Search results for: NOD2 (nucleotide binding oligomerization receptor 2)

Authors: Jaco Oosthuizen, Nerina C. Van Der Merwe

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The National Health Laboratory Services in Bloemfontien has been the diagnostic testing facility for 1830 patients for familial breast cancer since 1997. From the cohort, 540 were comprehensively screened using High-Resolution Melting Analysis or Next Generation Sequencing for the presence of point mutations and/or indels. Approximately 90% of these patients stil remain undiagnosed as they are BRCA1/2 negative. Multiplex ligation-dependent probe amplification was initially added to screen for copy number variation detection, but with the introduction of next generation sequencing in 2017, was substituted and is currently used as a confirmation assay. The aim was to investigate the viability of utilizing internationally designed copy number variation detection assays based on mostly European/Caucasian genomic data for use within a South African context. The multiplex ligation-dependent probe amplification technique is based on the hybridization and subsequent ligation of multiple probes to a targeted exon. The ligated probes are amplified using conventional polymerase chain reaction, followed by fragment analysis by means of capillary electrophoresis. The experimental design of the assay was performed according to the guidelines of MRC-Holland. For BRCA1 (P002-D1) and BRCA2 (P045-B3), both multiplex assays were validated, and results were confirmed using a secondary probe set for each gene. The next generation sequencing technique is based on target amplification via multiplex polymerase chain reaction, where after the amplicons are sequenced parallel on a semiconductor chip. Amplified read counts are visualized as relative copy numbers to determine the median of the absolute values of all pairwise differences. Various experimental parameters such as DNA quality, quantity, and signal intensity or read depth were verified using positive and negative patients previously tested internationally. DNA quality and quantity proved to be the critical factors during the verification of both assays. The quantity influenced the relative copy number frequency directly whereas the quality of the DNA and its salt concentration influenced denaturation consistency in both assays. Multiplex ligation-dependent probe amplification produced false positives due to ligation failure when ligation was inhibited due to a variant present within the ligation site. Next generation sequencing produced false positives due to read dropout when primer sequences did not meet optimal multiplex binding kinetics due to population variants in the primer binding site. The analytical sensitivity and specificity for the South African population have been proven. Verification resulted in repeatable reactions with regards to the detection of relative copy number differences. Both multiplex ligation-dependent probe amplification and next generation sequencing multiplex panels need to be optimized to accommodate South African polymorphisms present within the genetically diverse ethnic groups to reduce the false copy number variation positive rate and increase performance efficiency.

Keywords: familial breast cancer, multiplex ligation-dependent probe amplification, next generation sequencing, South Africa

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Authors: Gulcan Sahal, Isil Seyis Bilkay

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Candida species which often exist as commensal microorganisms in healthy individuals are major causes of important infections, especially in AIDS and immunocompromised patients, by means of their biofilm formation abilities. Therefore, in this study, determination of biofilm formation in different clinical strains of Candida species, investigation of strong biofilm forming Candida strains, examination of clinical information of each strong and weak biofilm forming Candida strains and investigation of some plant substances’ effects on biofilm formation of strong biofilm forming strains were aimed. In this respect, biofilm formation of Candida strains was analyzed via crystal violet binding assay. According to our results, biofilm levels of strains belong to different Candida species were different from each other. Additionally, it is also found that some plant substances effect biofilm formation. All these results indicate that, as well as C. albicans strains, other non-albicans Candida species also emerge as causative agents of infections and have biofilm formation abilities. In addition, usage of some plant substances in different concentrations may provide a new treatment against biofilm related Candida infections.

Keywords: anti-biofilm, biofilm formation, Candida species, biosystems engineering

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Authors: E. L. Albuquerque, U. L. Fulco, G. S. Ourique

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The focus of this work is on the numerical investigation of the charge transport properties of the de novo-designed alpha3 polypeptide, as well as in its variants, all of them probed by gene engineering. The theoretical framework makes use of a tight-binding model Hamiltonian, together with ab-initio calculations within quantum chemistry simulation. The alpha3 polypeptide is a 21-residue with three repeats of the seven-residue amino acid sequence Leu-Glu-Thr-Leu-Ala-Lys-Ala, forming an alpha–helical bundle structure. Its variants are obtained by Ala→Gln substitution at the e (5th) and g (7th) position, respectively, of the alpha3 polypeptide amino acid sequence. Using transmission electron microscopy and atomic force microscopy, it was observed that the alpha3 polypeptide and one of its variant do have the ability to form fibrous assemblies, while the other does not. Our main aim is to investigate whether or not the biased alpha3 polypeptide and its variants can be also identified by quantum charge transport measurements through current-voltage (IxV) curves as a pattern to characterize their fibrous assemblies. It was observed that each peptide has a characteristic current pattern, which may be distinguished by charge transport measurements, suggesting that it might be a useful tool for the development of biosensors.

Keywords: charge transport properties, electronic transmittance, current-voltage characteristics, biological sensor

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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Authors: Behrang Ekrami, Amin Tamadon, Iman Razeghian Jahromi, Darioush Moghadas, Mehdi Ghahremani-Seno, Mostafa Ghaderi-Zefrehei, Ahmad Sodagar Amiri, Taheri Reza

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Persian fallow deer (Dama dama mesopotamica) is belonging to the family Cervidae and is only found in a few protected areas in the northwest, north, and southwest of Iran. The aims of this study were analysis of inbreeding and morphometric parameters of semen in male Persian fallow deer to investigate the cause of reduced fertility of this endangered species in Dasht-e-Naz National Refuge, Sari, Iran. The Persian fallow deer semen was collected from four adult bucks randomly during the breeding and non-breeding season from five dehorned and horned deer's by an artificial vagina. Twelve blood samples was taken from Persian fallow deer and mitochondrial DNA was extracted, amplified, extracted, sequenced and then were considered for genetic analysis. The Persian fallow deer semen, both with normal and abnormal spermatozoa, is similar to that of domestic ruminants but very smaller and difficult to observe at the primary observation. The post-mating season collected ejaculates contained abnormal spermatozoa, debris and secretion of accessory glands in horned bucks and accessory glands secretion free of any spermatozoa in dehorned or early velvet budding bucks. Microscopic evaluation in all four bucks during the mating season showed the mean concentration of 9×106 spermatozoa/ml. The mean ± SD of age, testes length and testes width was 4.60 ± 1.52 years, 3.58 ± 0.32 and 1.86 ± 0.09 cm, respectively. The results identified 1120 loci (assuming each nucleotide as locus) in which 377 were polymorphic. In conclusion, reduced fertility of male Persian fallow deer may be caused by inbreeding of the protected herd in a limited area of Dasht-e-Naz National Refuge.

Keywords: Persian fallow deer, genetic analysis, spermatozoa, reproductive characteristics

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Authors: Jayanthi Sivaraman

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Claudins are the important components of the tight junctions that play a key role in paracellular permeability. Among various members of Claudin family, Claudin 4 (CLDN4) is found to be overexpressed in ovarian, pancreatic carcinomas and other epithelial malignancies. Therefore, in this study, an attempt has been made to identify potent inhibitors for CLDN4 from the ZINC database using virtual screening, molecular docking and molecular dynamics simulations. A well refined molecular model of CLDN4 was built using Prime of Schrodinger v10.2(Template- PDB ID: 4P79). Approximately, 6 million compounds from ZINC database are subjected to high-throughput virtual screening (HTVS) against the active site of CLDN4. Molecular docking using GLIDE predicted ARG31, ASN142, ASP146 and ARG158 as critically important residues. Furthermore, three compounds from ZINC database (ZINC96331839, ZINC36533519 and ZINC75819394) showed highly promising ADME properties and binding affinity with stable conformation. The therapeutic efficiency of these lead compounds is evaluated and confirmed by in-vitro and in-vivo studies which leads to the development of novel anti-cancer drugs.

Keywords: ADME property, inhibitors, molecular docking, virtual screening

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Authors: Seung-Hwa Baek, In-Jung Nam, Sang-Han Lee

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The development of anti-melanogenic agents is important for the prevention of serious esthetic problem like a melasma, freckle, age spots, and chloasma. The aim of this study was to investigate the anti-melanogenic effect of sesamol, an active lignan isolated from sesame seed, by mushroom and cellular tyrosinase assay, melanin content and the analysis of melanogensis-related mRNA expressions in melana cells. Sesamol showed strong inhibitory activity against the mushroom tyrosinase in a dose-dependent manner. Intracellular tyrosinase inhibition activity was also confirmed by zymography. At a concentration of 50 μM, sesamol inhibited melanin production in melan-a cells with no cytoxicity while those of phenylthiourea (PTU) as a positive control were the same condition. Sesamol significantly inhibited the expression of melanogensis-related genes, such as tyrosinase, tyrosinase-related protein-1 (TRP-1), dopachrome tautomerase (Dct), microphthalmia-associated transcription factor (MITF) and melanocortin 1 receptor (MC1R). These findings indicate that sesamol could reduce melanin biosynthesis via the downregulation of tyrosinase activity and melanin production via subsequent gene expression of melanogenesis-related proteins. Together, these results suggest that the sesamol have strong potential in inhibiting melanin biosynthesis, in that the substance may be used as a new skin-whitening agent of cosmetic materials.

Keywords: sesamol, sesame seed, melanin biosynthesis, melanogenesis-related gene, skin-whitening agent

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Authors: Othman E. Othman, Hassan R. Darwish, Amira M. Nowier

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Lactoferrin (LF) is a multifunctional protein involved in economically production traits like milk protein composition and skeletal structure in small ruminants including sheep and goat. So, LF gene - with its genetic polymorphisms associated with production traits - is considered a candidate genetic marker used in marker-assisted selection in goats. This study aimed to identify the different alleles and genotypes of this gene in three Egyptian goat breeds using PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing. Genomic DNA was extracted from 120 animals belonging to Barki, Zaraibi, and Damascus goat breeds. Using specific primers, PCR amplified 247-bp fragments from exon 2 of LF goat gene. The PCR products were subjected to Single-Strand Conformation Polymorphism (SSCP) technique. The results showed the presence of two genotypes GG and AG in the tested animals. The frequencies of both genotypes varied among the three tested breeds with the highest frequencies of GG genotype in all tested goat breeds. The sequence analysis of PCR products representing these two detected genotypes declared the presence of an SNP (single nucleotide polymorphisms) substitution (G/A) among G and A alleles of this gene. The association between different LF genotypes and milk composition as well as body measurement was estimated. The comparison showed that the animals possess AG genotypes are superior over those with GG genotypes for different parameters of milk protein compositions and skeletal structures. This finding declared that allele A of LF gene is considered the promising marker for the productive traits in goat. In conclusion, the Egyptian goat breeds will be needed to enhance their milk protein composition and growth trait parameters through the increasing of allele A frequency in their herds depending on the superior production traits of this allele in goats.

Keywords: lLactoferrin gene, PCR-SSCP, SNPs, Egyptian goat

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Authors: Aurelio Palacardo, Giulio Mangano, Alberto De Marco

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Nowadays, the global economic framework is extremely competitive, and it consequently requires an efficient deployment of the resources provided by EU. In this context, Project management practices are intended to be one of the levers for increasing such an efficiency. The objective of this work is to explore the usage of Project Management methodologies and good practices in the European-wide research program “Horizon2020” and establish whether their maturity might impact the project's success. This allows to identify strengths in terms of application of PM methodologies and good practices and, in turn, to provide feedback and opportunities for improvements to be implemented in future programs. In order to achieve this objective, the present research makes use of a survey-based data retrieval and correlation analysis to investigate the level of perceived PM maturity in H2020 projects and the correlation of maturity with project success. The results show the Project Managers involved in H2020 to hold a high level of PM maturity, confirming PM standards, which are imposed by the EU commission as a binding process, are effectively enforced.

Keywords: project management, project management maturity, maturity models, project success

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Authors: Anita Kushwaha

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We propose a new evolutionary computational model called Artificial Reproduction System which is based on the complex process of meiotic reproduction occurring between male and female cells of the living organisms. Artificial Reproduction System is an attempt towards a new computational intelligence approach inspired by the theoretical reproduction mechanism, observed reproduction functions, principles and mechanisms. A reproductive organism is programmed by genes and can be viewed as an automaton, mapping and reducing so as to create copies of those genes in its off springs. In Artificial Reproduction System, the binding mechanism between male and female cells is studied, parameters are chosen and a network is constructed also a feedback system for self regularization is established. The model then applies Mendel’s law of inheritance, allele-allele associations and can be used to perform data analysis of imbalanced data, multivariate, multiclass and big data. In the experimental study Artificial Reproduction System is compared with other state of the art classifiers like SVM, Radial Basis Function, neural networks, K-Nearest Neighbor for some benchmark datasets and comparison results indicates a good performance.

Keywords: bio-inspired computation, nature- inspired computation, natural computing, data mining

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Authors: Manar Makhoul, Buthainah Alsalamah, Salam Lawand, Hassan Azzam

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The major storage proteins in endosperm of 33 cultivated and wild barley genotypes (H.vulgare, H. spontaneum, H. bulbosum, H. murinum, H. marinum) were analyzed to demonstrate the variation in the hordein polypeptides encoded by multigene families in grains. The SDS-PAGE revealed 13 and 17 alleles at the Hor1 and the Hor2 loci respectively, with frequencies from 0.83 to 14 and 0.56 to 13.41% respectively, while seven alleles at the Hor3 locus with frequencies from 3.63 to 30.91% were recognized. The phylogenetic analysis indicated to relevance of the polymorphism in hordein patterns as successful tool in identifying the individual genotypes and discriminating the species according to genome type. We also reported in this research complete nucleotide sequence B-hordein genes of seven wild and cultivated barley genotypes. A 152bp upstream sequence of B-hordein promoter contained a TATA box, CATC box, AAAG motif, N-motif and E-motif. In silico analysis of B-Hordein sequences demonstrated that the coding regions were not interrupted by any intron, and included the complete ORF which varied between 882 and 906 bp, and encoded mature proteins with 293-301 residues characterized by high contents of glutamine (29%), and proline (18%). Comparison of the predicted polypeptide sequences with the published ones suggested that all S-rich prolamins genes are descended from common ancestor. The sequence started at N-terminal with a signal peptide, and then followed directly by two domains; a repetitive one based on the repetition of the repeat unit PQQPFPQQ and C-terminal domain. Also, it was found that positions of the eight cysteine residues were highly conserved in all the B-hordein sequences, but Hordeum bulbosum had additional unpaired one. The phylogenetic tree of B-hordein polypeptide separated the genotypes in distinct seven subgroups. In general, the high homology between B-hordeins and LMW glutenin subunits suggests similar bread-making influences for these B-hordeins.

Keywords: hordeum, phylogenetic tree, sequencing, storage protein

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Authors: Mariane G. Tadros, Shahira M. Ezzat, Maha M. Salama, Mohamed A. Farag

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In this study, the in vitro anticholinesterase activity of the volatile oils of both O. basilicum and O. africanum was investigated and both samples showed significant activity. As a result, the major constituents of the two oils were isolated using several column chromatography. Linalool, 1,8-cineol and eugenol were isolated from the volatile oil of O. basilicum and camphor was isolated from the volatile oil of O. africanum. The anticholinesterase activity of the isolated compounds were also evaluated where 1,8-cineol showed the highest inhibitory activity followed by camphor. To confirm these activities, learning and memory enhancing effects were tested in mice. Memory impairment was induced by scopolamine, a cholinergic muscarinic receptor antagonist. Anti-amnesic effects of both volatile oils and their terpenoids were investigated by the passive avoidance task in mice. We also examined their effects on brain acetylcholinesterase activity. Results showed that scopolamine-induced cognitive dysfunction was significantly attenuated by administration of the volatile oils and their terpenoids, eugenol and camphor, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that O. basilicum and O. africanum volatile oils can be good candidates for further studies on Alzheimer’s disease via their acetylcholinesterase inhibitory actions.

Keywords: Ocimum baselicum, Ocimum africanum, GC/MS analysis, anticholinesterase

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Authors: H. Oudira, A. Saifi

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The therapeutic utility of certain Auger emitters such as iodine-125 depends on their position within the cell nucleus . Or diagnostically, and to maintain as low as possible cell damage, it is preferable to have radionuclide localized outside the cell or at least the core. One solution to this problem is to consider markers capable of conveying anticancer drugs to the tumor site regardless of their location within the human body. The objective of this study is to simulate the impact of a complex such as bleomycin on single and double strand breaks in the DNA molecule. Indeed, this simulation consists of the following transactions: - Construction of BLM -Fe- DNA complex. - Simulation of the electron’s transport from the metastable state excitation of Fe 57 by the Monte Carlo method. - Treatment of chemical reactions in the considered environment by the diffusion equation. For physical, physico-chemical and finally chemical steps, the geometry of the complex is considered as a sphere of 50 nm centered on the binding site , and the mathematical method used is called step by step based on Monte Carlo codes.

Keywords: concentration, yield, radical species, bleomycin, excitation, DNA

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Authors: Ahmad Ali Shahid, Mukhtar Ahmed

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The demand for long staple fiber having better strength and length is increasing with the introduction of modern spinning and weaving industry in Pakistan. Work on gene discovery from developing cotton fibers has helped to identify dozens of genes that take part in cotton fiber development and several genes have been characterized for their role in fiber development. Sucrose synthase (SuS) is a key enzyme in the metabolism of sucrose in a plant cell, in cotton fiber it catalyzes a reversible reaction, but preferentially converts sucrose and UDP into fructose and UDP-glucose. UDP-glucose (UDPG) is a nucleotide sugar act as a donor for glucose residue in many glycosylation reactions and is essential for the cytosolic formation of sucrose and involved in the synthesis of cell wall cellulose. The study was focused on successful Agrobacterium-mediated stable transformation of SuS gene in pCAMBIA 1301 into cotton under a CaMV35S promoter. Integration and expression of the gene were confirmed by PCR, GUS assay, and real-time PCR. Young leaves of SuS overexpressing lines showed increased total soluble sugars and plant biomass as compared to non-transgenic control plants. Cellulose contents from fiber were significantly increased. SEM analysis revealed that fibers from transgenic cotton were highly spiral and fiber twist number increased per unit length when compared with control. Morphological data from field plants showed that transgenic plants performed better in field conditions. Incorporation of genes related to cotton fiber length and quality can provide new avenues for fiber improvement. The utilization of this technology would provide an efficient import substitution and sustained production of long-staple fiber in Pakistan to fulfill the industrial requirements.

Keywords: agrobacterium-mediated transformation, cotton fiber, sucrose synthase gene, staple length

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Authors: Siddharth Vishwakarma, Danie Shajie A., Mishra H. N.

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Milk powder becomes very useful in the low milk supply area but the exact amount to add for one glass of milk and the handling is difficult. So, the idea of instant soluble milk tablet comes into existence for its high solubility and easy handling. The moisture content of milk tablets is increased by the direct addition of water with no additives for binding. The variation of the tensile strength of instant soluble milk tablets and the flowability of milk powder with the moisture content is analyzed and optimized for the highest tensile strength of instant soluble milk tablets and flowability, above a particular value of milk powder using response surface methodology. The flowability value is necessary for ease in quantifying the milk powder, as a feed, in the designed tablet making machine. The instant soluble nature of milk tablets purely depends upon the disintegration characteristic of tablets in water whose study is under progress. Conclusions: The optimization results are very useful in the commercialization of milk tablets.

Keywords: flowability, milk powder, response surface methodology, tablet making machine, tensile strength

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Authors: Shri Krishna Mishra, Badri Yadav

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Proficient persons should be involved in formulation of the structure of the textbooks so that the topics selected in the Hindi textbooks for Class VII should contribute towards linguistic and literary development of the child and the language of the textbook matches the comprehension level of the student.The topics of tile textbooks should provide good illustrations and suitable exercises. Topics of variety of taste can be included in the textbook to satisfy the inquisitive children. There could be abstracts/hints at the beginning of each lesson. Meanings for difficult words must be given at the end of each topic for convenience of the parents and children as most of them find it difficult and time consuming to use Hindi dictionary. Exercises should be relevant covering the whole topic and the difficulty level should match the maturity level of the students in respect of CBSE Board. The stitching and binding of CBSE prescribed books may be improved to increase durability.

Keywords: comparative study of CBSE and BSEMPB, Central Secondary Education, Board of Secondry Education, BHOPAL

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Authors: AbdelRahman A. Faragalla, Mohamed H. Alqhtani, Mohamed M. M.Ahmed

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Saudi Arabia has currently been beset by a barrage of bizarre assemblages of subterranean termite fauna, inflicting heavy catastrophic havocs on human valued properties in various homes, storage facilities, warehouses, agricultural and horticultural crops including okra, sweet pepper, tomatoes, sorghum, date palm trees, citruses and many forest domains and green lush desert oases. The most pressing urgent priority is to use modern technologies to alleviate the painstaking obstacle of taxonomic identification of these injurious noxious pests that might lead to effective pest control in both infested agricultural commodities and field crops. Our study has indicated the use of DNA fingerprinting technologies, in order to generate basic information of the genetic similarity between 3 predominant families containing the most destructive termite species. The methodologies included extraction and DNA isolation from members of the major families and the use of randomly selected primers and PCR amplifications with the nucleotide sequences. GC content and annealing temperatures for all primers, PCR amplifications and agarose gel electrophoresis were also conducted in addition to the scoring and analysis of Random Amplification Polymorphic DNA-PCR (RAPDs). A phylogenetic analysis for different species using statistical computer program on the basis of RAPD-DNA results, represented as a dendrogram based on the average of band sharing ratio between different species. Our study aims to shed more light on this intriguing subject, which may lead to an expedited display of the kinship and relatedness of species in an ambitious undertaking to arrive at correct taxonomic classification of termite species, discover sibling species, so that a logistic rational pest management strategy could be delineated.

Keywords: DNA fingerprinting, Western Saudi Arabia, DNA primers, RAPD

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Authors: Hui-Ling Huang, Tamara Vasylenko, Phasit Charoenkwan, Shih-Hsiang Chiu, Shinn-Ying Ho

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The knowledge of the human transporters is still limited due to technically demanding procedure of crystallization for the structural characterization of transporters by spectroscopic methods. It is desirable to develop bioinformatics tools for effective analysis of available sequences in order to identify human transmembrane transporter proteins (HMTPs). This study proposes a scoring card method (SCM) based method for predicting HMTPs. We estimated a set of propensity scores of dipeptides to be HMTPs using SCM from the training dataset (HTS732) consisting of 366 HMTPs and 366 non-HMTPs. SCM using the estimated propensity scores of 20 amino acids and 400 dipeptides -as HMTPs, has a training accuracy of 87.63% and a test accuracy of 66.46%. The five top-ranked dipeptides include LD, NV, LI, KY, and MN with scores 996, 992, 989, 987, and 985, respectively. Five amino acids with the highest propensity scores are Ile, Phe, Met, Gly, and Leu, that hydrophobic residues are mostly highly-scored. Furthermore, obtained propensity scores were used to analyze physicochemical properties of human transporters.

Keywords: dipeptide composition, physicochemical property, human transmembrane transporter proteins, human transmembrane transporters binding propensity, scoring card method

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Authors: M. Hosseinnezhad, K. Gharanjig

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Natural dyes, extracted from black carrot and bramble, were utilized as photosensitizers to prepare dye-sensitized solar cells (DSSCs). Spectrophotometric studies of the natural dyes in solution and on a titanium dioxide substrate were carried out in order to assess changes in the status of the dyes. The results show that the bathochromic shift is seen on the photo-electrode substrate. The chemical binding of the natural dyes at the surface photo-electrode were increased by the chelating effect of the Ti(IV) ions. The cyclic voltammetry results showed that all extracts are suitable to be performed in DSSCs. Finally, photochemical performance and stability of DSSCs based on natural dyes were studied. The DSSCs sensitized by black carrot extract have been reported to achieve up to Jsc=1.17 mAcm^-2, Voc= 0.55 V, FF= 0.52, η=0.34%, whereas Bramble extract can obtain up to Jsc=2.24 mAcm^-2, Voc= 0.54 V, FF= 0.57, η=0.71%. The power conversion efficiency was obtained from the mixed dyes in DSSCs. The power conversion efficiency of dye-sensitized solar cells using mixed Black carrot and Bramble dye is the average of the their efficiency in single DSSCs.

Keywords: anthocyanin, dye-sensitized solar cells, green energy, optical materials

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Authors: Mastaneh Safarnejad Shad, Wim Dehaen, Steven De Jonghe

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Since CXCR4 is the main coreceptor of HIV-1 and plays an important role in human immunodeficiency virus (HIV) entry, numerous efforts were directed towards the discovery of new classes of small molecules that act as CXCR4 antagonists. In addition, CXCR4 antagonists are potentially useful in the treatment of several other disorders, such as cancer cell metastasis, leukemia cell proliferation, rheumatoid arthritis, and pulmonary fibrosis. Since AMD3100 (plerixafor) is the only CXCR4 antagonist which obtained approval by the Food and Drug Administration (FDA), we were motivated to investigate a new category of molecules as CXCR4 antagonists. Most of the scaffolds which have been studied so far as CXCR4 antagonists are based on the tetrahydroquinoline (THQ) moiety in which AMD11070 (mavorixafor), GSK-812394, and TIQ15 displayed the most potent CXCR4 antagonism. Due to the high potency of these scaffolds, two different series of compounds were prepared in this work. In the first set, the THQ moiety is coupled to an amine chain and various isoquinoline derivatives (prepared by an in-house developed triazolization strategy), of which the upper part of molecules is identical to AMD11070 and TIQ15. In the second category of compounds, the THQ moiety was simplified by the synthesis of a substituted pyridine moiety. In order to investigate if CXCR4 antagonism requires the presence of an isoquinoline moiety, the corresponding pyridine analogues were also prepared. In both series of compounds, potent CXCR4 antagonism was noticed.

Keywords: CXCR4 coreceptor, CXCR4 antagonists, HIV inhibitor, tetrahydroquinoline

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Authors: Kyung Min Kwom, Young Joo Lee

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Purpose: Up to 90% of pancreatic cancer patient suffer from neuropathic pain. In palliative care setting, pain control in a pancreatic cancer patient is one of the major goals. Ketamine is a NMDA receptor antagonist effective in neuropathic pain. Also, there have been studies about opioid sparing effect of ketamine. This study was held in palliative care unit among pancreatic cancer patients to find out the factors related to ketamine use and the opioid sparing effect. Methods: Medical records of pancreatic cancer patients admitted to St. Mary’s hospital palliative care unit from 2013.1 to 2014.12 were reviewed. Patients were divided into two categories according to ketamine use. Also, opioid use before and after ketamine use was compared in ketamine group. Results: Compared to non ketamine use group, patients in ketamine group required a higher dose of opioid. Total opioid dose, daily opioid dose, number of daily rescue medication, daily average rescue dose were statistically significantly higher in ketamine group. Opioid requirement was increased after ketamine administration. Conclusion: In this study, ketamine group required more opioid. Ketamine is frequently considered in patients with severe pain, requiring high amount of opioid. Also, ketamine did not have an opioid sparing effect. Future studies about palliative use of ketamine in a larger number of patients are required.

Keywords: ketamine, opioid sparing, palliative care, pancreatic cancer

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Authors: Flutura Tahiraj, Emine Abdyli

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As a new alternative, mediation is integrated in the legislation of both developed and developing countries in Europe. Various researches in member states of the Council of Europe revealed obstacles, particularly related to the implementation of mediation in criminal matters. They are addressed through several recommendations and non-binding guidelines. However, there is limited empirical research on how the mediation in criminal matters is being implemented in the contexts of developing countries in South-Eastern Europe. Hence, the purpose of this qualitative study is to assess mediation in criminal matters in Kosovo by exploring how the main stakeholders describe the legal basis and implementation process and what it indicates for future practices. The data were gathered through 11 semi-structured interviews with judges, prosecutors, mediation clerks and mediators. Results show that laws and other guidelines that have been introduced since 2008 constitute a solid legal ground that facilitates mediation in criminal matters. The stakeholders are well aware of benefits mediation brings and express their willingness to advance its application to criminal matters. Results also indicate uncertainty among judges and prosecutors regarding the assessment and referral of certain criminal offences to mediation. To address it, specialized trainings, exchange programs and continuous monitoring and evaluation of the process could be supportive.

Keywords: mediation in criminal matters, legislation, implementation of mediation

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Authors: Jake Gonzalez, Tommy Dang

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This paper proposes a temporal graph approach to visualize and analyze the evolution of gene relationships and nomenclature over time. An interactive web-based tool implements this temporal graph, enabling researchers to traverse a timeline and observe coupled dynamics in network topology and naming conventions. Analysis of a real human genomic dataset reveals the emergence of densely interconnected functional modules over time, representing groups of genes involved in key biological processes. For example, the antimicrobial peptide DEFA1A3 shows increased connections to related alpha-defensins involved in infection response. Tracking degree and betweenness centrality shifts over timeline iterations also quantitatively highlight the reprioritization of certain genes’ topological importance as knowledge advances. Examination of the CNR1 gene encoding the cannabinoid receptor CB1 demonstrates changing synonymous relationships and consolidating naming patterns over time, reflecting its unique functional role discovery. The integrated framework interconnecting these topological and nomenclature dynamics provides richer contextual insights compared to isolated analysis methods. Overall, this temporal graph approach enables a more holistic study of knowledge evolution to elucidate complex biology.

Keywords: temporal graph, gene relationships, nomenclature evolution, interactive visualization, biological insights

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Authors: S. Ghalem, M. Mesmoudi, I. Daoudand, H. Allali

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The nitrogen-containing bisphosphonates (N-BPs) are well established as the treatments of choice for disorders of excessive bone resorption, myeloma and bone metastases, and osteoporosis. They inhibit farnesyl pyrophosphate synthase (FFPS), a key enzyme in the mevalonate pathway, resulting in inhibition of the prenylation of small GTP-binding proteins in osteoclasts and disruption of their cytoskeleton, adhesion/spreading, and invasion of cancer cells. A very few examples for synthesis of α-amino bisphosphonates based on several amino acids are known from the literature. In the present work, esters of aminoacid react with ketophsophonate (or their analog acid or acyl) to afford the desired products, α-iminophosphonates. The reaction of imine with dimethyl phosphate in the presence of catalytic amount of I2 give ester of α-aminobisphosphonate as sole product in good yield. Finally, we used computational docking methods to predict how several α-aminobisphosphonates bind to FPPS and how R and X influence. Pamidronate, β-aminobisphosphonate already marketed, was used as reference. These results are of interest since they represent a new and simple way to sythesize α-aminobisphosphonates with a free COOH group increased by R2 functionalisable and opening up the possibility of using the molecular docking to facilitate the design of other, novel FFPS inhibitors.

Keywords: drug research, cancer, α-amino bisphosphonates, molecular docking

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Authors: Zeyu Zhou, Ziyu Zhao, Xiaochen Yang, Ling AI, Heng Zhai, Stuart Holmes

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As a promising sustainable alternative to traditional fossil fuels, fuel cell technology is highly favoured due to its enhanced working efficiency and reduced emissions. In the context of high-temperature fuel cells (operating above 100 °C), the most commonly used proton exchange membrane (PEM) is the Polybenzimidazole (PBI) doped phosphoric acid (PA) membrane. Grafting is a promising strategy to advance PA-doped PBI PEM technology. The existing grafting modification on PBI PEMs mainly focuses on grafting phosphate-containing or alkaline groups onto the PBI molecular chains. However, quaternary ammonium-based grafting approaches face a common challenge. To initiate the N-alkylation reaction, deacidifying agents such as NaH, NaOH, KOH, K2CO3, etc., can lead to ionic crosslinking between the quaternary ammonium group and PBI. Polyvinylpyrrolidone (PVP) is another widely used polymer, the N-heterocycle groups within PVP endow it with a significant ability to absorb PA. Recently, PVP has attracted substantial attention in the field of fuel cells due to its reduced environmental impact and impressive fuel cell performance. However, due to the the poor compatibility of PVP in PBI, few research apply PVP in PA-doped PBI PEMs. This work introduces an innovative strategy to graft PVP onto PBI to form a network-like polymer. Due to the absence of quaternary ammonium groups, PVP does not pose issues related to crosslinking with PBI. Moreover, the nitrogen-containing functional groups on PVP provide PBI with a robust phosphoric acid retention ability. The nuclear magnetic resonance (NMR) hydrogen spectrum analysis results indicate the successful completion of the grafting reaction where N-alkylation reactions happen on both sides of the grafting agent 1,4-bis(chloromethyl)benzene. On one side, the reaction takes place with the hydrogen atoms on the imidazole groups of PBI, while on the other side, it reacts with the terminal amino group of PVP. The XPS results provide additional evidence from the perspective of the element. On synthesized PBI-g-PVP surfaces, there is an absence of chlorine (chlorine in grafting agent 1,4-bis(chloromethyl)benzene is substituted) element but a presence of sulfur element (sulfur element in terminal amino PVP appears in PBI), which demonstrates the occurrence of the grafting reaction and PVP is successfully grafted onto PBI. Prepare these modified membranes into MEA. It was found that during the fuel cell operation, all the grafted membranes showed substantial improvement in maximum current density and peak power density compared to unmodified one. For PBI-g-PVP 30, with a grafting degree of 22.4%, the peak power density reaches 1312 mW cm⁻², marking a 59.6% enhancement compared to the pristine PBI membrane. The improvement is caused by the improved PA binding ability of the membrane after grafting. The AST test result shows that the grafting membranes have better long-term durability and performance than unmodified membranes attributed to the presence of added PA binding sites, which can effectively prevent the PA leaching caused by proton migration. In conclusion, the test results indicate that grafting PVP onto PBI is a promising strategy which can effectively improve the fuel cell performance.

Keywords: fuel cell, grafting modification, PA doping ability, PVP

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Authors: A. Chegenizadeh, M. Keramatikerman, H. Nikraz

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Bitumen is one of the most applicable materials in pavement engineering. It is a binding material with unique viscoelastic properties, especially when it mixes with polymer. In this study, to figure out the viscoelastic behaviour of the polymer modified with bitumen (PMB), a series of dynamic shearing rheological (DSR) tests were conducted. Four percentages of lime (i.e. 1%, 2%, 4% and 5%) were mixed with PMB and tested under four different temperatures including 64ºC, 70ºC, 76ºC and 82ºC. The results indicated that complex shearing modulus (G*) increased by increasing the frequency due to raised resistance against deformation. The phase angle (δ) showed a decreasing trend by incrementing the frequency. The addition of lime percentages increased the complex modulus value and declined phase angle parameter. Increasing the temperature decreased the complex modulus and increased the phase angle until 70ºC. The decreasing trend of rutting factor with increasing temperature revealed that rutting factor improved by the addition of the lime to the PMB.

Keywords: rheological properties, DSR test, polymer mixed with bitumen (PMB), complex modulus, lime

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Authors: Nemat Sokhandan Bashir

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Threats from viruses to agricultural crops could be even larger than the losses caused by the other pathogens because, in many cases, the viral infection is latent but crucial from an epidemic point of view. Wild vegetation can be a source of many viruses that eventually find their destiny in crop plants. Although often asymptomatic in wild plants due to adaptation, they can potentially cause serious losses in crops. Therefore, exploring viruses in wild vegetation is very important. Recently, omics have been quite useful for exploring plant viruses from various plant sources, especially wild vegetation. For instance, we have discovered viruses such as Ambrossia asymptomatic virus I (AAV-1) through the application of metagenomics from Oklahoma Prairie Reserve. Accordingly, extracts from randomly-sampled plants are subjected to high speed and ultracentrifugation to separated virus-like particles (VLP), then nucleic acids in the form of DNA or RNA are extracted from such VLPs by treatment with phenol—chloroform and subsequent precipitation by ethanol. The nucleic acid preparations are separately treated with RNAse or DNAse in order to determine the genome component of VLPs. In the case of RNAs, the complementary cDNAs are synthesized before submitting to DNA sequencing. However, for VLPs with DNA contents, the procedure would be relatively straightforward without making cDNA. Because the length of the nucleic acid content of VPLs can be different, various strategies are employed to achieve sequencing. Techniques similar to so-called "chromosome walking" may be used to achieve sequences of long segments. When the nucleotide sequence data were obtained, they were subjected to BLAST analysis to determine the most related previously reported virus sequences. In one case, we determined that the novel virus was AAV-l because the sequence comparison and analysis revealed that the reads were the closest to the Indian citrus ringspot virus (ICRSV). AAV—l had an RNA genome with 7408 nucleotides in length and contained six open reading frames (ORFs). Based on phylogenies inferred from the replicase and coat protein ORFs of the virus, it was placed in the genus Mandarivirus.

Keywords: wild, plant, novel, metagenomics

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Authors: Mishari Khajah, Mohd. Elmuntasir Ahmed, Abdullah Al-Matouq, Farah Al-Ajeel, Fatemah Dashti, Ahmed Shishter

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The overall aim of this study was to monitor and evaluate the effluent quality of a reverse osmosis (RO) reject wastewater from the biggest wastewater treatment plant in the world that is using RO and ultrafiltration membranes in their processes to reclaim water for indirect potable water reuse from municipal wastewaters. The RO reject wastewater or brine included various contaminants that could harm the human health and the environment such as trace organics, organic matters, heavy metals, nutrients and pathogens. Unfortunately, there are no legally binding regulatory guidelines for brine management in Kuwait as many countries around the world. This study monitors and evaluate the RO reject wastewater (brine) generated from the Sulaibiya Wastewater Treatment Plant. Samples were collected and analyzed about 37 parameters for one-year period, twice a month, and compare it to Kuwait Environment Public Authority, KEPA. Results showed that the heavy metals parameters were above KEPA standards, which needs to be treated.

Keywords: domestic wastewater, management, potable water, RO reject wastewater, Sulaibiya wastewater treatment plant

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Authors: Olumuyiwa I. Ojo, Kola Amao, Josiah A. Adeyemo, Fred A. O. Otieno

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A study was carried out to assess the environmental impact of Kampe (Omi) irrigation scheme with respect to public health hazards, the rising water table, salinity and alkalinity problems on the project site. A structured questionnaire was used as the main tool to gather information on the effect of the irrigation project on the various communities around the project site. The different sections of the questionnaire enabled the gathering of information ranging from general to more specific information. The results obtained from the study showed that the two effects are obvious: the 'positive effects' which include increasing the socioeconomic development of the entire communities, resulting in an increase in employment opportunities and better lifestyle and the 'negative effects' in which malaria (100% occurrence) and schistosomiasis (66.7%) were found to be active diseases caused by irrigation activities. Increase in height of water table and salinity is eminent in the irrigation site unless adequate drainage is provided. The collection and experimental analyses of representation soil and water samples from each scheme were used to assess the current status of each receptor. Results obtained indicate the absence of soil with sodium adsorption ration (SAR) values ranging from 3.0 to 3.89, exchangeable sodium percentage (ESP) ranged from 3.8% to 5.5% while pH values ranged from 6.60 to 7.00. Drainage facilities of the project site are inadequate, therefore making it difficult to leach the soil and flood history is occasional.

Keywords: irrigation, impact, soil analysis, Nigeria

Procedia PDF Downloads 294

Authors: Nafiah Solikhah Thoha

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One area that has spatial character as Heritage area is Kauman Villages. Kauman village in The City of Surakarta, Central Java, Indonesia was formed in 1757 by Paku Buwono III as the King of Kasunanan kingdom (Mataram Kingdom) for Kasunanan kingdom courtiers and scholars of Madrasa. Spatial character of Kauman village influenced by Islamic planning and socio-cultural rules of Kasunanan Kingdom. As traditional settlements influenced by Islamic planning, the Grand Mosque is a binding part of the whole area. Circulation pattern forming network (labyrinth) with narrow streets that ended at the Grand Mosque. The outdoor space can be used for circulation. Social activity is dominated by step movement from one place to a different place. Stalemate (the fina/cul de sac) generally only passable on foot, bicycles, and motorcycles. While the pass (main and branch) can be traversed by motor, vehicles. Kauman village has an area that can not be used as a public road that penetrates and serves as a liaison between the outside world to the other. Hierarchy of hall in Kauman village shows that the existence of a space is getting into more important. Firstly, woman in Kauman make the handmade batik for themself. In 2005 many people improving batik tradisional into commercial, and developed program named "Batik Tourism village of Kauman". That program affects the spatial transformations. This study aimed to explore the influence of tourism program towards spatial transformations. The factors that studied are the organization of space, circulation patterns, hierarchical space, and orientation through the descriptive-evaluation approach methods. Based on the study, tourism activity engenders transformations on the spatial scale (macro), residential block (mezo), homes (micro). First, the Grand Mosque and madrasa (religious school) as a binding zoning; tangle of roads as forming the structure of the area developed as a liaison with outside Kauman; organization of space in the residential of batik entrepreneurs firstly just a residential, then develop into residential, factory of batik including showroom. Second, the circulation pattern forming network (labyrinth) and ends at the Grand Mosque. Third, the hierarchy in the form of public space (the shari), semi-public, and private (the fina/culdesac) is no longer to provide protection to women, only as hierarchy of circulation path. Fourth, cluster building orientation does not follow the kiblat direction or axis oriented to cosmos, but influence by the new function as the showroom. It was need the direction of the main road. Kauman grow as an appropriate area for the community. During its development, the settlement function changes according to community activities, especially economic activities. The new function areas as tourism area affect spatial pattern of Kauman village. Spatial existence and activity as a local wisdom that has been done for generations have meaning of holistic, encompassing socio-cultural sustainability, economics, and the heritage area. By reviewing the local wisdom and the way of life of that society, we can learn how to apply the culture as education for sustainable of heritage area.

Keywords: impact of tourism, Kauman village, spatial transformation, sustainable of heritage area

Procedia PDF Downloads 430