to the protoporphyrin IX. These de novo metalloproteins are currently being tested for their activity towards hydroxylation and epoxidation. The de novo designed protein shows hydroxylation of aniline to 4-aminophenol. This study will help provide structural information of utmost importance in understanding de novo computational design variables impacting the functional activities of a protein.

Keywords: metalloproteins, protein design, de novo protein, biocatalysis

Procedia PDF Downloads 127