Search results for: site specificity

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: Samia Ait Ali Yahia

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Rock paintings and engravings in the Kabylia region of Algeria have been extensively studied, with 54 sites identified. These artworks were primarily discovered by Poyto and Musso in the mid-1960s. The paintings are predominantly adorned with red ochre ornaments, while some engravings can also be found on sandstone rocks. These artistic expressions can be found in various locations, such as shelters, rocks, and sandstone blocks in the northern part of Kabylia. These sites showcase a diverse range of decorations, including human figures, animal silhouettes, enigmatic designs, symbolic drawings, engravings, and Libyan characters. The research will involve conducting fieldwork at the Azru Imeyazen site to identify and study the different paintings and engravings present. This research aims to provide a detailed description of the rock paintings and engravings found in Kabylia, specifically focusing on the Azru Imeyazen (Tarihant) site.

Keywords: rock paintings, engraving, Kabylia, Tarihant, Azru Imayazen

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Authors: Amar Ranjan, Julieana Durai, Pranay Tanwar

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Background &Introduction: Ovarian cancer is one of the most common malignant tumor in the female. 70% of the cases of ovarian cancer are diagnosed at an advanced stage. The five-year survival rate associated with ovarian cancer is less than 30%. The early diagnosis of ovarian cancer becomes a key factor in improving the survival rate of patients. Presently, CAl25 (carbohydrate antigen125) is used for the diagnosis and therapeutic monitoring of ovarian cancer, but its sensitivity and specificity is not ideal. The introduction of HE4, human epididymis protein 4 has attracted much attention. HE4 has a sensitivity and specificity of 72.9% and 95% for differentiating between benign and malignant adnexal masses, which is better than CA125 detection.  Methods: Serum HE4 and CA -125 were estimated using the chemiluminescence method. Our cases were 40 epithelial ovarian cancer, 9 benign ovarian tumor, 29 benign gynaecological diseases and 13 healthy individuals. This group include healthy woman those who have undergoing family planning and menopause-related medical consultations and they are negative for ovarian mass. Optimal cut off values for HE4 and CA125 were 55.89pmol/L and 40.25U/L respectively (determined by statistical analysis). Results: The level of HE4 was raised in all ovarian cancer patients (n=40) whereas CA125 levels were normal in 6/40 ovarian cancer patients, which were the cases of OC confirmed by histopathology. There is a significant decrease in the level of HE4 with comparison to CA125 in benign ovarian tumor cases. Both the levels of HE4 and CA125 were raised in the nonovarian cancer group, which includes cancer of endometrium and cervix. In the healthy group, HE4 was normal in all patients except in one case of the rudimentary horn, and the reason for this raised HE4 level is due to the incomplete development of uterus whereas CA125 was raised in 3 cases. Conclusions: Findings showed that the serum level of HE4 is an important indicator in the diagnosis of ovarian cancer, and it also distinguishes between benign and malignant pelvic masses. However, a combination of HE4 and CA125 panel will be extremely valuable in improving the diagnostic efficiency of ovarian cancer. These findings of our study need to be validated in the larger cohort of patients.

Keywords: human epididymis protein 4, ovarian cancer, diagnosis, benign lesions

Procedia PDF Downloads 128

Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity

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Authors: Mohammed Aljoma, Eblal Zakzok

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Safety is a science that seeks to protect and avoid humans from risks in any field and prevent losses in properties and lives as much as possible. On the other hand, occupational safety goals aim to protect workers from risks which can occur during work execution. The main purpose of occupational safety is to ultimately protect people, properties and the environment by reducing accidents and injuries that may cause losses and damages. To achieve this goal, we must remove the direct and indirect reasons which cause accidents and injuries; some of the reasons of accidents are the unsafe cases and inept behavior or both of them. This research focuses on the manner of providing instant protection from the very first beginning to people, properties and the environment by: -Inserting safety demands in the planning and designing works by identifying risk levels in every task of the project, -Using a new risk managing system or modifying or changing a previously-used one.

Keywords: planning, scheduling, risk management, project duration, site safety

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Authors: E. Erdogan, M. Ulusoy

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In terms of tourism, the concept of sustainability can be defined as preserving and developing natural, historical, cultural, social, and aesthetic values and enabling their permanency. Sustainable tourism aims to preserve natural, historical, cultural, and social resources, also by supporting economic progress protecting economic development and environmental values that emerge as a consequence of tourism activities. Cultural tourism feeds on sustainable cultural treasures inherently and is the most effective touristic activity. Traditional configurations and structural characteristics play an important role in generating cultural tourism in a region. Sustainable cultural tourism is related to trips upon people who embark with the aim of visiting culturally rich regions, learning about and observing fast-disappearing lifestyles and collecting cultural values as memories. With its huge tourism potential, Karadağ is the most significant cultural asset of the Karaman province, possessing unique riches in terms of cultural world history. Host to one of the most important Byzantine cities in Anatolia, Karadağ is like an open-air museum with its unparalleled architectural structures. There is a village named Madenşehir in the plain at the outskirts of Karadağ, near to which are located the “Thousand and One Churches” ruins. The 80-household house is located near the ruins in an area that been declared a 1st degree historic preservation district. stones gathered from local churches were used in the construction of these households. A ministry has assigned a new residential site near the boundaries of the 2nd degree preservation district, and the decision has been made to move the occupants to this area. The most important issue here is to enable locals’ sociocultural and socioeconomic sustainability. It is also important to build these structures in a manner compatible with the historical visual look, ecological system and environmental awareness. Therefore this new site will be planned as touristic area in terms of sustainable cultural tourism and in these new plans, shall fulfill functions oriented toward both tourists and locals. It is very important that this change be sustainable and also support cultural tourism.

Keywords: cultural tourism, new village settlement, socio cultural sustainability, “thousand and one churches” site

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Authors: Leticia Leandro Rade, Amanda Silva de Sousa, Suman Das, Wesley Generoso, Mayara Chagas Ávila, Plinio Salmazo Vieira, Antonio Bonomi, Gabriela Persinoti, Mario Tyago Murakami, Thomas Michael Makris, Leticia Maria Zanphorlin

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Alkenes have an economic appeal, especially in the biofuels field, since they are precursors for drop-in biofuels production, which have similar chemical and physical properties to the conventional fossil fuels, with no oxygen in their composition. After the discovery of the first P450 CYP152 OleTJE in 2011, reported with its unique property of decarboxylating fatty acids (FA), by using hydrogen peroxide as a cofactor and producing 1-alkenes as the main product, the scientific and technological interest in this family of enzymes vastly increased. In this context, the present work presents a new decarboxylase (OleTRN) with low similarity with OleTJE (32%), its biochemical characterization, and structure elucidation. As main results, OleTRN presented a high yield of expression and purity, optimum reaction conditions at 35 °C and pH from 6.5 to 8.0, and higher specificity for oleic acid. Besides that, structure-guided mutations were performed and according to the functional characterizations, it was observed that some mutations presented different specificity and chemoselectivity by varying the chain-length of FA substrates from 12 to 20 carbons. These results are extremely interesting from a biotechnological perspective as those characteristics could diversify the applications and contribute to designing better cytochrome P450 decarboxylases. Considering that peroxygenases have the potential activity of decarboxylating and hydroxylating fatty acids and that the elucidation of the intriguing mechanistic involved in the decarboxylation preferential from OleTJE is still a challenge, the elucidation of OleTRN structure and the functional characterizations of OleTRN and its mutants contribute to new information about CYP152. Besides that, the work also contributed to the discovery of a new decarboxylase with a different selectivity profile from OleTJE, which allows a wide range of applications.

Keywords: P450, decarboxylases, alkenes, biofuels

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Authors: Lintang Putra Sadewa, Ilham Prasetya Budhi

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Slope monitoring program has now become a mandatory campaign for any open pit mines around the world to operate safely. Utilizing various slope monitoring instruments and strategies, miners are now able to deliver precise decisions in mitigating the risk of slope failures which can be catastrophic. Currently, the most sophisticated slope monitoring technology available is the Slope Stability Radar (SSR), whichcan measure wall deformation in submillimeter accuracy. One of its eminent features is that SSRcan provide a timely warning by automatically raise an alarm when a predetermined rate-of-movement threshold is reached. However, establishing proper alarm thresholds is arguably one of the onerous challenges faced in any slope monitoring program. The difficulty mainly lies in the number of considerations that must be taken when generating a threshold becausean alarm must be effectivethat it should limit the occurrences of false alarms while alsobeing able to capture any real wall deformations. In this sense, experience shows that a site-specific alarm thresholdtendsto produce more reliable results because it considers site distinctive variables. This study will attempt to determinealarming thresholds for safety-critical monitoring based on an empirical model of slope deformation behaviour that is defined statistically fromdeformation data captured by the Slope Stability Radar (SSR). The study area comprises of upright-dipping highwall setting in a coal mining area with intense mining activities, andthe deformation data used for the study were recorded by the SSR throughout the year 2022. The model is site-specific in nature thus, valuable information extracted from the model (e.g., time-to-failure, onset-of-acceleration, and velocity) will be applicable in setting up site-specific alarm thresholds and will give a clear understanding of how deformation trends evolve over the area.

Keywords: safety-critical monitoring, alarming strategy, slope deformation behaviour model, coal mining

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Authors: Guilherme M. Pereira, Kimmo Teinïla, Danilo Custódio, Risto Hillamo, Célia Alves, Pérola de C. Vasconcellos

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South American large cities often present serious air pollution problems and their atmosphere composition is influenced by a variety of emissions sources. The South American Emissions Megacities, and Climate project (SAEMC) has focused on the study of emissions and its influence on climate in the South American largest cities and it also included Lima (Peru) and Medellin (Colombia), sites where few studies of the genre were done. Lima is a coastal city with more than 8 million inhabitants and the second largest city in South America. Medellin is a 2.5 million inhabitants city and second largest city in Colombia; it is situated in a valley. The samples were collected in quartz fiber filters in high volume samplers (Hi-Vol), in 24 hours of sampling. The samples were collected in intensive campaigns in both sites, in July, 2010. Several species were determined in the aerosol samples of Lima and Medellin. Organic and elemental carbon (OC and EC) in thermal-optical analysis; biomass burning tracers (levoglucosan - Lev, mannosan - Man and galactosan - Gal) in high-performance anion exchange ion chromatography with mass spectrometer detection; water soluble ions in ion chromatography. The average particulate matter was similar for both campaigns, the PM10 concentrations were above the recommended by World Health Organization (50 µg m⁻³ – daily limit) in 40% of the samples in Medellin, while in Lima it was above that value in 15% of the samples. The average total ions concentration was higher in Lima (17450 ng m⁻³ in Lima and 3816 ng m⁻³ in Medellin) and the average concentrations of sodium and chloride were higher in this site, these species also had better correlations (Pearson’s coefficient = 0,63); suggesting a higher influence of marine aerosol in the site due its location in the coast. Sulphate concentrations were also much higher at Lima site; which may be explained by a higher influence of marine originated sulphate. However, the OC, EC and monosaccharides average concentrations were higher at Medellin site; this may be due to the lower dispersion of pollutants due to the site’s location and a larger influence of biomass burning sources. The levoglucosan average concentration was 95 ng m⁻³ for Medellin and 16 ng m⁻³ and OC was well correlated with levoglucosan (Pearson’s coefficient = 0,86) in Medellin; suggesting a higher influence of biomass burning over the organic aerosol in this site. The Lev/Man ratio is often related to the type of biomass burned and was close to 18, similar to the observed in previous studies done at biomass burning impacted sites in the Amazon region; backward trajectories also suggested the transport of aerosol from that region. Biomass burning appears to have a larger influence on the air quality in Medellin, in addition the vehicular emissions; while Lima showed a larger influence of marine aerosol during the study period.

Keywords: aerosol transport, atmospheric particulate matter, biomass burning, SAEMC project

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Authors: Sathyaseelan Subramaniam, Jacqueline Bober, Jennifer Chao, Shahriar Zehtabchi

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Background: Emergency physicians frequently evaluate skin and soft tissue infections in order to differentiate abscess from cellulitis. This helps determine which patients will benefit from incision and drainage. Our objective was to determine the operating characteristics of point-of-care ultrasound (POCUS) compared to clinical examination in identifying abscesses in emergency department (ED) patients with features of skin and soft tissue infections. Methods: We performed a comprehensive search in the following databases: Medline, Web of Science, EMBASE, CINAHL and Cochrane Library. Trials were included if they compared the operating characteristics of POCUS with clinical examination in identifying skin and soft tissue abscesses. Trials that included patients with oropharyngeal abscesses or that requiring abscess drainage in the operating room were excluded. The presence of an abscess was determined by pus drainage. No pus seen on incision or resolution of symptoms without pus drainage at follow up, determined the absence of an abscess. Quality of included trials was assessed using GRADE criteria. Operating characteristics of POCUS are reported as sensitivity, specificity, positive likelihood (LR+) and negative likelihood (LR-) ratios and the respective 95% confidence intervals (CI). Summary measures were calculated by generating a hierarchical summary receiver operating characteristic model (HSROC). Results: Out of 3203 references identified, 5 observational studies with 615 patients in aggregate were included (2 adults and 3 pediatrics). We rated the quality of 3 trials as low and 2 as very low. The operating characteristics of POCUS and clinical examination in identifying soft tissue abscesses are presented in the table. The HSROC for POCUS revealed a sensitivity of 96% (95% CI = 89-98%), specificity of 79% (95% CI = 71-86), LR+ of 4.6 (95% CI = 3.2-6.8), and LR- of 0.06 (95% CI = 0.02-0.2). Conclusion: Existing evidence indicates that POCUS is useful in identifying abscesses in ED patients with skin or soft tissue infections.

Keywords: abscess, point-of-care ultrasound, pocus, skin and soft tissue infection

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Authors: Zulfiqar Ali, Umar Saleem, Muhammad Junaid, Rizwan Tahir

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Gulzar Mahal is a heritage site located in the city of Bahawalpur, Pakistan. The site is under a process of degradation, as cracks are appearing on the walls, roofs, and floor around the building due to differential settlement. To preserve the integrity of the structure, a geotechnical distress evaluation was carried out to evaluate the causal factors and recommend remediation measures. The research involved the characterization of the problematic soil and analysis of the observed distress with respect to the geotechnical properties. Both conventional lab and field tests were used in conjunction with the unconventional techniques like; Electrical Resistivity Tomography (ERT) and FEA. The temporal, geophysical and geotechnical evaluations have concluded that the foundation soil over the past was subjected to variations in the land use, poor drainage patterns, overloading and fluctuations in groundwater table all contributing to the differential settlements manifesting in the form of the visible shear crack across the length and breadth of the building.

Keywords: differential settlement, distress evaluation, finite element analysis, Gulzar Mahal

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Authors: Kuan Lun Pan, Moo Been Chang

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Volatile organic compounds (VOCs) are one of major air contaminants, and they can react with nitrogen oxides (NOx) in atmosphere to form ozone (O3) and peroxyacetyl nitrate (PAN) with solar irradiation, leading to environmental hazards. In addition, some VOCs are toxic at low concentration levels and cause adverse effects on human health. How to effectively reduce VOCs emission has become an important issue. Thermal catalysis is regarded as an effective way for VOCs removal because it provides oxidation route to successfully convert VOCs into carbon dioxide (CO2) and water (H2O(g)). Single perovskite-type catalysts are promising for VOC removal, and they are of good potential to replace noble metals due to good activity and high thermal stability. Single perovskites can be generally described as ABO3 or A2BO4, where A-site is often a rare earth element or an alkaline. Typically, the B-site is transition metal cation (Fe, Cu, Ni, Co, or Mn). Catalytic properties of perovskites mainly rely on nature, oxidation states and arrangement of B-site cation. Interestingly, single perovskites could be further synthesized to form double perovskite-type catalysts which can simply be represented by A2B’B”O6. Likewise, A-site stands for an alkaline metal or rare earth element, and the B′ and B′′ are transition metals. Double perovskites possess unique surface properties. In structure, three-dimensional of B-site with ordered arrangement of B’O6 and B”O6 is presented alternately, and they corner-share octahedral along three directions of the crystal lattice, while cations of A-site position between the void of octahedral. It has attracted considerable attention due to specific arrangement of alternating B-site structure. Therefore, double perovskites may have more variations than single perovskites, and this greater variation may promote catalytic performance. It is expected that activity of double perovskites is higher than that of single perovskites toward VOC removal. In this study, double perovskite-type catalyst (La2CoMnO6) is prepared and evaluated for VOC removal. Also, single perovskites including LaCoO3 and LaMnO3 are tested for the comparison purpose. Toluene (C7H8) is one of the important VOCs which are commonly applied in chemical processes. In addition to its wide application, C7H8 has high toxicity at a low concentration. Therefore, C7H8 is selected as the target compound in this study. Experimental results indicate that double perovskite (La2CoMnO6) has better activity if compared with single perovskites. Especially, C7H8 can be completely oxidized to CO2 at 300oC as La2CoMnO6 is applied. Characterization of catalysts indicates that double perovskite has unique surface properties and is of higher amounts of lattice oxygen, leading to higher activity. For durability test, La2CoMnO6 maintains high C7H8 removal efficiency of 100% at 300oC and 30,000 h-1, and it also shows good resistance to CO2 (5%) and H2O(g) (5%) of gas streams tested. For various VOCs including isopropyl alcohol (C3H8O), ethanal (C2H4O), and ethylene (C2H4) tested, as high as 100% efficiency could be achieved with double perovskite-type catalyst operated at 300℃, indicating that double perovskites are promising catalysts for VOCs removal, and possible mechanisms will be elucidated in this paper.

Keywords: volatile organic compounds, Toluene (C7H8), double perovskite-type catalyst, catalysis

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Authors: Anyimc, C. J. Aneke, J. O. Orji, O. Nworie, U. C. C. Egbule

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The microbial examination and antimicrobial susceptibility profile of microorganism isolated from the salt mining site in Ebonyi state were evaluated in the present study using a standard microbiological technique. A total of 300 samples were randomly collected in three sample groups (A, B, and C) of 100 each. Isolation, Identification and characterization of organization present on the soil samples were determined by culturing, gram-staining and biochemical technique. The result showed the following organisms were isolated with their frequency as follow: Bacillus species (37.3%) and Staphylococcus species(23.5%) had the highest frequency in the whole Sample group A and B while Klebsiella specie (15.7%), Pseudomonas species(13.7%), and Erwinia species (9.8%) had the least. Rhizopus species (42.0%) and Aspergillus species (26.0%) were the highest fungi isolated, followed by Penicillum species (20.0%) while Mucor species (4.0%), and Fusarium species (8.0%) recorded the least. Sample group C showed high microbial population of all the microbial isolates when compared to sample group A and B. Disc diffusion method was used to determine the susceptibility of isolated bacteria to various antibiotics (oxfloxacin, pefloxacin, ciprorex, augumentin, gentamycin, ciproflox, septrin, ampicillin), while agar well diffusion method was used to determine the susceptibility of isolated fungi to some antifungal drugs (metronidazole, ketoconazole, itraconazole fluconazole). The antibacterial activity of the antibiotics used showed that ciproflux has the best inhibitory effect on all the test bacteria. Ketoconazole showed the highest inhibitory effect on the fungal isolates, followed by itraconazole, while metronidazole and fluconazole showed the least inhibitory effect on the entire test fungal isolates. Hence, the multiple drug resistance of most isolates to appropriate drugs of choice are of great public health concern and cells for periodic monitoring of antibiograms to detect possible changing patterns. Microbes isolated in the salt mining site can also be used as a source of gene(s) that can increase salt tolerance in different crop species through genetic engineering.

Keywords: microorganisms, antibacterial, antifungal, resistance, salt mining site, Ebonyi State

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Authors: Pia Wetzl

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The learning process of students can be optimized by combining online teaching with face-to-face sessions. So-called blended learning offers extensive flexibility as well as contact opportunities with fellow students and teachers. Furthermore, learning can be individualized and self-regulated. The aim of this article is to investigate which factors are necessary for blended learning to be successful. Semi-structured interviews were conducted with students (N = 60) and lecturers (N = 21) from different disciplines at two German universities. The questions focused on the perception of online, face-to-face and blended learning courses. In addition, questions focused on possible optimization potential and obstacles to practical implementation. The results show that on-site presence is very important for blended learning to be successful. If students do not get to know each other on-site, there is a risk of loneliness during the self-learning phases. This has a negative impact on motivation. From the perspective of the lecturers, the willingness of the students to participate in the sessions on-site is low. Especially when there is no obligation to attend, group work is difficult to implement because the number of students attending is too low. Lecturers would like to see more opportunities from the university and its administration to enforce attendance. In their view, this is the only way to ensure the success of blended learning. In addition, they see the conception of blended learning courses as requiring a great deal of time, which they are not always willing to invest. More incentives are necessary to keep the lecturers motivated to develop engaging teaching material. The study identifies factors that can help teachers conceptualize blended learning. It also provides specific implementation advice and identifies potential impacts. This catalogue has great value for the future-oriented development of courses at universities. Future studies could test its practical use.

Keywords: blended learning, higher education, teachers, student learning, qualitative research

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Authors: Collins C. Chiemeke, I. B. Osazuwa, S. O. Ibe, G. N. Egwuonwu, C. D. Ani, E. C. Chii

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The study area is Zaria, located in the basement complex of northern Nigeria. The rock type forming the major part of the Zaria batholith is granite. This research work was carried out to compare the responses of seismic refraction tomography and resistivity tomography in the same geologic environment and under the same conditions. Hence, the choice of the site that has a visible granitic outcrop that extends across a narrow stream channel and is flanked by unconsolidated overburden, a neutral profile that was covered by plain overburden and a site with thick lateritic cover became necessary. The results of the seismic and resistivity tomography models reveals that seismic velocity and resistivity does not always simultaneously increase with depth, but their responses in any geologic environment are determined by changes in the mechanical and chemical content of the rock types rather than depth.

Keywords: environment, resistivity, response, seismic, velocity

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Authors: Ishwor Paudel, Pratima Gautam, Sandeep Bhattarai

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Bile spillage occurs frequently during laparoscopic cholecystectomy yet its impact on postoperative outcomes remains unknown. It might not be as innocuous as some surgeons tend to believe and in fact, might be associated with post-operative surgical site infections (SSI). It often leads to patient dissatisfaction, emergency department visits, with subsequent readmission and additional procedures. While some authors found no increase in SSI following bile spillage (BS) compared to non-bile spillage cases, others found bile spillage to be associated with SSI. Therefore we sought to examine whether bile spillage is indeed associated with an increased risk of postoperative wound infections after laparoscopic cholecystectomy. I hypothesize that patients who experience BS during operation, have an increased risk of SSI compared to those who do not. This is a prospective observational study conducted in the Department of Surgery, Patan Hospital over a period of one year. Patients undergoing Laparoscopic cholecystectomy were included and bile spillage, if happened was noted. All cases were followed up for 30 days and SSI was diagnosed as per CDC-defined criteria. Fisher’s test was applied to compare SSI in bile spillage versus non-bile spillage cases. A total of 112 patients were included in the final analysis. Bile spillage occurred in 20 cases and was absent in the rest i.e.92 cases. Among bile spillage cases, SSI was found in 4 cases (20%), whereas in nonbile spillage cases, SSI was found in 8 cases (8.7%). However, it was statistically not significant (p-value>0.05). Eleven (92%) cases were superficial SSI and one was an organ-space infection. No mortality or 30-day readmission. Spillage of Gallbladder content does not lead to an increase in SSIs. However as the rate of SSI is still higher, Surgeons should be careful to avoid iatrogenic gallbladder perforation and in case of bile spillage thorough peritoneal irrigation with normal saline should be done.

Keywords: organ space infection, Laparoscopic cholecystectomy, biliary spillage, surgical site infection

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Authors: Bliss Singhal

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Epilepsy is a prevalent neurological disorder affecting approximately 50 million individuals worldwide and 1.2 million Americans. There exist millions of pediatric patients with intractable epilepsy, a condition in which seizures fail to come under control. The occurrence of seizures can result in physical injury, disorientation, unconsciousness, and additional symptoms that could impede children's ability to participate in everyday tasks. Predicting seizures can help parents and healthcare providers take precautions, prevent risky situations, and mentally prepare children to minimize anxiety and nervousness associated with the uncertainty of a seizure. This research proposes a comprehensive framework to predict seizures in pediatric patients by evaluating machine learning algorithms on unimodal neuroimaging data consisting of electroencephalogram signals. The bandpass filtering and independent component analysis proved to be effective in reducing the noise and artifacts from the dataset. Various machine learning algorithms’ performance is evaluated on important metrics such as accuracy, precision, specificity, sensitivity, F1 score and MCC. The results show that the deep learning algorithms are more successful in predicting seizures than logistic Regression, and k nearest neighbors. The recurrent neural network (RNN) gave the highest precision and F1 Score, long short-term memory (LSTM) outperformed RNN in accuracy and convolutional neural network (CNN) resulted in the highest Specificity. This research has significant implications for healthcare providers in proactively managing seizure occurrence in pediatric patients, potentially transforming clinical practices, and improving pediatric care.

Keywords: intractable epilepsy, seizure, deep learning, prediction, electroencephalogram channels

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Authors: D. Sigirli

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The Receiver Operating Characteristic (ROC) curve is a commonly used statistical tool for evaluating the diagnostic performance of screening and diagnostic test with continuous or ordinal scale results which aims to predict the presence or absence probability of a condition, usually a disease. When the test results were measured as numeric values, sensitivity and specificity can be computed across all possible threshold values which discriminate the subjects as diseased and non-diseased. There are infinite numbers of possible decision thresholds along the continuum of the test results. The ROC curve presents the trade-off between sensitivity and the 1-specificity as the threshold changes. The empirical ROC curve which is a non-parametric estimator of the ROC curve is robust and it represents data accurately. However, especially for small sample sizes, it has a problem of variability and as it is a step function there can be different false positive rates for a true positive rate value and vice versa. Besides, the estimated ROC curve being in a jagged form, since the true ROC curve is a smooth curve, it underestimates the true ROC curve. Since the true ROC curve is assumed to be smooth, several smoothing methods have been explored to smooth a ROC curve. These include using kernel estimates, using log-concave densities, to fit parameters for the specified density function to the data with the maximum-likelihood fitting of univariate distributions or to create a probability distribution by fitting the specified distribution to the data nd using smooth versions of the empirical distribution functions. In the present paper, we aimed to propose a smooth ROC curve estimation based on the boundary corrected kernel function and to compare the performances of ROC curve smoothing methods for the diagnostic test results coming from different distributions in different sample sizes. We performed simulation study to compare the performances of different methods for different scenarios with 1000 repetitions. It is seen that the performance of the proposed method was typically better than that of the empirical ROC curve and only slightly worse compared to the binormal model when in fact the underlying samples were generated from the normal distribution.

Keywords: empirical estimator, kernel function, smoothing, receiver operating characteristic curve

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Authors: Opal Giulianelli, Pegah Zamani

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Sustainable food has become a frequently debated topic in recent years due to a consumer push for environmentally sustainable food. While some research works on improving the monoculture farm systems that are currently in use, others focus on expanding the definition of sustainable food systems. This research looks at those concepts of alternative food systems applied to a more extensive city system. The goal is to create a theoretical site plan that could be implemented in emerging cities and other urban environments. This site plan combines the ideas of environmentally sustainable food development, such as food forests, urban farming, and community gardens. This would represent one part of a larger sustainable food system that can be altered depending on the environment or the people it is serving. However, this research is being carried out with the southeast United States in mind and, therefore, may prove difficult to apply to other regions, especially those of radically different climates.

Keywords: alternative food systems, urban design, food forests, aquaponics, hydroponics, food security, food system design

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Authors: Ayesha Sanjana Kawser Parsha

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S-acylation is an irreversible bond in which cysteine residues are linked to fatty acids palmitate (74%) or stearate (22%), either at the COOH or NH2 terminal, via a thioester linkage. There are several experimental methods that can be used to identify the S-palmitoylation site; however, since they require a lot of time, computational methods are becoming increasingly necessary. There aren't many predictors, however, that can locate S- palmitoylation sites in Arabidopsis Thaliana with sufficient accuracy. This research is based on the importance of building a better prediction tool. To identify the type of machine learning algorithm that predicts this site more accurately for the experimental dataset, several prediction tools were examined in this research, including the GPS PALM 6.0, pCysMod, GPS LIPID 1.0, CSS PALM 4.0, and NBA PALM. These analyses were conducted by constructing the receiver operating characteristics plot and the area under the curve score. An AI-driven deep learning-based prediction tool has been developed utilizing the analysis and three sequence-based input data, such as the amino acid composition, binary encoding profile, and autocorrelation features. The model was developed using five layers, two activation functions, associated parameters, and hyperparameters. The model was built using various combinations of features, and after training and validation, it performed better when all the features were present while using the experimental dataset for 8 and 10-fold cross-validations. While testing the model with unseen and new data, such as the GPS PALM 6.0 plant and pCysMod mouse, the model performed better, and the area under the curve score was near 1. It can be demonstrated that this model outperforms the prior tools in predicting the S- palmitoylation site in the experimental data set by comparing the area under curve score of 10-fold cross-validation of the new model with the established tools' area under curve score with their respective training sets. The objective of this study is to develop a prediction tool for Arabidopsis Thaliana that is more accurate than current tools, as measured by the area under the curve score. Plant food production and immunological treatment targets can both be managed by utilizing this method to forecast S- palmitoylation sites.

Keywords: S- palmitoylation, ROC PLOT, area under the curve, cross- validation score

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Authors: Avani Saraswat

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The paper studies the present condition of Taj Mahal (the UNESCO world heritage site) and the reasons behind deterioration. Analysis is done to explore the reasons behind this building to be included in the list of adopt heritage scheme, by the Government of India. The aim is to find out the future effects on Taj Mahal after being adopted by a private body. Finally, it suggests solutions which can lead to improvement of the present condition of the building. In order to establish a research, a further analysis is done through a case study of Red Fort, New Delhi (another UNESCO world heritage site). This monument was given to Dalmia Group of India Pvt. Ltd. for the tenure of 5 years. Paper discusses the consequences of privatization on Red Fort and then analyze it for Taj Mahal. It terms monument as riches of a heritage chest, not as a commercial tourist place. The study is concluded with the ideas and suggestions proposed for saving Taj Mahal and advantages on improving the health of the building.

Keywords: privatisation of heritage, heritage, Taj Mahal, adopt heritage scheme

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Authors: Taghi-Zada T. G., Hajiyeva U. K.

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Recent years, due to the development of tourism, migration and globalization, brucellosis has spread to non-endemic regions of the country in Azerbaijan and this disease has become one of the main priority areas of medicine. In our daily practice, we face patients with specific symptoms of brucellosis and also infected with this disease but misdiagnosed. It should also be noted that the symptoms and signs of brucellosis are very diverse, and since none of these signs are specific enough to confirm the diagnosis, it creates difficulties in its timely detection and diagnosis. The main purpose of the work. Therefore, the main goal of the work is to investigate the cases of delay in making the correct diagnosis in children with brucellosis and the mistakes in this matter. Material and method. 50 children with brucellosis between the ages of 6 months and 17 years were examined. The medical history and anamnesis of these children were collected, clinical-instrumental examination, and serological tests for brucellosis were performed. Patients were divided into 2 groups, taking into account the specificity of symptoms and the timely diagnosis Results. Group I included 15 (40%) children aged 3-17 years. The main specific symptoms of brucellosis in these patients; persistent or long-term fever, night sweats, arthralgia were observed. In addition to specific symptoms, anamnesis and a specific serological test confirmed the diagnosis of brucellosis. 30 (60%) patients included in group II were misdiagnosed. 3 patients (up to 1 year) were diagnosed with sepsis, 6 with acute rheumatic fever, 10 with systemic diseases, 2 with tuberculosis, 5 with Covid 19, and 4 with unspecified fever. However, we included serological tests. detailed examination revealed the presence of brucellosis in them. As can be seen, compared to group I (40%) children included in group II (60%) In modern times, brucellosis manifests itself with its own characteristics, that is, imitating a number of other diseases, which has led to wrong diagnosis. Conclusion. Thus, the lack of specificity of clinical symptoms during brucellosis in children makes diagnosis difficult, causes mistakes and non-recognition of the disease. With this in mind, physicians in predominantly endemic and even sub-endemic areas should remain vigilant about this disease and consider brucellosis in the differential diagnosis of almost every unexplained medical problem until proven otherwise.

Keywords: brucellosis, pediatrics, diagnostics, serological tests

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Authors: Muzaffar Shaikh, Shoaib Shaikh, Mark Moyou, Gaby Hawat

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This paper presents the use of Analytic Hierarchy Process (AHP) in evaluating the site selection of a new plant by a corporation. Due to intense competition at a global level, multinational corporations are continuously striving to minimize production and shipping costs of their products. One key factor that plays significant role in cost minimization is where the production plant is located. In the U.S. for example, labor and land costs continue to be very high while they are much cheaper in countries such as India, China, Indonesia, etc. This is why many multinational U.S. corporations (e.g. General Electric, Caterpillar Inc., Ford, General Motors, etc.), have shifted their manufacturing plants outside. The continued expansion of the Internet and its availability along with technological advances in computer hardware and software all around the globe have facilitated U.S. corporations to expand abroad as they seek to reduce production cost. In particular, management of multinational corporations is constantly engaged in concentrating on countries at a broad level, or cities within specific countries where certain or all parts of their end products or the end products themselves can be manufactured cheaper than in the U.S. AHP is based on preference ratings of a specific decision maker who can be the Chief Operating Officer of a company or his/her designated data analytics engineer. It serves as a tool to first evaluate the plant site selection criteria and second, alternate plant sites themselves against these criteria in a systematic manner. Examples of site selection criteria are: Transportation Modes, Taxes, Energy Modes, Labor Force Availability, Labor Rates, Raw Material Availability, Political Stability, Land Costs, etc. As a necessary first step under AHP, evaluation criteria and alternate plant site countries are identified. Depending upon the fidelity of analysis, specific cities within a country can also be chosen as alternative facility locations. AHP experience in this type of analysis indicates that the initial analysis can be performed at the Country-level. Once a specific country is chosen via AHP, secondary analyses can be performed by selecting specific cities or counties within a country. AHP analysis is usually based on preferred ratings of a decision-maker (e.g., 1 to 5, 1 to 7, or 1 to 9, etc., where 1 means least preferred and a 5 means most preferred). The decision-maker assigns preferred ratings first, criterion vs. criterion and creates a Criteria Matrix. Next, he/she assigns preference ratings by alternative vs. alternative against each criterion. Once this data is collected, AHP is applied to first get the rank-ordering of criteria. Next, rank-ordering of alternatives is done against each criterion resulting in an Alternative Matrix. Finally, overall rank ordering of alternative facility locations is obtained by matrix multiplication of Alternative Matrix and Criteria Matrix. The most practical aspect of AHP is the ‘what if’ analysis that the decision-maker can conduct after the initial results to provide valuable sensitivity information of specific criteria to other criteria and alternatives.

Keywords: analytic hierarchy process, multinational corporations, plant site selection, preference ratings

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Authors: Li Shiyue

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In an urban built environment, urban green space is scarce, especially around old residential areas. Due to the innate design deficiency and the non-core location of these areas, they lack green space, and the recreational opportunities of the surrounding residents are not guaranteed. Micro greenspace becomes a "patch" to compensate for the urban function. To realize the renewal and transformation of micro greenspace, and make it meet the use needs of most groups, this paper introduces the concept of inclusive design. Based on relevant research at home and abroad, this paper discusses the connotation and current situation of micro greenspace. Combining with the realistic conditions of China, this paper thinks about the planning path of inclusive renewal from the aspects of selecting micro greenspace transformation potential points and exploring the key points of site renewal. Among them, the key points of site renewal are explored from five angles: land guarantee, systematic coordination, refined design, and shared space creation, to provide useful references for related research and practice.

Keywords: inclusive design, micro greenspace, old city area, space renewal

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Authors: Nitin Dwivedi, Jigna Shah

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Brain tumor is metastatic neoplasm of central nervous system, in most of cases it is life threatening disease with low survival rate. Despite of enormous efforts in the development of therapeutics and diagnostic tools, the treatment of brain tumors and gliomas remain a considerable challenge in the area of neuro-oncology. The most reason behind of this the presence of physiological barriers including blood brain barrier and blood brain tumor barrier, lead to insufficient reach ability of therapeutic agents at the site of tumor, result of inadequate destruction of gliomas. So there is an indeed need empowerment of brain tumor imaging for better characterization and delineation of tumors, visualization of malignant tissue during surgery, and tracking of response to chemotherapy and radiotherapy. Multifunctional different generations of dendrimer offer an improved effort for potentiate drug delivery at the site of brain tumor and gliomas. So this article emphasizes the innovative dendrimer approaches in tumor targeting, tumor imaging and delivery of therapeutic agent.

Keywords: blood brain barrier, dendrimer, gliomas, nanotechnology

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Authors: Roopam Gaur, K. Chandramani Singh, Radhapiyari Laishram

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In order to produce lead free piezoceramics with optimum piezoelectric and dielectric properties, KNN modified with Li+ (as an A site dopant) and Sb5+ (as a B site dopant) (K0.49Na0.49Li0.02) (Nb0.96Sb0.04) O3 (referred as KNLNS in this paper) have been synthesized using solid state reaction method and conventional sintering technique. The ceramics were sintered in the narrow range of 10500C-10900C for 2-3 hours to get precise information about sintering parameters. Detailed study of dependence of microstructural, dielectric and piezoelectric properties on sintering conditions was then carried out. The study suggests that the volatility of the highly hygroscopic KNN ceramics is not only sensitive to sintering temperatures but also to sintering durations. By merely reducing the sintering duration for a given sintering temperature we saw an increase in the density of the samples which was supported by the increase in dielectric constants of the ceramics. And since density directly or indirectly affects almost all the associated properties, other dielectric and piezoelectric properties were also enhanced as we approached towards the most suitable sintering temperature and duration combination.

Keywords: piezoelectric, dielectric, Li, Sb, KNN, conventional sintering

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Authors: Laura Kupers, Julie Piérard, Niki Cauberg

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The use of exposed concrete (sometimes referred to as architectural concrete), keeps gaining popularity. Exposed concrete has the advantage to combine the structural properties of concrete with an aesthetic finish. However, for a successful aesthetic finish, much attention needs to be paid to the execution (formwork, release agent, curing, weather conditions…), the concrete composition (choice of the raw materials and mix proportions) as well as to its fresh properties. For the latter, a simple on-site pass/fail test could halt the casting of concrete not suitable for architectural concrete and thus avoid expensive repairs later. When architects opt for an exposed concrete, they usually want a smooth, uniform and nearly blemish-free surface. For this choice, a standard ‘construction’ concrete does not suffice. An aesthetic surface finishing requires the concrete to contain a minimum content of fines to minimize the risk of segregation and to allow complete filling of more complex shaped formworks. The concrete may neither be too viscous as this makes it more difficult to compact and it increases the risk of blow holes blemishing the surface. On the other hand, too much bleeding may cause color differences on the concrete surface. An easy pass/fail test, which can be performed on the site just before the casting, could avoid these problems. In case the fresh concrete fails the test, the concrete can be rejected. Only in case the fresh concrete passes the test, the concrete would be cast. The pass/fail tests are intended for a concrete with a consistency class S4. Five tests were selected as possible onsite pass/fail test. Two of these tests already exist: the K-slump test (ASTM C1362) and the Bauer Filter Press Test. The remaining three tests were developed by the BBRI in order to test the segregation resistance of fresh concrete on site: the ‘dynamic sieve stability test’, the ‘inverted cone test’ and an adapted ‘visual stability index’ (VSI) for the slump and flow test. These tests were inspired by existing tests for self-compacting concrete, for which the segregation resistance is of great importance. The suitability of the fresh concrete mixtures was also tested by means of a laboratory reference test (resistance to segregation) and by visual inspection (blow holes, structure…) of small test walls. More than fifteen concrete mixtures of different quality were tested. The results of the pass/fail tests were compared with the results of this laboratory reference test and the test walls. The preliminary laboratory results indicate that concrete mixtures ‘suitable’ for placing as exposed concrete (containing sufficient fines, a balanced grading curve etc.) can be distinguished from ‘inferior’ concrete mixtures. Additional laboratory tests, as well as tests on site, will be conducted to confirm these preliminary results and to set appropriate pass/fail values.

Keywords: exposed concrete, testing fresh concrete, segregation resistance, bleeding, consistency

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Authors: Lan Luo, Jingjing Huang, Huafang Li

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Introduction: Distressing low sexual desire is common in China, while the lack of reliable and valid instruments to evaluate symptoms of hypoactive sexual desire disorder (HSDD) impedes related research and clinical services. Aim: This study aimed to validate the reliability and validity of the Female Sexual Function Index (FSFI) and the Female Sexual Distress Scale-Desire/Arousal/Orgasm (FSDS-DAO) in Chinese female HSDD patients. Methods: We administered FSFI and FSDS-DAO in a convenient sample of Chinese adult women. Participants were diagnosed by a psychiatrist according to the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, Text Revision (DSM-IV-TR). Results: We had a valid analysis sample of 279 Chinese women, of which 107 were HSDD patients. The Cronbach's α of FSFI and FSDS-DAO were 0.947 and 0.956, respectively, and the intraclass correlation coefficients of which were 0.86 and 0.89, respectively (the interval was 13-15 days). The correlation coefficient between the Revised Adult Attachment Scale (RAAS) and FSFI (or FSDS-DAO) did not exceed 0.4; the area under the receiver operating characteristic (ROC) curve was 0. 83 when combined FSFI-d (the desire domain of FSFI) and FSDS-DAO to diagnose HSDD, which was significantly different from that of using these scales individually. FSFI-d of less than 2.7 (1.2-6) and FSDS-DAO of no less than 15 (0-60) (Sensitivity 65%, Specificity 83%), or FSFI-d of no more than 3.0 (1.2-6) and FSDS-DAO of no less than 14 (0-60) (Sensitivity 74%, Specificity 77%) can be used as cutoff scores in clinical research or outpatient screening. Clinical implications: FSFI (including FSFI-d) and FSDS-DAO are suitable for the screening and evaluation of Chinese female HSDD patients of childbearing age. Strengths and limitations: Strengths include a thorough validation of FSFI and FSDS-DAO and the exploration of the cutoff score combing FSFI-d and FSDS-DAO. Limitations include a small convenience sample and the requirement of being sexually active for HSDD patients. Conclusion: FSFI (including FSFI-d) and FSDS-DAO have good internal consistency, test-retest reliability, construct validity, and criterion validity in Chinese female HSDD patients of childbearing age.

Keywords: sexual desire, sexual distress, hypoactive sexual desire disorder, scale

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Authors: Yu-Min Chang, Kuo-Sheng Tsai, Hung-Te Tsai, Chia-Hsin Li

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This paper presents the early-warning lights classification management system for industrial parks promoted by the Taiwan Environmental Protection Administration (EPA) since 2011, including the definition of each early-warning light, objectives, action program and accomplishments. All of the 151 industrial parks in Taiwan were classified into four early-warning lights, including red, orange, yellow and green, for carrying out respective pollution management according to the monitoring data of soil and groundwater quality, regulatory compliance, and regulatory listing of control site or remediation site. The Taiwan EPA set up a priority list for high potential polluted industrial parks and investigated their soil and groundwater qualities based on the results of the light classification and pollution potential assessment. In 2011-2013, there were 44 industrial parks selected and carried out different investigation, such as the early warning groundwater well networks establishment and pollution investigation/verification for the red and orange-light industrial parks and the environmental background survey for the yellow-light industrial parks. Among them, 22 industrial parks were newly or continuously confirmed that the concentrations of pollutants exceeded those in soil or groundwater pollution control standards. Thus, the further investigation, groundwater use restriction, listing of pollution control site or remediation site, and pollutant isolation measures were implemented by the local environmental protection and industry competent authorities; the early warning lights of those industrial parks were proposed to adjust up to orange or red-light. Up to the present, the preliminary positive effect of the soil and groundwater quality management system for industrial parks has been noticed in several aspects, such as environmental background information collection, early warning of pollution risk, pollution investigation and control, information integration and application, and inter-agency collaboration. Finally, the work and goal of self-initiated quality management of industrial parks will be carried out on the basis of the inter-agency collaboration by the classified lights system of early warning and management as well as the regular announcement of the status of each industrial park.

Keywords: industrial park, soil and groundwater quality management, early-warning lights classification, SOP for reporting and treatment of monitored abnormal events

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

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

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

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