Search results for: site specificity

Authors: Marzia Bilkiss, Muhammad J. A. Shiddiky, Mostafa K. Masud, Prabhakaran Sambasivam, Ido Bar, Jeremy Brownlie, Rebecca Ford

Abstract:

A greater achievement in the Integrated Disease Management (IDM) to prevent the loss would result from early diagnosis and quantitation of the causal pathogen species for accurate and timely disease control. This could significantly reduce costs to the growers and reduce any flow on impacts to the environment from excessive chemical spraying. Necrotrophic fungal disease botrytis grey mould, caused by Botrytis cinerea and Botrytis fabae, significantly reduce temperate legume yield and grain quality during favourable environmental condition in Australia and worldwide. Several immunogenic and molecular probe-type protocols have been developed for their diagnosis, but these have varying levels of species-specificity, sensitivity, and consequent usefulness within the paddock. To substantially improve speed, accuracy, and sensitivity, advanced nanoparticle-based biosensor approaches have been developed. For this, two sets of primers were designed for both Botrytis cinerea and Botrytis fabae which have shown the species specificity with initial sensitivity of two genomic copies/µl in pure fungal backgrounds using multiplexed quantitative PCR. During further validation, quantitative PCR detected 100 spores on artificially infected legume leaves. Simultaneously an electro-catalytic assay was developed for both target fungal DNA using functionalised magnetic nanoparticles. This was extremely sensitive, able to detect a single spore within a raw total plant nucleic acid extract background. We believe that the translation of this technology to the field will enable quantitative assessment of pathogen load for future accurate decision support of informed botrytis grey mould management.

Keywords: biosensor, botrytis grey mould, sensitive, species specific

Procedia PDF Downloads 152

Authors: Ayesha S. Rahman, Khaled Haddad, Ataur Rahman

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At-site flood frequency analysis is used to estimate flood quantiles when at-site record length is reasonably long. In Australia, FLIKE software has been introduced for at-site flood frequency analysis. The advantage of FLIKE is that, for a given application, the user can compare a number of most commonly adopted probability distributions and parameter estimation methods relatively quickly using a windows interface. The new version of FLIKE has been incorporated with the multiple Grubbs and Beck test which can identify multiple numbers of potentially influential low flows. This paper presents a case study considering six catchments in eastern Australia which compares two outlier identification tests (original Grubbs and Beck test and multiple Grubbs and Beck test) and two commonly applied probability distributions (Generalized Extreme Value (GEV) and Log Pearson type 3 (LP3)) using FLIKE software. It has been found that the multiple Grubbs and Beck test when used with LP3 distribution provides more accurate flood quantile estimates than when LP3 distribution is used with the original Grubbs and Beck test. Between these two methods, the differences in flood quantile estimates have been found to be up to 61% for the six study catchments. It has also been found that GEV distribution (with L moments) and LP3 distribution with the multiple Grubbs and Beck test provide quite similar results in most of the cases; however, a difference up to 38% has been noted for flood quantiles for annual exceedance probability (AEP) of 1 in 100 for one catchment. These findings need to be confirmed with a greater number of stations across other Australian states.

Keywords: floods, FLIKE, probability distributions, flood frequency, outlier

Procedia PDF Downloads 417

Authors: Silvia Di Turi, Nicolandrea Calabrese, Francesca Caffari, Giulia Centi, Francesca Margiotta, Giovanni Murano, Laura Ronchetti, Paolo Signoretti, Lisa Volpe, Domenico Palladino

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Recent European policies have been set ambitious targets aimed at significantly reducing CO2 emissions by 2030, with a long-term vision of transforming existing buildings into Zero-Emissions Buildings (ZEmB) by 2050. This vision represents a key point for the energy transition as the whole building stock currently accounts for 36% of total energy consumption across the Europe, mainly due to their poor energy performance. The challenge towards Zero-Emissions Buildings is particularly felt in Italy, where a significant number of buildings with historical significance or situated within protected/constrained areas can be found. Furthermore, an estimated 70% of the national building stock are built before 1976, indicating a widespread issue of poor energy performance. Addressing the energy ineƯiciency of these buildings is crucial to refining a comprehensive energy renovation approach aimed at facilitating their energy transition. In this framework the current study focuses on analysing a challenging complex of buildings to be totally restored through significant energy renovation interventions. The goal is to recover these disused buildings situated in a significant archaeological zone of Rome, contributing to the restoration and reintegration of this historically valuable site, while also oƯering insights useful for achieving zeroemission requirements for buildings within such contexts. In pursuit of meeting the stringent zero-emission requirements, a comprehensive study was carried out to assess the complex of buildings, envisioning substantial renovation measures on building envelope and plant systems and incorporating renewable energy system solutions, always respecting and preserving the historic site. An energy audit of the complex of buildings was performed to define the actual energy consumption for each energy service by adopting the hourly calculation methods. Subsequently, significant energy renovation interventions on both building envelope and mechanical systems have been examined respecting the historical value and preservation of site. These retrofit strategies have been investigated with threefold aims: 1) to recover the existing buildings ensuring the energy eƯiciency of the whole complex of buildings, 2) to explore which solutions have allowed achieving and facilitating the ZEmB status, 3) to balance the energy transition requirements with the sustainable aspect in order to preserve the historic value of the buildings and site. This study has pointed out the potentiality and the technical challenges associated with implementing renovation solutions for such buildings, representing one of the first attempt towards realizing this ambitious target for this type of building.

Keywords: energy conservation and transition, complex of buildings in historic site, zero-emission buildings, energy efficiency recovery

Procedia PDF Downloads 38

Authors: M. Bučas, A. Skersonas, E. Ivanauskas, J. Lesutienė, N. Nika, G. Srėbalienė, E. Tiškus, J. Gintauskas, A.Šaškov, G. Martin

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Submerged vegetation enhances heterogeneity of sublittoral habitats; therefore, macrophyte stands are essential elements of aquatic ecosystems to maintain a diverse fish fauna. Fish-habitat relations have been extensively studied in streams and coastal waters, but in lakes and estuaries are still underestimated. The aim of this study is to assess temporal (diurnal and seasonal) patterns of fish juvenile assemblages associated with common submerged macrophyte habitats, which have significantly spread during the recent decade in the upper littoral part of the Curonian Lagoon. The assessment was performed by means of an upgraded pop-up net approach resulting in much precise sampling versus other techniques. The optimal number of samples (i.e., pop-up nets) required to cover>80% of the total number of fish species depended on the time of the day in both study sites: at least 7and 9 nets in the evening (18-24 pm) in the Southern and Northern study sites, respectively. In total, 14 fish species were recorded, where perch and roach dominated (respectively 48% and 24%). From multivariate analysis, water salinity and seasonality (temperature or sampling month) were primary factors determining fish assemblage composition. The southern littoral area, less affected by brackish water conditions, hosted a higher number of species (13) than in the Northern site (8). In the latter site, brackish water tolerant species (three-spined and nine-spined sticklebacks, spiny loach, roach, and round goby) were more abundant than in the Southern site. Perch and ruffe dominated in the Southern site. Spiny loach and nine-spined stickleback were more frequent in September, while ruffe, perch, and roach occurred more in July. The diel dynamics of the common species such as perch, roach, and ruffe followed the general pattern, but it was species specific and depended on the study site, habitat, and month. The species composition between macrophyte habitats did not significantly differ; however, it differed from the results obtained in 2005 at both study sites indicating the importance of expanded charophyte stands during the last decade in the littoral zone.

Keywords: diel dynamics, charophytes, pondweeds, herbivorous and benthivorous fishes, littoral, nursery habitat, shelter

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Authors: Shalu Jain, Anju Bansal, Anup Kumar, Sunita Saxena

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Objectives: The novel TMPRSS2:ERG gene fusion is a common somatic event in prostate cancer that in some studies is linked with a more aggressive disease phenotype. Thus, this study aims to determine whether clinical variables are associated with the presence of TMPRSS2:ERG-fusion gene transcript in Indian patients of prostate cancer. Methods: We evaluated the clinical variables with presence and absence of TMPRSS2:ERG gene fusion in prostate cancer and BPH association of clinical patients. Patients referred for prostate biopsy because of abnormal DRE or/and elevated sPSA were enrolled for this prospective clinical study. TMPRSS2:ERG mRNA copies in samples were quantified using a Taqman chemistry by real time PCR assay in prostate biopsy samples (N=42). The T2:ERG assay detects the gene fusion mRNA isoform TMPRSS2 exon1 to ERG exon4. Results: Histopathology report has confirmed 25 cases as prostate cancer adenocarcinoma (PCa) and 17 patients as benign prostate hyperplasia (BPH). Out of 25 PCa cases, 16 (64%) were T2: ERG fusion positive. All 17 BPH controls were fusion negative. The T2:ERG fusion transcript was exclusively specific for prostate cancer as no case of BPH was detected having T2:ERG fusion, showing 100% specificity. The positive predictive value of fusion marker for prostate cancer is thus 100% and the negative predictive value is 65.3%. The T2:ERG fusion marker is significantly associated with clinical variables like no. of positive cores in prostate biopsy, Gleason score, serum PSA, perineural invasion, perivascular invasion and periprostatic fat involvement. Conclusions: Prostate cancer is a heterogeneous disease that may be defined by molecular subtypes such as the TMPRSS2:ERG fusion. In the present prospective study, the T2:ERG quantitative assay demonstrated high specificity for predicting biopsy outcome; sensitivity was similar to the prevalence of T2:ERG gene fusions in prostate tumors. These data suggest that further improvement in diagnostic accuracy could be achieved using a nomogram that combines T2:ERG with other markers and risk factors for prostate cancer.

Keywords: prostate cancer, genetic rearrangement, TMPRSS2:ERG fusion, clinical variables

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

Procedia PDF Downloads 122

Authors: Nathainail Bashir, Neil Anderson

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The objective of this study site was to investigate the current state of the practice with regards to karst detection methods and recommend the best method and pattern of arrays to acquire the desire results. Proper site investigation in karst prone regions is extremely valuable in determining the location of possible voids. Two geophysical techniques were employed: multichannel analysis of surface waves (MASW) and electric resistivity tomography (ERT).The MASW data was acquired at each test location using different array lengths and different array orientations (to increase the probability of getting interpretable data in karst terrain). The ERT data were acquired using a dipole-dipole array consisting of 168 electrodes. The MASW data was interpreted (re: estimated depth to physical top of rock) and used to constrain and verify the interpretation of the ERT data. The ERT data indicates poorer quality MASW data were acquired in areas where there was significant local variation in the depth to top of rock.

Keywords: dipole-dipole, ERT, Karst terrains, MASW

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Authors: M. Amelsakhi, A. Sohrabi-Bidar, A. Shareghi

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One of the most important issues about the structural damages caused by earthquake is the evaluating of the spectral response of the site on which the construction is built. This fact has demonstrated during many earlier earthquakes and many researchers’ reports have concerned with it. According to these reports, features of the site materials and geometry of the ground surface are considered the main factors. This study concentrates on the specific form of topographies like hills. Assessing of spectral responses of different points on the hills and beside demonstrates considerable differences between 1D and 2D methods of geotechnical analyses. A general trend of amplifications on the top of the hills and de-amplifications near the toe of the hills has been appeared within the acceleration, velocity and displacement response spectrums of horizontal motion. Evaluating of spectral responses of different sizes of the hills revealed that as much as the hill-size enlarges differences between spectral responses of 1D and 2D analyses transfers to longer range of periods and becomes wider.

Keywords: topography effect, amplification ratio, response spectrum, earth resources engineering

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Authors: Moizza Tahir, Uzma Bashir, Aisha Akhtar, Zainab Ansari, Sameen Ansari, Muhammad Ali Tahir

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Cancer burden has globally increased. Among cutaneous cancers basal cell carcinoma constitute vast majority of skin cancer. There is need for appropriate diagnostic, therapeutic and prognostic significance evaluation for skin cancers Present study report intraoperative frozen section (FS) histopathological clearance for excision of BCC in a tertiary care center and find the frequency of involvement of surgical margin with reference to anatomical site, with size and surgical technique. It was prospective open label interventional study conducted at Dermatology department of tertiary care hospital Rawalpindi Pakistan in lais on with histopathology department from January 2023 to April 2024. Total of thirty-six (n = 36) patients between age 45-80 years with basal cell carcinoma of 10-20mm on face were included following inclusion exclusion criteria by purposive sampling technique. Informed consent was taken. Surgical excision was performed and intraoperative frozen section histopathology clearance of tumor margin was taken from histopathologist on telephone. Surgical reconstruction was done. Final Histopathology report was reexamined on day 10th for margin and depth clearance. Descriptive statistics were calculated for age, gender, sun exposure, reconstructive technique, anatomical site, and tumor free margin report on frozen section analysis. Chi square test was employed for statistical significance of involvement of surgical margin with reference to anatomical site, size and decision on reconstructive surgical technique, p value of <0.05 was considered significant. Total of 36 patients of BCC were enrolled, males 12 (33.3%) and females were 24 (66.6%). Age ranged from 45 year to 80 year mean of 58.36 ±SD7.8. Size of BCC ranged from 10mm to 35mm mean of 25mm ±SD 0.63. Morphology was nodular 18 (50%), superficial spreading 11(30.6%), morphoeic 1 (2.8%) and ulcerative in 6(16.7%) cases. Intraoperative frozen section for histopathological margin clearance with 2-3 mm safety margin and surgical technique has p-value0.51, for anatomical site p value 0.24 and size p-0.84. Intraoperative frozen section (FS) histopathological clearance for BCC face with 2-3mm safety margin with reference to reconstructive technique, anatomical site and size of BCC were insignificant.

Keywords: basal cell carcinoma, tumor free amrgin, basal cell carcinoma and frozen section, safety margin

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Authors: Justine A. Lloyd

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Since the onset of the Syrian Civil War in 2011, the ancient city of Dura-Europos has faced widespread destruction and looting. The site is one of many places in the country the terrorist group ISIS has specifically targeted, allegedly due to its particular representations of Syrian history and culture. However, looted art and artifacts are the extremist group’s second largest source of income, only after oil. The protection of this site is important to both academics and the millions who have called Syria a home, as it aids in the nation’s sense of identity, reveals developments in the arts, and contributes to humanity’s collective history. At a time when Syria’s culture is being flattened, this sense of cultural expression is especially important to maintain. Creating an awareness of the magnitude of the issue at hand begins with an examination of the rich history of the ancient fortress city. Located on the western bank of the Euphrates River, Dura-Europos contains artifacts dating back to the Hellenistic, Parthian, and Roman periods. Though a great deal of the art and artifacts have remained safe in institutions such as the National Museum of Damascus and the Yale University Art Gallery, hundreds of looting pits and use of heavy machinery on the site has severely set back the investigative progress made by archaeologists over the last century, as well as the prospect of future excavation. Further research draws on the current destruction of the site by both ISIS and opportunists involved with the black market. Because Dura-Europos is located in a war stricken region, the acquisition of data and possibility of immediate action is particularly challenging. Resources gained from local reports, in addition to technology such as satellite imagery, however, have provided a firm starting point for the evaluation of the state of the site. The Syrian Ministry of Culture, UNESCO, and numerous Syrian and global organizations provide insight into the historic city’s past, present issues, and future plans to ensure that the cultural integrity of the site is upheld. Though over seventy percent of Dura-Europos has been completely decimated, this research challenges the notion that physically destroyed sites are lost forever. This paper assesses preventative measures that can take place to ensure the preservation of the site’s art and architecture, including examining possible solutions to the damage, such as digital reconstruction, replication, and distribution of information through exhibitions and other forms of publically accessible information. In order to investigate any possible retribution, research also includes the necessary information pertaining the global laws and regulations dealing with cultural heritage, as it directly affects the ways in which this situation can be dealt with. With the countless experts and citizens dedicated to the importance of cultural heritage, the prospect of honoring and valuing elements of Dura-Europos is possible—whether physically preserved or otherwise.

Keywords: antiquities law, archaeological sites, restitution, Syrian Civil War

Procedia PDF Downloads 147

Authors: Wai Yan Shih, Sau Man Wong, Wing Chung Chang, Wai Chi Chan

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Background: Late-life depression is associated with significant consequences. Although symptomatic reduction is achievable through pharmacological interventions, older adults are more vulnerable to the side effects than their younger counterparts. In addition, drugs do not address underlying cognitive dysfunctions such as rumination and reduced autobiographical memory specificity (AMS), both shown to be maladaptive coping styles that are associated with a poorer prognosis in depression. Considering how aging is accompanied by cognitive, psychological and physical changes, the interplay of these age-related factors may potentially aggravate and interfere with these depressive cognitive dysfunctions in late-life depression. Special care should, therefore, be drawn to ensure these cognitive dysfunctions are adequately addressed. Aim: This randomized controlled trial aims to examine the effect of mindfulness-based cognitive therapy (MBCT) on depressed older adults, and whether the potential benefits of MBCT are mediated by improvements in rumination and AMS. Method: Fifty-seven participants with an average age of 70 years old were recruited from multiple elderly centers and online mailing lists. Participants were assessed with: (1) Hamilton depression scale, (2) ruminative response scale, (3) autobiographical memory test, (4) mindful attention awareness scale, and (5) Montreal cognitive assessment. Eligible participants with mild to moderate depressive symptoms and normal cognitive functioning were randomly allocated to an 8-week MBCT group or active control group consisting of a low-intensity exercise program and health education. Post-intervention assessments were conducted after the 8-week program. Ethics approval was given by the Institutional Review Board of the University of Hong Kong/Hospital Authority. Results: Mixed-factorials ANOVAs demonstrated significant time x group interaction effects for depressive symptoms, AMS, and dispositional mindfulness. A marginally significant interaction effect was found for rumination. Simple effect analyses revealed a significant reduction in depressive symptoms for the both the MBCT group (mean difference = 7.1, p = .000), and control group (mean difference = 2.7, p = .023). However, only participants in the MBCT group demonstrated improvements in rumination, AMS, and dispositional mindfulness. Bootstrapping-based mediation analyses showed that the effect of MBCT in alleviating depressive symptoms was only mediated by the reduction in rumination. Conclusions: The findings support the use of MBCT as an effective intervention for depressed older adults, considering the improvements in depressive symptoms, rumination, AMS and dispositional mindfulness despite their age. Reduction in ruminative tendencies plays a major role in the cognitive mechanism of MBCT.

Keywords: mindfulness-based cognitive therapy, depression, older adults, rumination, autobiographical memory specificity

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Authors: Li Na Zhao, Arieh Warshel

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The design of enzymes is an extremely challenging task, and this is also true for metalloenzymes. In the case of naturally evolved enzymes, one may consider the active site residues as the musicians in the enzyme orchestra, while the metal can be considered as their concertmaster. Together they catalyze reactions as if they performed a masterpiece written by nature. The Lactonase can be thought as a member of the amidohydrolase family, with two concertmasters, Fe and Zn, at its active site. It catalyzes the quorum sensing signal- N-acyl homoserine lactones (AHLs or N-AHLs)- by hydrolyzing the lactone ring. This process, known as quorum quenching, provides a strategy in the treatment of infectious diseases without introducing selection pressure. However, the activity of lactonase is metal-dependent, and this dependence hampers the clinic usage. In our study, we use the empirical valence bond (EVB) approach to evaluate the catalytic contributions decomposing them to electrostatic and other components.

Keywords: enzyme redesign, empirical valence bond, lactonase, quorum quenching

Procedia PDF Downloads 231

Authors: David Maina Muthegethi

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The Victoria Basin is associated with gateway for migration to Southern part of Africa. Different communities migrated through the region including the Bantus and Nilotic communities that occupy present day Kenya and Tanzania. A distinct culture of dry-stone technology emerged around 15th century current era, a period associated with peopling of the western Kenya region. One of the biggest dry-stone walls enclosure is Thimlich Ohinga archaeological site. The site was constructed around fourteenth century current era. Architectural design was oval shaped stone structures that were around 4 meters and 2 meters in length and width respectively. The main subsistence strategies of the community that was crop faming, pastoralism, fishing, hunting and gathering. This paper attempts to examine gender dynamics of Thimlich Ohinga society. At that end, attempts are made to infer gender roles as manifested in archaeological record. Therefore, the study entails examination of material evidence excavated from the site. Also, ethnoarchaeological study of contemporary Luo community was undertaken in order to make inferences and analogies concerning gender roles of Thimlich Ohinga society. Overall, the study involved examination of cultural materials excavated from Thimlich Ohinga, extensive survey of the site and ethnography of Luo community. In total, an extensive survey and interviews of 20 households was undertaken in South Kanyamkango ward, Migori County in Western Kenya. The key findings point out that Thimlich Ohinga gender identities were expressed in material forms through architecture, usage of spaces, subsistence strategies, dietary patterns and household organization. Also, gender as social identity was dynamic and responsive to diversification of subsistence strategies and intensification of regional trade as documented in contemporary Luo community. The paper reiterates importance of ethnoarchaeological methods in reconstruction of past social organization as manifested in material record.

Keywords: ethnoarchaeological, gender, subsistence patterns, Thimlich Ohinga

Procedia PDF Downloads 59

Authors: Rahel Birhanu Kassaye, Ralf Otterpohl, Kumelachew Yeshitila

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Water resource management and land degradation are among the critical issues threatening the suitable livability of many cities in developing countries such as Ethiopia. Rapid expansion of urban areas and fast growing population has increased the pressure on water security. On the other hand, the large transformation of natural green cover and agricultural land loss to settlement and industrial activities such as quarrying is contributing to environmental concerns. Integrated water harvesting is considered to play a crucial role in terms of providing alternative water source to insure water security and helping to improve soil condition, agricultural productivity and regeneration of ecosystem. Moreover, it helps to control stormwater runoff, thus reducing flood risks and pollution, thereby improving the quality of receiving water bodies and the health of inhabitants. The aim of this research was to investigate the potential of applying integrated water harvesting approaches as a provision for water source and enabling land restoration in Jemo river catchment consisting of abandoned quarry site adjacent to a settlement area that is facing serious water shortage in western hilly part of Addis Ababa city, Ethiopia. The abandoned quarry site, apart from its contribution to the loss of aesthetics, has resulted in poor water infiltration and increase in stormwater runoff leading to land degradation and flooding in the downstream. Application of GIS and multi-criteria based analysis are used for the assessment of potential water harvesting technologies considering the technology features and site characteristics of the case study area. Biophysical parameters including precipitation, surrounding land use, surface gradient, soil characteristics and geological aspects are used as site characteristic indicators and water harvesting technologies including retention pond, check dam, agro-forestation employing contour trench system were considered for evaluation with technical and socio-economic factors used as parameters in the assessment. The assessment results indicate the different suitability potential among the analyzed water harvesting and restoration techniques with respect to the abandoned quarry site characteristics. Application of agro-forestation with contour trench system with the revegetation of indigenous plants is found to be the most suitable option for reclamation and restoration of the quarry site. Successful application of the selected technologies and strategies for water harvesting and restoration is considered to play a significant role to provide additional water source, maintain good water quality, increase agricultural productivity at urban peri-urban interface scale and improve biodiversity in the catchment. The results of the study provide guideline for decision makers and contribute to the integration of decentralized water harvesting and restoration techniques in the water management and planning of the case study area.

Keywords: abandoned quarry site, land reclamation and restoration, multi-criteria assessment, water harvesting

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Authors: Mohsen Sanayei, Jerzy Szpunar

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The present study investigates the effects of deformation strain and annealing temperature on the formation of twin boundaries, deformation and recrystallization texture evolution and grain boundary networks and connectivity. The resulting microstructures were characterized using Electron Backscatter Diffraction (EBSD) and X-Ray Diffraction (XRD) both immediately following small amount of deformation and after short time annealing at high temperature to correlate the micro and macro texture evolution of these alloys. Furthermore, this study showed that the process of grain boundary engineering, consisting cycles of deformation and annealing, is found to substantially reduce the mass and size of random boundaries and increase the proportion of low Coincidence Site Lattice (CSL) grain boundaries.

Keywords: coincidence site lattice, grain boundary engineering, electron backscatter diffraction, texture, x-ray diffraction

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Authors: Issa S. Oweis, Gary Gartenberg, Luma J. Oweis

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The site for a residential apartment building overlies an abandoned iron mine in granitic gneiss in northern New Jersey. The mine stope is about 137 m (450 long) and dipping over 344m (800 feet) at 450 to 500. As the building footprint straddles, the mine site needed remediation. The remediation scheme consisted of compaction grouting a minimum 10 m (30 ft.) depth of the mine stope in rock to establish a buttress for the hanging wall and allow support of the building foundation. The rock strength parameters (friction and cohesion) were established based on Hoek Geologic Strength Index (GSI). The derived strength parameters were used in the wedge analysis to simulate rock cave-in. It was concluded that a cave-in would be unlikely. Verification holes confirmed the effectiveness of grouting. Although post grouting micro gravity survey depicted a few anomalies, no anomalies were found to exist by further drilling and excavation.

Keywords: grout, stope, rock, properties

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Authors: Yuanyuan Liu, Yuanqing Wang, Di Li

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Carbon dioxide (CO2) emissions mitigation from road construction activities is one of the potential pathways to deal with climate change due to its higher use of materials, machinery energy consumption, and high quantity of vehicle and equipment fuels for transportation and on-site construction activities. Aiming to assess the environmental impact of the road infrastructure construction activities and to identify hotspots of emissions sources, this study developed a life-cycle CO2 emissions assessment framework covering three stages of material production, to-site and on-site transportation under the guidance of the principle of LCA ISO14040. Then streamlined inventory analysis on sub-processes of each stage was conducted based on the budget files from cases of highway projects in China. The calculation results were normalized into functional unit represented as ton per km per lane. Then a comparison between the amount of emissions from each stage, and sub-process was made to identify the major contributor in the whole highway lifecycle. In addition, the calculating results were used to be compared with results in other countries for understanding the level of CO2 emissions associated with Chinese road infrastructure in the world. The results showed that materials production stage produces the most of the CO2 emissions (for more than 80%), and the production of cement and steel accounts for large quantities of carbon emissions. Life cycle CO2 emissions of fuel and electric energy associated with to-site and on-site transportation vehicle and equipment are a minor component of total life cycle CO2 emissions from highway project construction activities. Bridges and tunnels are dominant large carbon contributor compared to the road segments. The life cycle CO2 emissions of road segment in highway project in China are slightly higher than the estimation results of highways in European countries and USA, about 1500 ton per km per lane. In particularly, the life cycle CO2 emissions of road pavement in majority cities all over the world are about 500 ton per km per lane. However, there is obvious difference between the cities when the estimation on life cycle CO2 emissions of highway projects included bridge and tunnel. The findings of the study could offer decision makers a more comprehensive reference to understand the contribution of road infrastructure to climate change, especially understand the contribution from road infrastructure construction activities in China. In addition, the identified hotspots of emissions sources provide the insights of how to reduce road carbon emissions for development of sustainable transportation.

Keywords: carbon dioxide emissions, construction activities, highway, life cycle assessment

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Authors: Yanhua Zhao, Yu Gou, Shu Feng, Dongdong Li, Chuanmin Tao

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The natural history of HBV infection could experience immune tolerant (IT), immune clearance (IC), HBeAg-negative inactive/quienscent carrier (ENQ), and HBeAg-negative hepatitis (ENH). As current biomarkers for discriminating these four phases have some weaknesses, additional serological indicators are needed. Hepatits B core-related antigen (HBcrAg) encoded with precore/core gene contains denatured HBeAg, HBV core antigen (HBcAg) and a 22KDa precore protein (p22cr), which was demonstrated to have a close association with natural history of hepatitis B infection, but no specific cutoff values and diagnostic parameters to evaluate the diagnostic efficacy. This study aimed to clarify the distribution of HBcrAg levels and evaluate its diagnostic performance during the natural history of infection from a Western Chinese perspective. 294 samples collected from treatment-naïve chronic hepatitis B (CHB) patients in different phases (IT=64; IC=72; ENQ=100, and ENH=58). We detected the HBcrAg values and analyzed the relationship between HBcrAg and HBV DNA. HBsAg and other clinical parameters were quantitatively tested. HBcrAg levels of four phases were 9.30 log U/mL, 8.80 log U/mL, 3.00 log U/mL, and 5.10 logU/mL, respectively (p < 0.0001). Receiver operating characteristic curve analysis demonstrated that the area under curves (AUCs) of HBcrAg and quantitative HBsAg at cutoff values of 9.25 log U/mL and 4.355 log IU/mL for distinguishing IT from IC phases were 0.704 and 0.694, with sensitivity 76.39% and 59.72%, specificity 53.13% and 79.69%, respectively. AUCs of HBcrAg and quantitative HBsAg at cutoff values of 4.15 log U/mlmL and 2.395 log IU/mlmL for discriminating between ENQ and ENH phases were 0.931 and 0.653, with sensitivity 87.93% and 84%, specificity 91.38% and 39%, respectively. Therefore, HBcrAg levels varied significantly among four natural phases of HBV infection. It had higher predictive performance than quantitative HBsAg for distinguishing between ENQ-patients and ENH-patients and similar performance with HBsAg for the discrimination between IT and IC phases, which indicated that HBcrAg could be a potential serological marker for CHB.

Keywords: chronic hepatitis B, hepatitis B core-related antigen, hepatitis B surface antigens, hepatitis B virus

Procedia PDF Downloads 387

Authors: Shirin jalili, Hadi Shirzad, Mahasti Modarresi, Samaneh Nabavi, Somayeh Khanjani

Abstract:

Identifying the source tissue of biological material found at crime scenes can be very informative in a number of cases. Despite their usefulness, current visual, catalytic, enzymatic, and immunologic tests for presumptive and confirmatory tissue identification are applicable only to a subset of samples, might suffer limitations such as low specificity, lack of sensitivity, and are substantially impacted by environmental insults. In addition their results are operator-dependent. Recently the possibility of discriminating body fluids using mRNA expression differences in tissues has been described but lack of long term stability of that Molecule and the need to normalize samples for each individual are limiting factors. The use of DNA should solve these issues because of its long term stability and specificity to each body fluid. Cells in the human body have a unique epigenome, which includes differences in DNA methylation in the promoter of genes. DNA methylation, which occurs at the 5′-position of the cytosine in CpG dinucleotides, has great potential for forensic identification of body fluids, because tissue-specific patterns of DNA methylation have been demonstrated, and DNA is less prone to degradation than proteins or RNA. Previous studies have reported several body fluid-specific DNA methylation markers.The presence or absence of a methyl group on the 5’ carbon of the cytosine pyridine ring in CpG dinucleotide regions called ‘CpG islands’ dictates whether the gene is expressed or silenced in the particular body fluid. Were described methylation patterns at tissue specific differentially methylated regions (tDMRs) to be stable and specific, making them excellent markers for tissue identification. The results demonstrate that methylation-based tissue identification is more than a proof-of-concept. The methodology holds promise as another viable forensic DNA analysis tool for characterization of biological materials.

Keywords: DNA methylation, forensic science, epigenome, tDMRs

Procedia PDF Downloads 409

Authors: Vanessa Roulet, Marc Turini, Juliane Hey, Stéphanie Bord-Romeu, Emilie Bonzom, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Vanessa Viotti, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

Abstract:

Background: Beckman Coulter, Inc. has recently developed fully automated assays for the detection of HBsAg on a new immunoassay platform. The objective of this European multicenter study was to evaluate the performance of the ACCESS HBsAg and ACCESS HBsAg Confirmatory assays† on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer. Methods: The clinical specificity of the ACCESS HBsAg and HBsAg Confirmatory assays was determined using HBsAg-negative samples from blood donors and hospitalized patients. The clinical sensitivity was determined using presumed HBsAg-positive samples. Sample HBsAg status was determined using a CE-marked HBsAg assay (Abbott ARCHITECT HBsAg Qualitative II, Roche Elecsys HBsAg II, or Abbott PRISM HBsAg assay) and a CE-marked HBsAg confirmatory assay (Abbott ARCHITECT HBsAg Qualitative II Confirmatory or Abbott PRISM HBsAg Confirmatory assay) according to manufacturer package inserts and pre-determined testing algorithms. False initial reactive rate was determined on fresh hospitalized patient samples. The sensitivity for the early detection of HBV infection was assessed internally on thirty (30) seroconversion panels. Results: Clinical specificity was 99.95% (95% CI, 99.86 – 99.99%) on 6047 blood donors and 99.71% (95%CI, 99.15 – 99.94%) on 1023 hospitalized patient samples. A total of six (6) samples were found false positive with the ACCESS HBsAg assay. None were confirmed for the presence of HBsAg with the ACCESS HBsAg Confirmatory assay. Clinical sensitivity on 455 HBsAg-positive samples was 100.00% (95% CI, 99.19 – 100.00%) for the ACCESS HBsAg assay alone and for the ACCESS HBsAg Confirmatory assay. The false initial reactive rate on 821 fresh hospitalized patient samples was 0.24% (95% CI, 0.03 – 0.87%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS HBsAg assay had equivalent sensitivity performances compared to the Abbott ARCHITECT HBsAg Qualitative II assay with an average bleed difference since first reactive bleed of 0.13. All bleeds found reactive in ACCESS HBsAg assay were confirmed in ACCESS HBsAg Confirmatory assay. Conclusion: The newly developed ACCESS HBsAg and ACCESS HBsAg Confirmatory assays from Beckman Coulter have demonstrated high clinical sensitivity and specificity, equivalent to currently marketed HBsAg assays, as well as a low false initial reactive rate. †Pending achievement of CE compliance; not yet available for in vitro diagnostic use. 2023-11317 Beckman Coulter and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. All other trademarks are the property of their respective owners.

Keywords: dxi 9000 access immunoassay analyzer, hbsag, hbv, hepatitis b surface antigen, hepatitis b virus, immunoassay

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Authors: Zeina Merabi, Arij Dao

Abstract:

The temporal order of information processing in the brain is an important code in many acoustic signals, including speech, music, and animal vocalizations. Despite its significance, surprisingly little is known about its underlying cellular mechanisms and network manifestations. In the songbird telencephalic nucleus HVC, a subset of neurons shows temporal combination sensitivity (TCS). These neurons show a high temporal specificity, responding differently to distinct patterns of spectral elements and their combinations. HVC neuron types include basal-ganglia-projecting HVCX, forebrain-projecting HVCRA, and interneurons (HVC¬INT), each exhibiting distinct cellular, electrophysiological and functional properties. In this work, we develop conductance-based neural network models connecting the different classes of HVC neurons via different wiring scenarios, aiming to explore possible neural mechanisms that orchestrate the combination sensitivity property exhibited by HVCX, as well as replicating in vivo firing patterns observed when TCS neurons are presented with various auditory stimuli. The ionic and synaptic currents for each class of neurons that are presented in our networks and are based on pharmacological studies, rendering our networks biologically plausible. We present for the first time several realistic scenarios in which the different types of HVC neurons can interact to produce this behavior. The different networks highlight neural mechanisms that could potentially help to explain some aspects of combination sensitivity, including 1) interplay between inhibitory interneurons’ activity and the post inhibitory firing of the HVCX neurons enabled by T-type Ca2+ and H currents, 2) temporal summation of synaptic inputs at the TCS site of opposing signals that are time-and frequency- dependent, and 3) reciprocal inhibitory and excitatory loops as a potent mechanism to encode information over many milliseconds. The result is a plausible network model characterizing auditory processing in HVC. Our next step is to test the predictions of the model.

Keywords: combination sensitivity, songbirds, neural networks, spatiotemporal integration

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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: 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

Abstract:

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: 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: Lintang Putra Sadewa, Ilham Prasetya Budhi

Abstract:

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

Abstract:

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: Amar Ranjan, Julieana Durai, Pranay Tanwar

Abstract:

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

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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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