Search results for: 35 kDa hyaluronan fragment HA35
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 175

Search results for: 35 kDa hyaluronan fragment HA35

85 Genetic Characterization of Acanthamoeba Isolates from Amoebic Keratitis Patients

Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

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Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

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84 Efficiency of PCR-RFLP for the Identification of Adulteries in Meat Formulation

Authors: Hela Gargouri, Nizar Moalla, Hassen Hadj Kacem

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Meat adulteration affecting the safety and quality of food is becoming one of the main concerns of public interest across the world. The drastic consequences on the meat industry highlighted the urgent necessity to control the products' quality and to point out the complexity of both supply and processing circuits. Due to the expansion of this problem, the authentic testing of foods, particularly meat and its products, is deemed crucial to avoid unfair market competition and to protect consumers from fraudulent practices of meat adulteration. The adoption of authentication methods by the food quality-control laboratories is becoming a priority issue. However, in some developing countries, the number of food tests is still insignificant, although a variety of processed and traditional meat products are widely consumed. Little attention has been paid to provide an easy, fast, reproducible, and low-cost molecular test, which could be conducted in a basic laboratory. In the current study, the 359 bp fragment of the cytochrome-b gene was mapped by PCR-RFLP using firstly fresh biological supports (DNA and meat) and then turkey salami as an example of commercial processed meat. This technique has been established through several optimizations, namely: the selection of restriction enzymes. The digestion with BsmAI, SspI, and TaaI succeed to identify the seven included animal species when meat is formed by individual species and when the meat is a mixture of different origin. In this study, the PCR-RFLP technique using universal primer succeed to meet our needs by providing an indirect sequencing method identifying by restriction enzymes the specificities characterizing different species on the same amplicon reducing the number of potential tests.

Keywords: adulteration, animal species, authentication, meat, mtDNA, PCR-RFLP

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83 Anatomical and Histological Characters of Cymbopogon nardus Roots and Its Mutagenic Properties

Authors: Pravaree Phuneerub, Chanida Palanuvej, Nijsiri Ruangrungsi

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Cymbopogon nardus Rendel (Family Gramineae) is commonly known as citronella grass. The dried root of C. nardus is used for antipyretic, anti-inflammation, anti-analgesic and anticancer in traditional Thai medicine. Transverse sectional and pulverized C. nardus root were illustrated. The volatile oil was extracted from oil gland by hydrodistillation and analysed by GC/MS. Cymbopogon nardus root was exhaustively extracted by continuously maceration in ethanol and water respectively. The mutagenic and antimutagenic properties of the ethanol extract and fractionated water extract of C. nardus root were evaluated by Ames assay using the S. typhimurium strains TA98 and TA100 as the models. The result indicated that the anatomical character of root transverse section displayed epidermis, parenchyma, oil gland, phloem, xylem vessel, endodermis and pith. Histological characters of root powder showed parenchyma containing oleoresin, parenchyma in longitudinal view, reticulate vessel, annular vessel, starch granules and fragment of fiber. The root volatile oil was rich in sesquiterpenes dominated by elemol (22.87%) and alpha-eudesmol (16.09%). For mutagenic activity, the both extracts of C. nardus were no mutagenic toward S. typhimurium strains TA98 and TA100. Furthermore, the ethanol extract and fractionated water extract of C. nardus root demonstrated strong antimutagenic effect against of nitrite treated 1-aminopyrene to S. typhimurium strains TA98 and TA100. This present investigation suggested that the dried root extract of C. nardus can be further developed as promising antimutagenic agent.

Keywords: Cymbopogon nardus, volatile oil analysis, mutagenic, antimutagenic effect, Ames Salmonella assay

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82 Genetic and Phenotypic Variability Among the Vibrio Cholerae O1 Isolates of India

Authors: Sreeja Shaw, Prosenjit Samanta, Asish Kumar Mukhopadhyay

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Cholera is still a global public health burden and is caused by Vibrio cholerae O1 and O139 serogroups. Evidence from recent outbreaks in Haiti and Yemen suggested that circulating V. cholerae O1 El Tor variant strains are continuously changing to cause more ruinous outbreaks worldwide, and most of them have emerged from the Indian subcontinents. Therefore, we studied the changing virulence characteristics along with the antibiotic resistance profile of V. cholerae O1strains isolated from seasonal outbreaks in three cholera endemic regions during 2018, Gujarat and Maharashtra in Western India (87 strains), and to compare those features with the isolates of West Bengal in Eastern India (48 strains) collected during the same period. All the strains from Western India were of Ogawa serotype, polymyxin B-sensitive, hemolytic, and contained a large fragment deletion in VSP-II genomic region similar with Yemen outbreak strains and carried more virulent Haitian genetic alleles of major virulence associated genes ctxB, tcpA, and rtxA. Conversely, 14.6% (7/48) of the strains from Eastern India were belong to the Inaba serotype, polymyxin B-resistant, non-hemolytic, harbored intact VSP-II region, classical ctxB, Haitian tcpA, and El Tor rtxA alleles. Interestingly, resistance to tetracycline and chloramphenicol was seen in isolates from both regions, which are not very common among V. cholerae O1 isolates in India. Therefore, this study indicated West Bengal as a diverse region where two different types of El Tor variant hypervirulent strains are co-existed, probably competing for their better environmental survival, which may result in severe irrepressible disease outcome in the future.

Keywords: cholera, vibrio cholerae, polymyxin B, Non-hemolytic, ctxB, tcpA, rtxA, VSP-II

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81 Genetic Diversity of Tiger Groupers (Epinephelus fuscoguttatus) Challenged with Vibrio Parahaemolyticus and Exposed to Extreme Low Salinities

Authors: Hidayah Triana, Mahir S. Gani, Asmi Citra Malina, Hamka

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This study was conducted to determine genetic diversity of tiger groupers that are resistant to V. parahaemolyticus and tolerant to low extreme salinities. This research is useful to obtain superior broodstock of fish. Tiger grouper used were 6 to 8 cm obtained from Brackish Water Aquaculture Research Center Gondol (Bali). This study consists of four stages: preliminary stage was adaptation of fish exposed to several concentrations of V. parahaemolyticus (103, 104, 105, 106, and 107 CFU / ml); second stage was test of Lethal Concentration (LC50) of bacteria to fish; third stage was salinity tolerance test (low salinity 12, 14 and 16 ppt) and fourth stage was analysis of DNA profiles. For DNA profiles analysis, genomic DNA of fish were extracted for PCR using primers YNZ-22 and UBC-122 and visualized by electrophoresis method. The results showed that Lethal concentration of bacteria (LC50) to fish was 1,56x106 CFU/ml. Furthermore, survival rate of groupers exposed with low salinities (12, 14, 16 ppt) survival rates were found to be 54,17 %, 66,67 % and 79,16 % respectively. Average of DNA fragment (5 fragments) generated from primer UBC-122 in the group of fish resistant to V.parahaemolyticus and tolerant to low salinities was similar to group of susceptible to low salinities. Primer YNZ-22 generated more diverse of DNA fragments (8,0 and 5,8 fragments) both in the group of fish tolerant and susceptible to low salinities compared to primer UBC-122 (5,0 fragments). Size of DNA 1.5 kb resulted from primer YNZ-22. Primer YNZ-22 generated 4 (50 %) and 3 (42,8 %) polymorfic fragments in the group of fish tolerant and susceptible to low salinities, respectively. Four (4) monomorfic fragments were found both in the group of fish tolerant and susceptible to low salinities. Primer UBC-122 generated 6 (85,7 %) and 9 (90,0 %) polymorfic fragments in the fish tolerant and susceptible to low salinities, respectively.

Keywords: genetic diversity, epinephelus fuscoguttatus, V. parahaemolyticus, PCR-RAPD, low extreme salinity

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80 Texture Characterization and Mineralogical Composition of the 1982-1983 Second Phase Galunggung Eruption, West Java Regency, Indonesia

Authors: M. Hanif Irsyada, Rifaldy, Arif Lutfi Namury, Syahreza S. Angkasa, Khalid Rizky, Ricky Aryanto, M. Alfiyan Bagus, Excobar Arman, Fahri Septianto, Firman Najib Wibisana

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Galunggung Mountain is an active volcano in Indonesia, precisely on the island of Java. This area is included in the Sunda Sunda arc formed by the tendency of the Australian oceanic plate to Eurasian continental plate. This research was conducted to determine the characteristics and document the mineralogical composition of the Galunggung eruption of the second phase 1982-1983. In fragment samples, petrographic analysis is carried out under a qualitative and quantitative polarizing microscope. This sample was obtained from the second phase eruption in the Cibanjanj formation. Based on the analysis results obtained filter texture characteristics, olivine parallel growth, lamellar structure, glass inclusion, plagioclase zonation and obtained special texture in the gabbroic cummulate. The mineral composition consists of phenocryst plagioclase (41vol%), pyroxene (26vol%), olivin (4vol%) and mineral opaque (29vol%). Microlite minerals consist of plagioclase (31.95vol%), pyroxene (12.09vol%), opaque minerals (55.96vol%). This research is expected to be developed by further researchers to be able to explain in more detail related to Galunggung mountain with 3 phases of eruption that are so intense. Also, it is expected to explain the structural characteristics and mineralogical composition that can be used to determine the origin of all the results of the Galunggung eruption 1982-1983.

Keywords: Galunggung eruption, mineralogical composition, texture characterization, gabbroic cumulate

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79 Voters' Acceptance of Anti-guardians' Narratives: Electoral Politics in Establishmentarian Democracies

Authors: Rai Mansoor Imtiaz

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Guardians in hybrid regimes fragment opposition parties and ban their political leaders, and disenfranchise their voters' political participation. When guardians in hybrid regimes are so powerful that they remain decisive on electoral politics of states, and have powers to ban political parties and their leadership, then "why do political parties backed by those powerful guardians lose elections" and "how do anti-establishment parties make electoral inroads at the local and national levels." These two questions are interrelated with the key research question of my research "why do people vote for political parties rejected by powerful guardians in establishmentarian democracies." Furthermore, this research question is important to be explored for two reasons. First, existing literature only reflects the electoral victories of opposition parties or defeats of military-sponsored parties (see Thailand and Turkey) but remains silent on political change that led the anti-military parties to win the elections. Second, why is it a case that people belonging to the countries where militaries remain popular among the public (see Turkey and Pakistan) have started putting their trust in anti-establishment politicians who criticise the military against their intervention in politics? For instance, in Pakistan, where commenting against the military is meant to comment against the state –– an anti-military narrative is getting popular support. The conceptual framework of hybrid states in this research relies on the concept of a 'reserved domain/tutelary body' (guardians of hybrid states). However, this research makes a case that hybrid states are not consolidated separate political entities but rather vacillated states that fluctuate between democratic and authoritarian practices. This paper, therefore, uses the term establishmentarian democracy as a subtype of the hybrid regime, which is more consolidated than a hybrid democracy.

Keywords: Guardians, Hybrid Regimes, Voters, Elections, Democracy, South Asia

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78 Effect of Fiddler Crab Burrows on Bacterial Communities of Mangrove Sediments

Authors: Mohammad Mokhtari, Gires Usup, Zaidi Che Cob

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Bacteria communities as mediators of the biogeochemical process are the main component of the mangrove ecosystems. Crab burrows by increasing oxic-anoxic interfaces and facilitating the flux rate between sediment and tidal water affect biogeochemical properties of sediments. The effect of fiddler crab burrows on the density and diversity of bacteria were investigated to elucidate the effect of burrow on bacterial distribution. Samples collected from the burrow walls of three species of fiddler crabs including Uca paradussumieri, Uca rosea, and Uca forcipata. Sediment properties including grain size, temperature, Redox potential, pH, chlorophyll, water and organic content were measured from the burrow walls to assess the correlation between environmental variables and bacterial communities. Bacteria were enumerated with epifluorescence microscopy after staining with SYBR green. Bacterial DNA extracted from sediment samples and the community profiles of bacteria were determined with Terminal Restriction Fragment Length Polymorphism (T-RFLP). High endemism was observed among bacterial communities. Among the 152 observed OTU’s, 22 were found only in crab burrows. The highest bacterial density and diversity were recorded in burrow wall. The results of ANOSIM indicated a significant difference between the bacterial communities from the three species of fiddler crab burrows. Only 3% of explained bacteria variability in the constrained ordination model of CCA was contributed to depth, while much of the bacteria’s variability was attributed to coarse sand, pH, and chlorophyll content. Our findings suggest that crab burrows by affecting sediment properties such as redox potential, pH, water, and chlorophyll content induce significant effects on the bacterial communities.

Keywords: bioturbation, canonical corresponding analysis, fiddler crab, microbial ecology

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77 Prediction of Ionizing Radiation Doses in Irradiated red Pepper (Capsicum annuum) and Mint (Mentha piperita) by Gel Electrophoresis

Authors: Şeyma Özçirak Ergün, Ergün Şakalar, Emrah Yalazi̇, Nebahat Şahi̇n

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Food irradiation is a usage of exposing food to ionising radiation (IR) such as gamma rays. IR has been used to decrease the number of harmful microorganisms in the food such as spices. Excessive usage of IR can cause damage to both food and people who consuming food. And also it causes to damages on food DNA. Generally, IR detection techniques were utilized in literature for spices are Electron Spin Resonance (ESR), Thermos Luminescence (TL). Storage creates negative effect on IR detection method then analyses of samples have been performed without storage in general. In the experimental part, red pepper (Capsicum annuum) and mint (Mentha piperita) as spices were exposed to 0, 0.272, 0.497, 1.06, 3.64, 8.82, and 17.42 kGy ionize radiation. ESR was applied to samples irradiated. DNA isolation from irradiated samples was performed using GIDAGEN Multi Fast DNA isolation kit. The DNA concentration was measured using a microplate reader spectrophotometer (Infinite® 200 PRO-Life Science–Tecan). The concentration of each DNA was adjusted to 50 ng/µL. Genomic DNA was imaged by UV transilluminator (Gel Doc XR System, Bio-Rad) for the estimation of genomic DNA bp-fragment size after IR. Thus, agarose gel profiles of irradiated spices were obtained to determine the change of band profiles. Besides, samples were examined at three different time periods (0, 3, 6 months storage) to show the feasibility of developed method. Results of gel electrophoresis showed especially degradation of DNA of irradiated samples. In conclusion, this study with gel electrophoresis can be used as a basis for the identification of the dose of irradiation by looking at degradation profiles at specific amounts of irradiation. Agarose gel results of irradiated samples were confirmed with ESR analysis. This method can be applied widely to not only food products but also all biological materials containing DNA to predict radiation-induced damage of DNA.

Keywords: DNA, electrophoresis, gel electrophoresis, ionizeradiation

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76 Conserving History: Evaluating and Selecting Effective Restoration Methods for a Fragment Mural Painting from Amarna

Authors: Kholod Khairy Salama, Shabban Hassan Thabet

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In the present study, a comprehensive investigation has been undertaken into an Egyptian mural painting with feet wear slippers approach to choose the most successful restoration methods. The mural painting under examination dates back to the Amarna period; it was detached from a wall of an unknown tomb in Egypt, and currently, it is initially displayed in a showcase at the Egyptian Museum – Tahrir Square – Cairo, Egypt. The main objectives of this research were to (a) reveal the pigment used in the mural painting, (b) reveal the medium used with colours, (c) determine the technique of manufacturing, (e) determine the ground support, and (f) reveal the main deterioration aspects. The analytical techniques used for investigation were Optical Microscopy, Raman, X-ray Florescence, X-ray diffraction, and Fourier transform infrared coupled with attenuated total reflectance “FTIR-ATR”. The investigation revealed that the vital deterioration factors affecting the object. This research aims to examine and analyze the mural painting to choose the suitable method for the restoration process (a) define the colours through comparative analysis to choose the suitable material for cleaning, (b) define the natural structure of the ground support layer, which appeared as mud layer (c) determine the medium used with colours (d) diagnosis the presence of the white wash layer, and (e) choose the suitable restoration methods according to the results. Conclusion: This study focused mainly on the physical and chemical properties of the mural painting compound and the main changes that happened to the mural painting material, which caused deterioration and fall down of the painting parts, so we can find the best and optimum restoration ways for this object.

Keywords: mural paintings, Tal Al-Amarna, digital microscope, Raman, XRF, XRD, FTIR

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75 Optimization for Guide RNA and CRISPR/Cas9 System Nanoparticle Mediated Delivery into Plant Cell for Genome Editing

Authors: Andrey V. Khromov, Antonida V. Makhotenko, Ekaterina A. Snigir, Svetlana S. Makarova, Natalia O. Kalinina, Valentin V. Makarov, Mikhail E. Taliansky

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Due to its simplicity, CRISPR/Cas9 has become widely used and capable of inducing mutations in the genes of organisms of various kingdoms. The aim of this work was to develop applications for the efficient modification of DNA coding sequences of phytoene desaturase (PDS), coilin and vacuolar invertase (Solanum tuberosum) genes, and to develop a new nanoparticles carrier efficient technology to deliver the CRISPR/Cas9 system for editing the plant genome. For each of the genes - coilin, PDS and vacuolar invertase, five single RNA guide (sgRNAs) were synthesized. To determine the most suitable nanoplatform, two types of NP platforms were used: magnetic NPs (MNPS) and gold NPs (AuNPs). To test the penetration efficiency, they were functionalized with fluorescent agents - BSA * FITS and GFP, as well as labeled Cy3 small-sized RNA. To measure the efficiency, a fluorescence and confocal microscopy were used. It was shown that the best of these options were AuNP - both in the case of proteins and in the case of RNA. The next step was to check the possibility of delivering components of the CRISPR/Cas9 system to plant cells for editing target genes. AuNPs were functionalized with a ribonucleoprotein complex consisting of Cas9 and corresponding to target genes sgRNAs, and they were biolistically bombarded to axillary buds and apical meristems of potato plants. After the treatment by the best NP carrier, potato meristems were grown to adult plants. DNA isolated from this plants was sent to a preliminary fragment of the analysis to screen out the non-transformed samples, and then to the NGS. The present work was carried out with the financial support from the Russian Science Foundation (grant No. 16-16-04019).

Keywords: biobombardment, coilin, CRISPR/Cas9, nanoparticles, NPs, PDS, sgRNA, vacuolar invertase

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74 NeuroBactrus, a Novel, Highly Effective, and Environmentally Friendly Recombinant Baculovirus Insecticide

Authors: Yeon Ho Je

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A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties, such as a higher insecticidal activity and improved recovery, compared to wild-type baculovirus. For the construction of NeuroBactrus, the Bacillus thuringiensis crystal protein gene (here termed cry1-5) was introduced into the Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of the polyhedrin–cry1-5–polyhedrin genes under the control of the polyhedrin promoter. In the opposite direction, an insect-specific neurotoxin gene, AaIT, from Androctonus australis was introduced under the control of an early promoter from Cotesia plutellae bracovirus by fusion of a partial fragment of orf603. The polyhedrin–Cry1-5–polyhedrin fusion protein expressed by the NeuroBactrus was not only occluded into the polyhedra, but it was also activated by treatment with trypsin, resulting in an_65-kDa active toxin. In addition, quantitative PCR revealed that the neurotoxin was expressed from the early phase of infection. NeuroBactrus showed a high level of insecticidal activity against Plutella xylostella larvae and a significant reduction in the median lethal time against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Rerecombinant mutants derived from NeuroBactrus in which AaIT and/or cry1-5 were deleted were generated by serial passages in vitro. Expression of the foreign proteins (B. thuringiensis toxin and AaIT) was continuously reduced during the serial passage of the NeuroBactrus. Moreover, polyhedra collected from S. exigua larvae infected with the serially passaged NeuroBactrus showed insecticidal activity similar to that of wild-type AcMNPV. These results suggested that NeuroBactrus could be recovered to wild-type AcMNPV through serial passaging.

Keywords: baculovirus, insecticide, neurotoxin, neurobactrus

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73 Biosafety Study of Genetically Modified CEMB Sugarcane on Animals for Glyphosate Tolerance

Authors: Aminah Salim, Idrees Ahmed Nasir, Abdul Qayyum Rao, Muhammad Ali, Muhammad Sohail Anjum, Ayesha Hameed, Bushra Tabassum, Anwar Khan, Arfan Ali, Mariyam Zameer, Tayyab Husnain

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Risk assessment of transgenic herbicide tolerant sugarcane having CEMB codon optimized cp4EPSPS gene was done in present study. Fifteen days old chicks taken from K&Ns Company were randomly assorted into four groups with eight chicks in each group namely control chicken group fed with commercial diet, non-transgenic group fed with non-experimental sugarcane and transgenic group fed with transgenic sugarcane with minimum and maximum level. Body weights, biochemical analysis for Urea, alkaline phosphatase, alanine transferase, aspartate transferase, creatinine and bilirubin determination and histological examination of chicks fed with four types of feed was taken at fifteen days interval and no significant difference was observed in body weight biochemical and histological studies of all four groups. Protein isolated from the serum sample was analyzed through dipstick and SDS-PAGE, showing the absence of transgene protein in the serum sample of control and experimental groups. Moreover the amplification of cp4EPSPS gene with gene specific primers of DNA isolated from chicks blood and also from commercial diet was done to determine the presence and mobility of any nucleotide fragment of the transgene in/from feed and no amplification was obtained in feed as well as in blood extracted DNA of any group. Also no mRNA expression of cp4EPSPS gene was obtained in any tissue of four groups of chicks. From the results it is clear that there is no deleterious or harmful effect of the CEMB codon optimized transgenic cp4EPSPS sugarcane on the chicks health.

Keywords: chicks, cp4EPSPS, glyphosate, sugarcane

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72 Development of Immuno-Modulators: Application of Molecular Dynamics Simulation

Authors: Ruqaiya Khalil, Saman Usmani, Zaheer Ul-Haq

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The accurate characterization of ligand binding affinity is indispensable for designing molecules with optimized binding affinity. Computational tools help in many directions to predict quantitative correlations between protein-ligand structure and their binding affinities. Molecular dynamics (MD) simulation is a modern state-of-the-art technique to evaluate the underlying basis of ligand-protein interactions by characterizing dynamic and energetic properties during the event. Autoimmune diseases arise from an abnormal immune response of the body against own tissues. The current regimen for the described condition is limited to immune-modulators having compromised pharmacodynamics and pharmacokinetics profiles. One of the key player mediating immunity and tolerance, thus invoking autoimmunity is Interleukin-2; a cytokine influencing the growth of T cells. Molecular dynamics simulation techniques are applied to seek insight into the inhibitory mechanisms of newly synthesized compounds that manifested immunosuppressant potentials during in silico pipeline. In addition to estimation of free energies associated with ligand binding, MD simulation yielded us a great deal of information about ligand-macromolecule interactions to evaluate the pattern of interactions and the molecular basis of inhibition. The present study is a continuum of our efforts to identify interleukin-2 inhibitors of both natural and synthetic origin. Herein, we report molecular dynamics simulation studies of Interluekin-2 complexed with different antagonists previously reported by our group. The study of protein-ligand dynamics enabled us to gain a better understanding of the contribution of different active site residues in ligand binding. The results of the study will be used as the guide to rationalize the fragment based synthesis of drug-like interleukin-2 inhibitors as immune-modulators.

Keywords: immuno-modulators, MD simulation, protein-ligand interaction, structure-based drug design

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71 Vanadium (V) Complexes of a Tripodal Ligand, Their Characterization and Biological Implications

Authors: Mannar R. Maurya, Bhawna Uprety, Fernando Avecilla, Pedro Adão, J. Costa Pessoa

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The reaction of the tripodal tetradentate dibasic ligand 6,6'–(2–(pyridin–2–yl)ethylazanediyl)bis(methylene)bis(2,4–di–tert–butylphenol), H2L1 I, with [VIVO(acac)2] in CH3CN gives the VVO–complex, [VVO(acac)(L1)] 1. Crystallization of 1 in CH3CN at ~0 ºC, gives dark blue crystals of 1, while at room temperature it affords dark green crystals of [{VVO(L1)}2µ–O] 3. Upon prolonged treatment of 1 in MeOH [VVO(OMe)(MeOH)(L1)] 2 is obtained. All three complexes are analyzed by single–crystal X–ray diffraction, depicting distorted octahedral geometry around vanadium. In the reaction of H2L1 with VIVOSO4 partial hydrolysis of the tripodal ligand results in elimination of the pyridyl fragment of L1 and the formation of H[VVO2(L2)] 4, containing the ONO tridentate ligand 6,6'–azanediylbis(methylene)bis(2,4–di–tert–butylphenol), H2L2 II. Compound 4, which was not fully characterized, undergoes dimerization in acetone yielding the hydroxido–bridged [{VVO(L2)}2µ–(HO)2] 5, having distorted octahedral geometry around each vanadium. In contrast, from a solution of 4 in acetonitrile, the dinuclear compound [{VVO(L2)}2µ–O] 6 is obtained, with trigonal bipyramidal geometry around each vanadium. The methoxido complex 2 is successfully employed as a functional catechol–oxidase mimic in the oxidation of catechol to o–quinone under air. The process is confirmed to follow a Michaelis–Menten type kinetics with respect to catechol, the Vmax and KM values obtained being 7.66×10–6 M min -1 and 0.0557 M, respectively, and the turnover frequency is 0.0541 min–1. Complex 2 is also used as a catalyst precursor for the oxidative bromination of thymol in aqueous medium. The selectivity shows quite interesting trends, namely when not using excess of primary oxidizing agent, H2O2 the para mono–brominated product corresponds to ~93 % of the products and no dibromo derivative is formed.

Keywords: oxidovanadium (V) complexes, tripodal ligand, crystal structure, catechol oxidase mimetic activity

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70 Secondary Charged Fragments Tracking for On-Line Beam Range Monitoring in Particle Therapy

Authors: G. Traini, G. Battistoni, F. Collamati, E. De Lucia, R. Faccini, C. Mancini-Terracciano, M. Marafini, I. Mattei, S. Muraro, A. Sarti, A. Sciubba, E. Solfaroli Camillocci, M. Toppi, S. M. Valle, C. Voena, V. Patera

Abstract:

In Particle Therapy (PT) treatments a large amount of secondary particles, whose emission point is correlated to the dose released in the crossed tissues, is produced. The measurement of the secondary charged fragments component could represent a valid technique to monitor the beam range during the PT treatments, that is a still missing item in the clinical practice. A sub-millimetrical precision on the beam range measurement is required to significantly optimise the technique and to improve the treatment quality. In this contribution, a detector, named Dose Profiler (DP), is presented. It is specifically planned to monitor on-line the beam range exploiting the secondary charged particles produced in PT Carbon ions treatment. In particular, the DP is designed to track the secondary fragments emitted at large angles with respect to the beam direction (mainly protons), with the aim to reconstruct the spatial coordinates of the fragment emission point extrapolating the measured track toward the beam axis. The DP is currently under development within of the INSIDE collaboration (Innovative Solutions for In-beam Dosimetry in hadrontherapy). The tracker is made by six layers (20 × 20 cm²) of BCF-12 square scintillating fibres (500 μm) coupled to Silicon Photo-Multipliers, followed by two plastic scintillator layers of 6 mm thickness. A system of front-end boards based on FPGAs arranged around the detector provides the data acquisition. The detector characterization with cosmic rays is currently undergoing, and a data taking campaign with protons will take place in May 2017. The DP design and the performances measured with using MIPs and protons beam will be reviewed.

Keywords: fragmentation, monitoring, particle therapy, tracking

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69 Genetic Polymorphism of Milk Protein Gene and Association with Milk Production Traits in Local Latvian Brown Breed Cows

Authors: Daina Jonkus, Solvita Petrovska, Dace Smiltina, Lasma Cielava

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The beta-lactoglobulin and kappa-casein are milk proteins which are important for milk composition. Cows with beta-lactoglobulin and kappa-casein gene BB genotypes have highest milk crude protein and fat content. The aim of the study was to determinate the frequencies of milk protein gene polymorphisms in local Latvian Brown (LB) cows breed and analyze the influence of beta-lactoglobulin and kappa-casein genotypes to milk productivity traits. 102 cows’ genotypes of milk protein genes were detected using Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) and electrophoresis on 3% agarose gel. For beta-lactoglobulin were observed 2 types of alleles A and B and for kappa-casein 3 types: A, B and E. Highest frequency in beta-lactoglobulin gene was observed for B allele – 0.926. Molecular analysis of beta-lactoglobulin gene shows 86.3% of individuals are homozygous by B allele and animals are with genotypes BB and 12.7% of individuals are heterozygous with genotypes AB. The highest milk yield 4711.7 kg was for 1st lactation cows with AB genotypes, whereas the highest milk protein content (3.35%) and fat content (4.46 %) was for BB genotypes. Analysis of the kappa-casein locus showed a prevalence of the A allele – 0.750. The genetic variant of B was characterized by a low frequency – 0.240. Moreover, the frequency of E occurred in the LB cows’ population with very low frequency – 0.010. 54.9 % of cows are homozygous with genotypes AA, and only 4.9 % are homozygous with genotypes BB. 32.8 % of individuals are heterozygous with genotypes AB, and 2.0 % are with AE. The highest milk productivity was for 1st lactation cows with AB genotypes: milk yield 4620.3 kg, milk protein content 3.39% and fat content 4.53 %. According to the results, in local Latvian brown there are only 2.9% of cows are with BB-BB genotypes, which is related to milk coagulation ability and affected cheese production yield. Acknowledgment: the investigation is supported by VPP 2014-2017 AgroBioRes Project No. 3 LIVESTOCK.

Keywords: beta-lactoglobulin, cows, genotype frequencies, kappa-casein

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68 Unsupervised Classification of DNA Barcodes Species Using Multi-Library Wavelet Networks

Authors: Abdesselem Dakhli, Wajdi Bellil, Chokri Ben Amar

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DNA Barcode, a short mitochondrial DNA fragment, made up of three subunits; a phosphate group, sugar and nucleic bases (A, T, C, and G). They provide good sources of information needed to classify living species. Such intuition has been confirmed by many experimental results. Species classification with DNA Barcode sequences has been studied by several researchers. The classification problem assigns unknown species to known ones by analyzing their Barcode. This task has to be supported with reliable methods and algorithms. To analyze species regions or entire genomes, it becomes necessary to use similarity sequence methods. A large set of sequences can be simultaneously compared using Multiple Sequence Alignment which is known to be NP-complete. To make this type of analysis feasible, heuristics, like progressive alignment, have been developed. Another tool for similarity search against a database of sequences is BLAST, which outputs shorter regions of high similarity between a query sequence and matched sequences in the database. However, all these methods are still computationally very expensive and require significant computational infrastructure. Our goal is to build predictive models that are highly accurate and interpretable. This method permits to avoid the complex problem of form and structure in different classes of organisms. On empirical data and their classification performances are compared with other methods. Our system consists of three phases. The first is called transformation, which is composed of three steps; Electron-Ion Interaction Pseudopotential (EIIP) for the codification of DNA Barcodes, Fourier Transform and Power Spectrum Signal Processing. The second is called approximation, which is empowered by the use of Multi Llibrary Wavelet Neural Networks (MLWNN).The third is called the classification of DNA Barcodes, which is realized by applying the algorithm of hierarchical classification.

Keywords: DNA barcode, electron-ion interaction pseudopotential, Multi Library Wavelet Neural Networks (MLWNN)

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67 The Association between IFNAR2 and Dpp9 Genes Single Nucleotide Polymorphisms Frequency with COVID-19 Severity in Iranian Patients

Authors: Sima Parvizi Omran, Rezvan Tavakoli, Mahnaz Safari, Mohammadreza Aghasadeghi, Abolfazl Fateh, Pooneh Rahimi

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Background: SARS-CoV-2, a single-stranded RNA betacoronavirus causes the global outbreak of coronavirus disease 2019 (COVID-19). Several clinical and scientific concerns are raised by this pandemic. Genetic factors can contribute to pathogenesis and disease susceptibility. There are single nucleotide polymorphisms (SNPs) in many of the genes in the immune system that affect the expression of specific genes or functions of some proteins related to immune responses against viral infections. In this study, we analyzed the impact of polymorphism in the interferon alpha and beta receptor subunit 2 (IFNAR2) and dipeptidyl peptidase 9 (Dpp9) genes and clinical parameters on the susceptibility and resistance to Coronavirus disease (COVID-19). Methods: A total of 330- SARS-CoV-2 positive patients (188 survivors and 142 nonsurvivors) were included in this study. All single-nucleotide polymorphisms (SNPs) on IFNAR2 (rs2236757) and Dpp9 (rs2109069) were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: In survivor patients, the frequency of the favourable genotypes of IFNAR2 SNP (rs2236757 GC) was significantly higher than in nonsurvivor patients, and also Dpp9 (rs2109069 AT) genotypes were associated with the severity of COVID-19 infection. Conclusions: This study demonstrated that the severity of COVID- 19 patients was strongly associated with clinical parameters and unfavourable IFNAR2, Dpp9 SNP genotypes. In order to establish the relationship between host genetic factors and the severity of COVID-19 infection, further studies are needed in multiple parts of the world.

Keywords: SARS-CoV-2, COVID-19, interferon alpha and beta receptor subunit 2, dipeptidyl peptidase 9, single-nucleotide polymorphisms

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66 Comparative Assessment of Microplastic Pollution in Surface Water and Sediment of the Gomati and Saryu Rivers, India

Authors: Amit K. Mishra, Jaswant Singh

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The menace of plastic, which significantly pollutes the aquatic environment, has emerged as a global problem. There is an emerging concern about microplastics (MPs) accumulation in aquatic ecosystems. It is familiar to everyone that the ultimate end for most of the plastic debris is the ocean. Rivers are the efficient carriers for transferring MPs from terrestrial to aquatic, further from upstream to downstream areas, and ultimately to oceans. The root cause study can provide an effective solution to a problem; hence, tracing of MPs in the riverine system can illustrate the long-term microplastic pollution. This study aimed to investigate the occurrence and distribution of microplastic contamination in surface water and sediment of the two major river systems of Uttar Pradesh, India. One is the Gomti River, Lucknow, a tributary of the Ganga, and the second is the Saryu River, the lower part of the Ghagra River, which flows through the city of Ayodhya. In this study, the distribution and abundance of MPs in surface water and sediments of two rivers were compared. Samples of water and sediment were collected from different (four from each river) sampling stations in the river catchment of two rivers. Plastic particles were classified according to type, shape, and color. In this study, 1523 (average abundance 254) and 143 (average abundance 26) microplastics were identified in all studied sites in the Gomati River and Saryu River, respectively. Observations on samples of water showed that the average MPs concentration was 392 (±69.6) and 63 ((±18.9) particles per 50l of water, whereas the sediment sample showed that the average MPs concentration was 116 (±42.9) and 46 (±12.5) particles per 250gm of dry sediment in the Gomati River and Saryu River, respectively. The high concentration of microplastics in the Lucknow area can be attributed to human activities, population density, and the entry of various effluents into the river. Microplastics with fibrous shapes were dominated, followed by fragment shapes in all the samples. The present study is a pioneering effort to count MPs in the Gomati and Saryu River systems.

Keywords: freshwater, Gomati, microplastics, Saryu, sediment

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65 Fatty Acid Binding Protein 3 Gene Polymorphisms and Their Associations with Growth Traits and Blood Parameters in Two Iranian Sheep Breeds

Authors: Sahar Javadi-Novashnagh, Mohammad Moradi-Shahrbabak, Mostafa Sadeghi, Katarzyna Ropka-Molik, Hossein Moradi-Shahrbabak, Maria Consuelo Mura

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The objective of this study was to investigate two single nucleotide polymorphisms located in exon 2 (g.939A > G) and intron 3 (g.4349A > G) of fatty acid binding protein 3 (FABP3) gene in two Iranian sheep breeds, Lori-Bakhtiari and Zel, using polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) approach. The association of the polymorphisms with growth traits and blood parameters was also examined. Results revealed a g.939A > G SNP (single nucleotide polymorphism) in the exon 2 exhibiting three genotypes: AA, AG, and GG. Statistical analysis indicated that this polymorphism significantly influenced blood triglyceride (P < 0.05) and cholesterol (P < 0.08) levels as well as weaning weight (P < 0.05). Animals with AG genotype had the highest blood triglyceride level and weaning weight while the highest amount of blood cholesterol was observed in animals with GG genotype. On the other hand, no significant effect was observed on birth and fat-tail weight traits. The intron 3 (g.4349A > G) was monomorphic across the studied samples. Lori-Bakhtiari breed showed significantly higher blood triglyceride and cholesterol levels, as also birth and weaning weight compared to Zel breed (P < 0.01). Considering that the literature is bereft of any report on the association study between FABP3 SNPs and sheep growth traits and blood parameters, our findings suggest that the investigated polymorphism might be one of the main genetic factors affecting growth and physiological traits in sheep.

Keywords: FABP3 gene, fatness, weaning weight, blood triglyceride, cholesterol, Zel, Lori-Bakhtiari

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64 Studying the Photodegradation Behavior of Microplastics Released from Agricultural Plastic Products to the Farmland

Authors: Maryam Salehi, Gholamreza Bonyadinejad

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The application of agricultural plastic products like mulch, greenhouse covers, and silage films is increasing due to their economic benefits in providing an early and better-quality harvest. In 2015, the 4 million tons (valued a 10.6 million USD) global market for agricultural plastic films was estimated to grow by 5.6% per year through 2030. Despite the short-term benefits provided by plastic products, their long-term sustainability issues and negative impacts on soil health are not well understood. After their removal from the field, some plastic residuals remain in the soil. Plastic residuals in farmlands may fragment to small particles called microplastics (d<5mm). The microplastics' exposure to solar radiation could alter their surface chemistry and make them susceptible to fragmentation. Thus, this study examined the photodegradation of low density polyethylene as the model microplastics that are released to the agriculture farmland. The variation of plastic’s surface chemistry, morphology, and bulk characteristics were studied after accelerated UV-A radiation experiments and sampling from an agricultural field. The Attenuated Total Reflectance Fourier Transform Spectroscopy (ATR-FTIR) and X-ray Photoelectron Spectroscopy (XPS) demonstrated the formation of oxidized surface functional groups onto the microplastics surface due to the photodegradation. The Differential Scanning Calorimetry (DSC) analysis revealed an increased crystallinity for the photodegraded microplastics compared to the new samples. The gel permeation chromatography (GPC) demonstrated the reduced molecular weight for the polymer due to the photodegradation. This study provides an important opportunity to advance understanding of soil pollution. Understanding the plastic residuals’ variations as they are left in the soil is providing a critical piece of information to better estimate the microplastics' impacts on environmental biodiversity, ecosystem sustainability, and food safety.

Keywords: soil health, plastic pollution, sustainability, photodegradation

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63 DNA Polymorphism Studies of β-Lactoglobulin Gene in Native Saudi Goat Breeds

Authors: Amr A. El Hanafy, Muhammad I. Qureshi, Jamal Sabir, Mohamed Mutawakil, Mohamed M. Ahmed, Hassan El Ashmaoui, Hassan Ramadan, Mohamed Abou-Alsoud, Mahmoud Abdel Sadek

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β-Lactoglobulin (β-LG) is the dominant non-casein whey protein found in bovine milk and of most ruminants. The amino acid sequence of β-LG along with its 3-dimensional structure illustrates linkage with the lipocalin superfamily. Preliminary studies in goats indicated that milk yield can be influenced by polymorphism in genes coding for whey proteins. The aim of this study is to identify and evaluate the incidence of functional polymorphisms in the exonic and intronic portions of β-LG gene in native Saudi goat breeds (Ardi, Habsi, and Harri). Blood samples were collected from 300 animals (100 for each breed) and genomic DNA was extracted using QIAamp DNA extraction Kit. A fragment of the β-LG gene from exon 7 to 3’ flanking region was amplified with pairs of specific primers. Subsequent digestion with Sac II restriction endonuclease revealed two alleles (A and B) and three different banding patterns or genotypes i.e. AA, AB and BB. The statistical analysis showed that β-LG AA genotype had higher milk yield than β-LG AB and β-LG BB genotypes. Nucleotide sequencing of the selected β-LG fragments was done and submitted to GenBank NCBI (Accession No. KJ544248, KJ588275, KJ588276, KJ783455, KJ783456 and KJ874959). Two already established SNPs in exon 7 (+4601 and +4603) and one fresh SNP in the 3’ UTR region were detected in the β-LG fragments with designated AA genotype. The polymorphisms in exon 7 did not produce any amino acid change. Phylogenetic analysis on the basis of nucleotide sequences of native Saudi goats indicated evolutional similarity with the GenBank reference sequences of goat, Bubalus bubalis and Bos taurus.

Keywords: β-Lactoglobulin, Saudi goats, PCR-RFLP, functional polymorphism, nucleotide sequencing, phylogenetic analysis

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62 A Study of Growth Performance, Carcass Characteristic, Meat Quality and Association of Polymorphism in the ApoVLDL-II Gene with Fat Accumulation in the Female Broiler, Thai Native and Betong Chickens (KU Line)

Authors: C. Kridtayopas, W. Danvilai, P. Sopannarath, A. Kayan, W. Loongyai

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Both Betong chicken (KU Line) and Thai Native chickens were the high quality of the meat and low carcass fat compared to broiler chickens. The objective of this study was to determine the growth performance, carcass characteristic, meat quality and association of polymorphism in the ApoVLDL-II gene with fat accumulation in the female broiler, Thai Native and Betong (KU line) chickens at 4-14 weeks. The chickens were used and reared under the same environment and management (100 chicks per breed). The results showed that body weight (BW) of broiler chickens was significantly higher than Thai Native and Betong (KU line) chickens (P < 0.01) through all the experiment. At 4-8 weeks of age, feed conversion ratio (FCR) of broiler chickens was significantly better than Thai Native and Betong (KU line) chickens (P < 0.01), then increased at week 8-14. The percentage of breast, abdominal fat and subcutaneous fat of broiler chickens was significantly greater than Thai Native and Betong (KU line) chickens (P < 0.01). However, Thai Native chickens showed the highest percentage of liver (P < 0.01) when compared to other breeds. In addition, the percentage of wing of Thai Native and Betong (KU line) chickens were significantly (P < 0.01) higher than broiler chickens. Meat quality was also determined and found that, pH of breast meat left from slaughter 45 minutes (pH45) and 24 hours (pH24) of broiler was significantly higher than Thai Native and Betong (KU line) (P < 0.01) whereas the percentage of drip loss, thawing loss, cooking loss and shear force was not significantly different between breeds. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to genotype the polymorphism in the ApoVLDL-II gene in the broiler, Thai Native and Betong (KU line) chickens. The results found that, the polymorphism in the ApoVLDL-II gene at VLDL6 loci was not associated with fat accumulation in those studied population.

Keywords: ApoVLDL-II gene, Betong (KU line) chickens, broiler chickens, carcass characteristic, growth performance, meat quality, Thai native chickens

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61 The Subcellular Localisation of EhRRP6 and Its Involvement in Pre-Ribosomal RNA Processing in Growth-Stressed Entamoeba histolytica

Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E.histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

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60 Characterization of Banana Based Farming Systems in the Arumeru District, Arusha- Tanzania

Authors: Siah Koka, Rony Swennen

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Arumeru district is located in Arusha region in Upper Pangani basin in Tanzania. Economically it is dominated with agricultural activities. Banana, coffee, maize, beans, tomatoes, and cassava are the most important food and cash crops. This paper characterized the banana-based farming system of Arumeru district, evaluates its sustainability as well as research needs. The household questionnaire was performed on-site and on farm observation. Transect walk also involved to identify different agro- ecological zones. Results show that farm holdings (home gardens) are smaller than a hectare (0.7 ha) and continue to fragment as population continues to grow. Banana cultivation is the backbone of the farming systems present both in the upland and plains. In the upper belt banana found their place in the forest, which form the home garden structure typical to East African highland banana production systems. However, in the plains, cultivation is done in monoculture and depends heavily on irrigation. We found slightly less cultivars present and hypothetically more pest and disease pressure. This was mainly seen for Fusarium oxysporum species, which eradicates susceptible cultivars such as Mchare cultivars rapidly given the method of irrigation. The smaller permanent upland home garden plots provide thus a more suitable environment where banana perform better. It should be noted that findings indicated good performance to occur in the less suitable plains too. Good management is believed to be the most influencing factor, although our survey failed in identifying them. Population pressure is currently pushing the sustainable system in the uplands to its boundaries. Nutrient mining, deforestation and changing rain patterns threat production not only on Mt. Meru but on a global scale.

Keywords: Arumeru district, banana-based farming system, Tanzania, Arumeru district

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59 Designing Creative Events with Deconstructivism Approach

Authors: Maryam Memarian, Mahmood Naghizadeh

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Deconstruction is an approach that is entirely incompatible with the traditional prevalent architecture. Considering the fact that this approach attempts to put architecture in sharp contrast with its opposite events and transpires with attending to the neglected and missing aspects of architecture and deconstructing its stable structures. It also recklessly proceeds beyond the existing frameworks and intends to create a different and more efficient prospect for space. The aim of deconstruction architecture is to satisfy both the prospective and retrospective visions as well as takes into account all tastes of the present in order to transcend time. Likewise, it ventures to fragment the facts and symbols of the past and extract new concepts from within their heart, which coincide with today’s circumstances. Since this approach is an attempt to surpass the limits of the prevalent architecture, it can be employed to design places in which creative events occur and imagination and ambition flourish. Thought-provoking artistic events can grow and mature in such places and be represented in the best way possible to all people. The concept of event proposed in the plan grows out of the interaction between space and creation. In addition to triggering surprise and high impressions, it is also considered as a bold journey into the suspended realms of the traditional conflicts in architecture such as architecture-landscape, interior-exterior, center-margin, product-process, and stability-instability. In this project, at first, through interpretive-historical research method and examining the inputs and data collection, recognition and organizing takes place. After evaluating the obtained data using deductive reasoning, the data is eventually interpreted. Given the fact that the research topic is in its infancy and there is not a similar case in Iran with limited number of corresponding instances across the world, the selected topic helps to shed lights on the unrevealed and neglected parts in architecture. Similarly, criticizing, investigating and comparing specific and highly prized cases in other countries with the project under study can serve as an introduction into this architecture style.

Keywords: anti-architecture, creativity, deconstruction, event

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58 Development of a Microfluidic Device for Low-Volume Sample Lysis

Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman

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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.

Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet

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57 Worldbuilding as Critical Architectural Pedagogy

Authors: Jesse Rafeiro

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This paper discusses worldbuilding as a pedagogical approach to the first-year architectural design studio. The studio ran for three consecutive terms between 2016-2018. Taking its departure from the fifty-five city narratives in Italo Calvino’s Invisible Cities, students collectively designed in a “nowhere” space where intersecting and diverging narratives could be played out. Along with Calvino, students navigated between three main exercises and their imposed limits to develop architectural insight at three scales simulating the considerations of architectural practice: detail, building, and city. The first exercise asked each student to design and model a ruin based on randomly assigned incongruent fragments. Each student was given one plan fragment and two section fragments from different Renaissance Treatises. The students were asked to translate these in alternating axonometric projection and model-making explorations. Although the fragments themselves were imposed, students were free to interpret how the drawings fit together by imagining new details and atypical placements. An undulating terrain model was introduced in the second exercise to ground the worldbuilding exercises. Here, students were required to negotiate with one another to design a city of ruins. Free to place their models anywhere on the site, the students were restricted by the negotiation of territories marked by other students and the requirement to provide thresholds, open spaces, and corridors. The third exercise introduced new life into the ruined city through a series of design interventions. Each student was assigned an atypical building program suggesting a place for an activity, human or nonhuman. The atypical nature of the programs challenged the triviality of functional planning through explorations in spatial narratives free from preconceived assumptions. By contesting, playing out, or dreaming responses to realities taught in other coursework, this third exercise actualized learnings that are too often self-contained in the silos of differing course agendas. As such, the studio fostered an initial worldbuilding space within which to sharpen sensibility and criticality for subsequent years of education.

Keywords: architectural pedagogy, critical pedagogy, Italo Calvino, worldbuilding

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56 Phenotypic Characterization of Desi Naked Neck Chicken and Its Association with Insulin-Like Growth Factor-I (IGF-I) Gene Polymorphism in Pakistan

Authors: Akbar Nawaz Khan, Abdul Ghaffar, Muhammad Naeem Riaz

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The study was conducted to investigate the phenotypic features, morphometry and production potentialities of indigenous naked neck chicken (NN) of Pakistan under intensive management condition. A total of 35 NN chicks were randomly selected, and the experiment was performed at Poultry and wildlife research section NARC Islamabad for a period of 22 weeks. The predominant plumage color was black and golden while skin color was observed white. The average shank length, leg length, thigh length, keel length, chest breadth, head width, wing space, wing length, body length, body girth, body height and pubic bone width in adult males and females were 69.19 ± 3.34mm, 117.93 ± 4.42mm, 117.93 ± 4.42mm, 90.87 ± 6.53mm, 95.03 ± 4.56mm, 49.77 ± 2.53mm, 30.63 ± 1.50cm, 27.24 ± 2.71cm, 18.88 ± 0.65cm, 17.77 ± 1.01cm, 25.96 ± 0.56cm, 47.81 ± 1.41cm and 35.69 ± 4.09mm respectively. The average age and live body weight of NN chicken at sexual maturity were recorded as 165.85 days and 1269.38 g. While hen-day egg production of NN was recorded as 45%. The present study was aimed to investigate the existence of polymorphism at IGF-I gene in indigenous naked neck chicken through PCR based Restriction Fragment Length Polymorphism. Based on restriction analysis using Hinf I restriction enzyme, three genotypes were detected designated as AA, AC, and CC. Restriction analysis of PCR amplified product showed the presence of DNA fragments of 622, 378, 244 and 191, (genotypes). The PCR-RFLP analysis is easy, cost effective method which permits the easy characterization of IGF-I gene. This showed the investigated IGF-I genes can serve as good molecular markers for marker assisted selection (MAS) concerning growth related traits in chicken.

Keywords: Desi chicken, naked neck, morphology, morphometry, production potential, egg traits, egg geometry, IGF-I, growth, PCR- RFLP, chicken

Procedia PDF Downloads 357